Abstract

We report a young woman with the abrupt onset of infectious mononucleosis due to Epstein–Barr virus associated with a subsequent autoimmune form of diabetes mellitus and myasthenia gravis with anti–muscle-specific kinase (MuSK) antibodies. The simultaneous onset of these two autoimmune diseases preceded by a systemic viral illness supports a causal relationship between MuSK antibodies and myasthenia gravis and suggests the possibility of a viral trigger in some cases. Muscle Nerve, 2007

Abstract

Although half of dilated cardiomyopathy (DCM) cases have unknown causes, the remaining causes of DCM are extensive. In some cases, DCM starts with myocarditis, which is basically an inflammation of the heart muscle. Myocarditis can present in a variety of ways, from asymptomatic to cardiogenic shock. Myocarditis is associated with a wide range of infections, most commonly viral, bacterial, and parasitic. The main mechanism seems to be immune-mediated damage to the myocardium, which leads to a global or local ventricular dysfunction and DCM. Epstein-Barr virus is a rare cause of myocarditis. We could locate only 15 cases (9 adults and 6 children) reported in the medical literature in English. We report an adult patient who presented with signs and symptoms of heart failure most likely secondary to myocarditis caused by Epstein-Barr virus infection. The diagnosis was made by viral serology and a multidisciplinary approach.

Background
The association of oncogenic EBV with breast carcinoma (BC) is still in controversy.

Aim of work
Assess the association of EBV with BC in Egyptian women and find possible relationship between prognostic factors of BC and EBV detection.

Subjects and methods
Paraffin-embedded sections from 40 female patients with primary invasive BC; ductal (n = 32) and lobular (n = 8) and breast tissues from patients with fibrocystic disease (n = 20) as control were screened for presence of EBV by EBV nuclear antigen-1 (EBNA-1) immunostaining and by PCR for EBV-DNA.

Results
10 / 40 (25%) of the BC specimens stained positively for EBNA-1; EBNA-1 expression was restricted to a fraction 5%–60% of tumor epithelial cells. EBV-DNA was detected in 8 / 10 of BC specimens positive for EBNA-1. Control specimens were negative by both techniques. 7 / 8 (87.5%) of EBV-DNA positive tumors were associated with > 3 lymph nodes involvement.

Conclusion
EBV is associated with some invasive BC in Egyptian females and may play a role in their etiology.

Objective
To evaluate the effects of acute infection with Epstein–Barr virus (infectious mononucleosis, IM) on lipids and lipoproteins.

Methods
Fasting serum levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), triglycerides (TGs), apolipoproteins (apo) A-I, B, E, C-II, C-III and lipoprotein (a) [Lp(a)] were determined in patients with IM on diagnosis and 4 months after the resolution of febrile illness and in age- and sex-matched controls. Activities of cholesteryl-ester transfer protein (CETP), lipoprotein-associated phospholipase A₂ (Lp-PLA₂) and paraoxonase 1 (PON1) as well as levels of several cytokines were determined. LDL subclass analysis was performed with the Lipoprint LDL System.

Results
Twenty-nine patients (16 males, aged 24.3 ± 14.6 years) and 30 controls were included. TC, HDL-C, LDL-C, apoA-I, apoB, apoC-III and Lp(a) levels were lower at baseline whereas apoB/apoA-I ratio, TG levels and CETP activity were elevated compared with 4 months later. At baseline, higher levels in cytokines and the cholesterol concentration of small-dense LDL particles (sdLDL-C) were noticed, whereas LDL particle size was lower compared with follow-up. Activities of Lp-PLA₂ and PON1 were similar at baseline and 4 months later.

Four months after the resolution of IM levels of TGs, apoE, apoC-III, Lp(a), sdLDL-C and cytokines as well as LDL particle size, apoB/apoA-I ratio, CETP and Lp-PLA2 activities were similar to controls. PON1 activities both at baseline and 4 months later were lower in patients compared with controls.

Conclusions
IM is associated with atherogenic changes of lipids and lipoproteins that are partially restored 4 months after its resolution.

Background
The role of viruses in the context of sudden infant death in early childhood is still unclear, although there are many findings pointing to a viral infection possibly leading to death.

Objectives
To analyse the prevalence and viral loads of human herpesvirus-6 (HHV-6), Epstein-Barr virus (EBV), and cytomegalovirus (CMV), three viruses that have been previously detected in some cases of sudden death in infants, in tissues from sudden infant death syndrome (SIDS) patients and controls.

Materials and methods
Thirty-nine formalin-fixed paraffin-embedded tissue sections of eleven consecutive cases of SIDS, and thirty-nine formalin-fixed paraffin-embedded tissue sections of nine control cases were analysed by a specific quantitative real-time PCR for the detection of HHV-6, EBV, and CMV.

Results
The comparison of the whole viral DNA prevalence in cases and tissue sections between SIDS and controls showed a statistical significance (72.7% vs. 22.2%, p = 0.025; 41.1% vs. 10.3%, p = 0.001, respectively); in particular, we found a statistical significant difference for the EBV DNA prevalence among cases (p = 0.042) and tissues (p = 0.048), and a statistical significant difference for the HHV-6 DNA prevalence among cases (p = 0.017).

Conclusions
This is one of the first studies using quantitative real-time PCR for virus detection in cases of SIDS, and the results suggest that some herpesvirus infections, and particularly those caused by EBV and HHV-6 could be related with some cases of SIDS. Further studies will be necessary to understand the real significance of these findings in the context of SIDS.

In Graves' disease, the IgG class autoantibody against thyrotropin receptor (TRAb) is produced excessively and induces hyperthyroidism. Epstein-Barr virus (EBV) is one of the human herpesviruses that persists for life, mainly in B lymphocytes, and is occasionally reactivated. Therefore, EBV may affect the antibody production of B lymphocytes that would normally produce TRAb. The purpose of the present study was to evaluate the association of EBV reactivation with the etiology of Graves' disease. Serum levels of EBV antibodies and IgE were determined by ELISA. TRAb levels were determined by radioreceptor assay. We performed in-situ hybridization (ISH) of EBV-encoded small RNA (EBER)1 on the thyroid tissue of one of our patients. In Graves' disease patients with TRAb levels ≥10%, EA antibody levels, which indicate EBV reactivation, were moderately but significantly correlated with the levels of TRAb, and weakly but significantly correlated with IgE. EBER1-ISH revealed that one of our patients had EBV-infected lymphocytes infiltrating the thyroid gland. EBV reactivation may stimulate antibody-producing B lymphocytes predisposed to make TRAb, and this may contribute to or exacerbate the disease.

Objective
Increasing evidence supports a link between Epstein-Barr virus (EBV), a ubiquitous B-lymphotropic human herpesvirus, and common B-cell–related autoimmune diseases. We sought evidence of EBV infection in thymuses from patients with myasthenia gravis (MG), an autoimmune disease characterized by intrathymic B-cell activation.

Methods
Seventeen MG thymuses (6 follicular hyperplastic, 6 diffuse hyperplastic, 5 involuted) and 6 control thymuses were analyzed using in situ hybridization for EBV-encoded small RNAs (EBERs), immunohistochemistry for EBV latent and lytic proteins, and polymerase chain reaction for EBV DNA and mRNA.

Results
All 17 MG thymuses showed evidence of active EBV infection, whereas none of the control thymuses were infected. Cells expressing EBERs (12 of 17) and EBV latency proteins (EBNA2, LMP1, and LMP2A) (16 of 17) were detected in medullary infiltrates and in germinal centers. Cells expressing early (BFRF1, BMRF1) and late (p160, gp350/220) lytic phase EBV proteins were present in 16 MG thymuses. Latency (EBNA1, LMP2A) or lytic (BZLF1) transcripts (often both) were present in all MG thymuses, and EBV DNA (LMP1 gene) was detected in 13 MG thymuses. We also found CD8+ T cells, CD56 + CD3-natural killer cells, and BDCA-2+ plasmacytoid dendritic cells in immune infiltrates of MG thymuses, but not germinal centers, suggesting an attempt of the immune system to counteract EBV infection.

Interpretation
Dysregulated EBV infection in the pathological thymus appears common in MG and may contribute to the immunological alterations initiating and/or perpetuating the disease. ANN NEUROL 2010;67:726–738

Abstract

Bone marrow myelotoxicity is a major limitation of chemotherapy. While granulocyte colony stimulating factor (G-CSF) treatment is effective, alternative approaches to support hematopoietic recovery are sought. We previously found that a beta-glucan extract from maitake mushroom Grifola frondosa (MBG) enhanced colony forming unit-granulocyte monocyte (CFU-GM) activity of mouse bone marrow and human hematopoietic progenitor cells (HPC), stimulated G-CSF production and spared HPC from doxorubicin toxicity in vitro. This investigation assessed the effects of MBG on leukocyte recovery and granulocyte/monocyte function in vivo after dose intensive paclitaxel (Ptx) in a normal mouse. After a cumulative dose of Ptx (90–120 mg/kg) given to B6D2F1mice, daily oral MBG (4 or 6 mg/kg), intravenous G-CSF (80 µg/kg) or Ptx alone were compared for effects on the dynamics of leukocyte recovery in blood, CFU-GM activity in bone marrow and spleen, and granulocyte/monocyte production of reactive oxygen species (ROS). Leukocyte counts declined less in Ptx + MBG mice compared to Ptx-alone (p = 0.024) or Ptx + G-CSF treatment (p = 0.031). Lymphocyte levels were higher after Ptx + MBG but not Ptx + G-CSF treatment compared to Ptx alone (p < 0.01). MBG increased CFU-GM activity in bone marrow and spleen (p < 0.001, p = 0.002) 2 days after Ptx. After two additional days (Ptx post-day 4), MBG restored granulocyte/monocyte ROS response to normal levels compared to Ptx-alone and increased ROS response compared to Ptx-alone or Ptx + G-CSF (p < 0.01, both). The studies indicate that oral MBG promoted maturation of HPC to become functionally active myeloid cells and enhanced peripheral blood leukocyte recovery after chemotoxic bone marrow injury

Background: A polysaccharide extract from Grifola frondosa (Maitake extract) has shown immunomodulatory effects in preclinical studies and potentials for anticancer immunotherapy. However whether its oral administration in human is associated with measurable immunological change is unknown. Methods: In a phase I/II dose escalation trial, thirty postmenopausal breast cancer patients free of disease after initial treatment were enrolled sequentially into five cohorts of six patients each. Maitake extract was taken orally at 0.1, 0.5, 1.5, 3, or 5 mg/kg twice daily for three weeks. Peripheral blood was collected at day -7, 1 (prior to the first dosing), 7, 14, and 21 for ex vivo analyses. The primary endpoint was safety and tolerability. Results: No dose-limiting toxicity was encountered. Pharmacodynamic activities of the study agent did not follow a linear dose curve, as often seen with immunomodulatory agents. The following correlative endpoints showed significant changes compared to baseline. Quadratic dose curve analysis showed that a daily dosage of 2 mg/kg was associated with the greatest increases of CD3+CD25+ or CD4+CD25+ T cells in the peripheral blood (300% of baseline, p<0.001); a daily dosage of 6 mg/kg was associated with the most significant increases in intracellular IL-2 production by NK-T cells (237% of baseline, p=0.002); IL-10 production by T cells (360% baseline, p=0.002). Interferon gamma production by memory CD4+ T cells was attenuated to 27% of baseline at a daily dose of 7.4 mg/kg (p=0.002). Conclusions: Oral administration of a polysaccharide extract was associated with measurable changes in peripheral blood. The dose associated with the most significant changes varies by immunological parameter. The dose of 6 mg/kg is selected as the dose in future studies with clinical endpoints.

Abstract:

Introduction: Cancers are associated with excess cellular oxidative stress, and during cancer treatment the addition of drug-induced oxidative stress can limit the effectiveness of therapy and cause a number of side effects, such as fatigue, nausea, vomiting and diarrhea, as well as more serious adverse effects, including cardiomyopathy, peripheral neuropathy, hepatotoxicity and pulmonary fibrosis.

Method: Review of the pertinent literature on oxidative stress during cancer cytotoxic therapy and the use of Molecular Replacement methods to reduce adverse effects by replacement of damaged cellular molecules.

Discussion: Most of the adverse effects of cancer therapy are due to oxidative stress-mediated damage to normal tissues. For example, loss of efficiency in the electron transport chain caused by membrane peroxidation and reduction in coenzyme Q10 can occur during cytotoxic therapy using anthracyclines, alkylating agents, platinum coordination complexes, epipodophyllotoxins and camptothecins. Molecular Replacement and antioxidant administration mitigates the damage to normal tissues and reduces the adverse effects of cancer therapy without loss of therapeutic effect.

Summary: The acute and chronic adverse effects of cancer chemotherapy can be reduced by Molecular Replacement. Molecular Replacement of membrane lipids and enzymatic cofactors, such as coenzyme Q10, by administering nutritional supplements with antioxidants can prevent oxidative membrane damage and reductions of cofactors in normal tissues, respectively, restoring mitochondrial and other cellular functions and reducing chemotherapy adverse effects, such as cardiotoxicity, without significantly affecting therapeutic benefit. Recent clinical trials using cancer and non-cancer patients with chronic fatigue have shown the benefit of Molecular Replacement Therapy plus antioxidants in reducing the damage to mitochondrial membranes, restoring mitochondrial electron transport function, reducing fatigue and protecting cellular structures and enzymes from oxidative damage.

Abstract

Metastatic cancers are associated with cellular oxidative stress, and during cancer chemotherapy excess drug-induced oxidative stress can limit therapeutic effectiveness and cause a number of side effects, including fatigue, nausea, vomiting, diarrhea and more serious adverse effects, such as cardiomyopathy, peripheral neuropathy, hepatotoxicity and pulmonary fibrosis. We review here the hypothesis that the acute and chronic adverse effects of cancer chemotherapy can be reduced by molecular replacement of membrane lipids and enzymatic cofactors, such as coenzyme Q10. By administering nutritional supplements with replacement molecules and antioxidants, oxidative membrane damage and reductions of cofactors in normal tissues can be reversed, protecting and restoring mitochondrial and other cellular functions and reducing chemotherapy adverse effects. Recent clinical trials using cancer and non-cancer patients with chronic fatigue have shown the benefit of molecular replacement plus antioxidants in reducing the damage to mitochondrial membranes, restoring mitochondrial electron transport function, reducing fatigue and protecting cellular structures and enzymes from oxidative damage. Molecular replacement and antioxidant administration mitigates the damage to normal tissues, such as cardiac tissue, and reduces the adverse effects of cancer therapy without reduction in therapeutic results.

Abstract

Because most cancers are caused by dysregulation of as many as 500 different genes, agents that target multiple gene products are needed for prevention and treatment of cancer. Curcumin, a yellow coloring agent in turmeric, has been shown to interact with a wide variety of proteins and modify their expression and activity. These include inflammatory cytokines and enzymes, transcription factors, and gene products linked with cell survival, proliferation, invasion, and angiogenesis. Curcumin has been found to inhibit the proliferation of various tumor cells in culture, prevents carcinogen-induced cancers in rodents, and inhibits the growth of human tumors in xenotransplant or orthotransplant animal models either alone or in combination with chemotherapeutic agents or radiation. Several phase I and phase II clinical trials indicate that curcumin is quite safe and may exhibit therapeutic efficacy. These aspects of curcumin are discussed further in detail in this review.

Abstract

The cancer stem cell hypothesis asserts that malignancies arise in tissue stem and/or progenitor cells through the dysregulation or acquisition of self-renewal. In order to determine whether the dietary polyphenols, curcumin, and piperine are able to modulate the self-renewal of normal and malignant breast stem cells, we examined the effects of these compounds on mammosphere formation, expression of the breast stem cell marker aldehyde dehydrogenase (ALDH), and Wnt signaling. Mammosphere formation assays were performed after curcumin, piperine, and control treatment in unsorted normal breast epithelial cells and normal stem and early progenitor cells, selected by ALDH positivity. Wnt signaling was examined using a Topflash assay. Both curcumin and piperine inhibited mammosphere formation, serial passaging, and percent of ALDH+ cells by 50% at 5 μM and completely at 10 μM concentration in normal and malignant breast cells. There was no effect on cellular differentiation. Wnt signaling was inhibited by both curcumin and piperine by 50% at 5 μM and completely at 10 μM. Curcumin and piperine separately, and in combination, inhibit breast stem cell self-renewal but do not cause toxicity to differentiated cells. These compounds could be potential cancer preventive agents. Mammosphere formation assays may be a quantifiable biomarker to assess cancer preventive agent efficacy and Wnt signaling assessment can be a mechanistic biomarker for use in human clinical trials.

Abstract:

Curcumin, a natural component of the rhizome of curcuma longa has emerged as one of the most powerful chemopreventive and anticancer agents. Its biological effects range from antioxidant, anti-inflammatory to inhibition of angiogenesis and is also shown to possess specific antitumoral activity. The molecular mechanism of its varied cellular effects has been studied in some details and it has been shown to have multiple targets and interacting macromolecules within the cell. Curcumin has been shown to possess anti-angiogenic properties and the angioinhibitory effects of curcumin manifest due to down regulation of proangiogenic genes such as VEGF and angiopoitin and a decrease in migration and invasion of endothelial cells. One of the important factors implicated in chemoresistance and induced chemosensitivity is NFkB and curcumin has been shown to down regulate NFkB and inhibit IKB kinase thereby suppressing proliferation and inducing apoptosis. Cell lines that are resistant to certain apoptotic inducers and radiation become susceptible to apoptosis when treated in conjunction with curcumin. Besides this it can also act as a chemopreventive agent in cancers of colon, stomach and skin by suppressing colonic aberrant crypt foci formation and DNA adduct formation. This review focuses on the various aspects of curcumin as a potential drug for cancer treatment and its implications in a variety of biological and cellular processes vis-à-vis its mechanism of action.

ABSTRACT

Cadmium is a heavy metal that is found in the environment and enters our body through either dietary sources or cigarette smoke. Heavy metals, like cadmium tend to bioaccumulate and induce toxicities by interacting with organic compounds inside the cell. Recent studies have suggested that cadmium may play a role in breast cancer by activating the estrogen receptor (ER) (Antila1996, Stoica et al. 2000, Martin et al. 2003, Johnson et al. 2003). Data from our lab also suggest that acute exposure of cadmium promotes ER-dependent cell proliferation and induced ER target gene expression (i.e. cyclin D1, Cathepsin D, and c-myc). However the effects of chronic cadmium exposure are unclear and require further investigation. The current focus of this project is to understand how chronic exposure of cadmium affects breast cancer cell growth. To accomplish this, we have established a cadmium-breast cancer cell line (MCF-7 Cd) by maintaining parental ER+ MCF-7 cells in 10-7 M cadmium for over 3 months. By comparing the growth and gene expression patterns of MCF7-Cd and MCF7 cells, we hope to establish an understanding of how chronic cadmium exposure may contribute to the development of breast cancer.

Abstract

PURPOSE: Much debate has arisen about whether antioxidant supplementation alters the efficacy of cancer chemotherapy. Some have argued that antioxidants scavenge the reactive oxygen species integral to the activity of certain chemotherapy drugs, thereby diminishing treatment efficacy. Others suggest antioxidants may mitigate toxicity and thus allow for uninterrupted treatment schedules and a reduced need for lowering chemotherapy doses. The objective of this study is to systematically review the literature in order to compile results from randomized trials that evaluate concurrent use of antioxidants with chemotherapy.

DESIGN: MEDLINE, Cochrane, CinAhl, AMED, AltHealthWatch and EMBASE databases were searched. Only randomized, controlled clinical trials that reported survival and/or tumor response were included in the final tally. The literature searches were performed in duplicate following a standardized protocol. No meta-analysis was performed due to heterogeneity of tumor types and treatment protocols used in trials that met the inclusion criteria.

RESULTS: Of 845 articles considered, 19 trials met the inclusion criteria. Antioxidants evaluated were: glutathione (7), melatonin (4), vitamin A (2), an antioxidant mixture (2), vitamin C (1), N-acetylcysteine (1), vitamin E (1) and ellagic acid (1). Subjects of most studies had advanced or relapsed disease.

CONCLUSION: None of the trials reported evidence of significant decreases in efficacy from antioxidant supplementation during chemotherapy. Many of the studies indicated that antioxidant supplementation resulted in either increased survival times, increased tumor responses, or both, as well as fewer toxicities than controls; however, lack of adequate statistical power was a consistent limitation. Large, well-designed studies of antioxidant supplementation concurrent with chemotherapy are warranted.

Background

Although environmental lead exposure is associated with significant deficits in cognition, executive functions, social behaviors, and motor abilities, the neuroanatomical basis for these impairments remains poorly understood. In this study, we examined the relationship between childhood lead exposure and adult brain volume using magnetic resonance imaging (MRI). We also explored how volume changes correlate with historic neuropsychological assessments.

Methods and Findings

Volumetric analyses of whole brain MRI data revealed significant decreases in brain volume associated with childhood blood lead concentrations. Using conservative, minimum contiguous cluster size and statistical criteria (700 voxels, unadjusted p < 0.001), approximately 1.2% of the total gray matter was significantly and inversely associated with mean childhood blood lead concentration. The most affected regions included frontal gray matter, specifically the anterior cingulate cortex (ACC). Areas of lead-associated gray matter volume loss were much larger and more significant in men than women. We found that fine motor factor scores positively correlated with gray matter volume in the cerebellar hemispheres; adding blood lead concentrations as a variable to the model attenuated this correlation.

Conclusions

Childhood lead exposure is associated with region-specific reductions in adult gray matter volume. Affected regions include the portions of the prefrontal cortex and ACC responsible for executive functions, mood regulation, and decision-making. These neuroanatomical findings were more pronounced for males, suggesting that lead-related atrophic changes have a disparate impact across sexes. This analysis suggests that adverse cognitive and behavioral outcomes may be related to lead's effect on brain development producing persistent alterations in structure. Using a simple model, we found that blood lead concentration mediates brain volume and fine motor function.

Background: The treatment of pelvic malignancies (uterine, cervix, rectum, etc) often include radiation. Radiation-induced late side effects (longer than 3 months) are developed in 5–10% of those patients. We wanted to investigate the efficacy of hyperbaric oxygen therapy (HBOT) in the management of those patients. Methods: Thirteen women were evaluated. The primary cancer sources were: cervix (7), vagina (2), uterus (2), rectum (1) and bladder (1). All patients were treated with a full pelvic dose of radiotherapy. Eight patients also underwent post-radiation surgery (4 post-hysterectomy, 1 post-colectomy, 1 post-vaginectomy, 1 post-cystectomy and 1 post-exanteration). Eleven patients suffered from pelvic pain, 7 from chronic cystitis (including 2 with vesico-vaginal fistulas), 7 had chronic proctitis (including 2 with recto-vaginal fistulas), 3 had long-standing vaginal ulcers, and one presented with a long-standing open skin wound following surgery. All patients underwent imaging studies and biopsies to rule out active malignant disease, and all received HBOT 100% oxygen, at 2 absolute atmospheres, for 90 minutes (2ATA 90 min). Results: The mean patient age was 61 years (range 32–88). The mean time between completion of radiation therapy and onset of symptoms was 32 months (range 4–60). The patients received an average of 27 HBOTs (range 16–40). Twelve patients reported improvement in pelvic pain, bladder and bowel symptoms and decrease in vaginal discharge. One patient developed lung metastasis and another developed pelvic recurrence. No patient reported side effects associated with HBOT. Conclusion: HBOT appears to be safe and effective in the management of pelvic radiation-induced late side effects, such as soft tissue necrosis (skin and vagina), cystitis, proctitis and fistulas.

Therapeutic irradiation is commonly used to treat clinically localized prostate cancer. Although radiation therapy is effective for localized prostate cancer, the rectum is often affected because of its proximity to the treated tumour. The incidence of severe chronic radiation proctitis induced by pelvic irradiation is reported to be 2–5%. The symptoms of chronic radiation proctitis include diarrhoea, tenesmus, abdominal pain and rectal bleeding, and perforation, necrosis, stenosis and ulceration of the intestine may occur. The natural history of chronic radiation proctitis is unpredictable. While some patients with mild symptoms may experience remission, others suffer a progressively worsening course that can result in a life-threatening condition.

Chronic radiation proctitis is generally considered to be difficult to manage. Medical treatment with a low–residue diet and steroidal or nonsteroidal enemas has been disappointing. Laser therapy or electrocoagulation is often used to halt rectal bleeding. Although these attempts are initially successful in many patients, recurrence of bleeding is a common problem. Although faecal diversion is the most common operative procedure, this may not always control rectal bleeding and surgical resection of the involved segment may be necessary. Surgical complications are common, with a morbidity and mortality of 10–80%; thus an effective alternative treatment is required, particularly as medical treatment often fails. Hyperbaric oxygen therapy (HBOT) was recently reported to be safe and effective for the treatment of chronic radiation proctitis. We report the outcome of HBOT in four patients with chronic radiation proctitis.

Fasting mongrel dogs underwent hyperbaric oxygen treatment (HBOT), recombinant tissue plasminogen activator (rt-PA) treatment, and simultaneous HBOT and rt-PA treatment following prior experimental left anterior descending coronary artery occlusion for 2 hours. Thrombosis in and around a copper coil was recorded angiographically at regular intervals, and immediately prior to treatment conclusion. Controls (n = 10) were untreated. Group two animals (n = 10) were treated additionally with 90 minutes of HBOT at 2 atm absolute. Group three animals (n = 8) were treated additionally with 50 mg rt-PA over 90 minutes. Group four animals (n = 10) were treated additionally with simultaneous HBOT and rt-PA over 90 minutes. Myocardial injury was determined by a combination of triphenyltetrazolium chloride histochemical staining and by formazan dye extraction. Damage was measured as a percent of left ventricular cross-sectional area studied. HBOT alone restored 35.9% of oxidative enzyme activity (p greater than 0.001). Treatment with rt-PA alone restored 48.9% of enzyme activity (p greater than 0.001). The combination of HBOT and simultaneous rt-PA resulted in 96.9% restoration of oxidative enzyme activity versus the control group (p greater than 0.001)

Background: The relation between plasma vitamin C and risk of stroke remains unclear. Although clinical trials showed no significant benefit of vitamin C supplementation in reducing stroke risk, they were not able to examine the relation between plasma vitamin C concentrations and stroke risk in a general population.

Objective: The objective was to examine the relation between baseline plasma vitamin C concentrations and risk of incident stroke in a British population.

Design: A population-based prospective study was conducted in 20 649 men and women aged 40–79 y without prevalent stroke at baseline and participating in the European Prospective Investigation into Cancer–Norfolk prospective population study. The participants completed a health questionnaire and attended a clinic during 1993–1997 and were followed up for incident strokes through March 2005.

Results: Over 196 713 total person-years (average follow-up: 9.5 y), 448 incident strokes occurred. In a Cox proportional hazards model, persons in the top quartiles of baseline plasma vitamin C concentrations had a 42% lower risk (relative risk: 0.58; 95% CI: 0.43, 0.78) than did those in the bottom quartile, independently of age, sex, smoking, body mass index, systolic blood pressure, cholesterol, physical activity, prevalent diabetes and myocardial infarction, social class, alcohol consumption, and any supplement use. Similar results were obtained after exclusion of persons with illnesses, users of ascorbic acid–containing supplements, and persons with a history of early strokes during the initial 2 y of follow-up.

Conclusions: Plasma vitamin C concentrations may serve as a biological marker of lifestyle or other factors associated with reduced stroke risk and may be useful in identifying those at high risk of stroke.

Abstract

Objective  To test the ability of alpha-lipoic acid (LA) to attenuate the development of collagen-induced arthritis (CIA) in mice.
Methods  Mice were divided into three groups and treated with intraperitoneal administration of LA (10 or 100 mg/kg) or placebo. Clinical, histologic, and biochemical parameters were assessed. Human synovial fibroblasts and peripheral blood mononuclear cells were cocultured in various concentrations of LA to evaluate the effects on osteoclastogenesis.
Results  LA was associated with a dose-dependent reduction of CIA, as well as preventing bone erosion and destructive changes. Intracellular reactive oxygen species in lymphocytes obtained from inguinal lymph nodes, which was significantly higher in CIA than control mice, was significantly reduced in CIA by LA. The concentrations of TNF-a, IL-1β, and IL-6 in the paws, and synovial NF-κB binding, all of which were markedly higher in CIA than control mice, were reduced by treatment with LA. In addition, LA inhibited the formation of human osteoclasts in vitro.
Conclusion  Amelioration of joint disease by LA was associated with reduction in oxidative stress, as well as inhibition of inflammatory cytokine activation and NF-κB DNA binding activity. Moreover, LA inhibited bone destruction in vivo and osteoclastogenesis in vitro. Collectively, these results indicate that LA may be a new adjunctive therapy for rheumatoid arthritis.

Purpose

The purpose of this article is to review current evidence available for -lipoic acid (ALA) and its ability to improve symptoms of peripheral diabetic neuropathy (PDN).

Methods

This article searched MEDLINE from 1966 to November 2005 to identify clinical trials that supplemented ALA to individuals with type 1 or type 2 diabetes and positive sensory symptoms of PDN. Clinical trials to be included in this review met specific criteria of randomization, double masking, and placebo-controlled design.

Results

The search results produced 5 clinical trials that met the prerequisites for this review. ALA appears to improve neuropathic symptoms and deficits when administered via parenteral supplementation over a 3-week period. Oral treatment with ALA appears to have more conflicting data whether it improves sensory symptoms or just neuropathic deficits alone. An oral regimen of ALA and optimal length of treatment remains unclear. Both parenteral and up to a 2-year time period of oral supplementation of ALA appears to be safe without affecting glycemic control.

Conclusions

Based on these results, ALA should be considered as a treatment option for patients with PDN. When discussing supplementation with patients, it is important to discuss potential side effects; vitamin, mineral, and drug interactions; and current evidence available regarding efficacy.

Abstract:

The heavy metal mercury is ubiquitously distributed in the environment resulting in permanent low-level exposure in human populations. Mercury can be encountered in three main chemical forms (elemental, inorganic, and organic) which can affect the immune system in different ways. In this review, we describe the effects of these various forms of mercury exposure on immune cells in humans and animals. In genetically susceptible mice or rats, subtoxic doses of mercury induce the production of highly specific autoantibodies as well as a generalized activation of the immune system. We review studies performed in this model and discuss their implications for the role of environmental chemicals in human autoimmunity.

Abstract
Based on in vitro studies and short-term in vivo studies, all mercurials were for a long time considered as prototypic immunosuppressive substances. Recent studies have confirmed that organic mercurials such as methyl mercury (MeHg) and ethyl mercury (EtHg) are much more potent immunosuppressors than inorganic mercury (Hg). However, Hg interacts with the immune system in the presence of a susceptible genotype to cause immunostimulation, antinucleolar antibodies targeting fibrillarin, and systemic immune-complex (IC) deposits, a syndrome called Hg-induced autoimmunity (HgIA). Recent studies in mice with a susceptible genotype has revealed that the immunosuppressive effect of MeHg and EtHg will within 1–3 weeks be superseded by immunostimulation causing an HgIA-like syndrome. At equimolar doses of Hg, MeHg has the weakest immunostimulating, autoimmunogen, and IC-inducing effect, while the effect of thimerosal is similar to that of inorganic mercury. The immunosuppression is caused by the organic mercurials per se. Since they undergo rapid transformation to inorganic Hg, studies are being undertaken to delineate the importance of the organic substances per se and the newly formed inorganic Hg for induction of autoimmunity.

Lipoic acid (LA) is an antioxidant that suppresses and treats an animal model of multiple sclerosis (MS), experimental autoimmune encephalomyelitis. The purpose of this study was to determine the pharmacokinetics (PK), tolerability and effects on matrix metalloproteinase-9 (MMP-9) and soluble intercellular adhesion molecule-1 (sICAM-1) of oral LA in patients with MS. Thirty-seven MS subjects were randomly assigned to one of four groups: placebo, LA 600 mg twice a day, LA 1200 mg once a day and LA 1200 mg twice a day. Subjects took study capsules for 14 days. We found that subjects taking 1200 mg LA had substantially higher peak serum LA levels than those taking 600 mg and that peak levels varied considerably among subjects. We also found a significant negative correlation between peak serum LA levels and mean changes in serum MMP-9 levels (t=-0.263, P=0.04). There was a significant dose response relationship between LA and mean change in serum sICAM-1 levels (P=0.03). We conclude that oral LA is generally well tolerated and appears capable of reducing serum MMP-9 and sICAM-1 levels. LA may prove useful in treating MS by inhibiting MMP-9 activity and interfering with T-cell migration into the CNS.

Aims
Alpha-lipoic acid (LA) is a commonly used dietary supplement that exerts anti-oxidant and anti-inflammatory effects in vivo and in vitro. We investigated the mechanisms by which LA may confer protection in models of established atherosclerosis.

Main methods
Watanabe heritable hyperlipidemic (WHHL) rabbits were fed with high cholesterol chow for 6 weeks and then randomized to receive either high cholesterol diet alone or combined with LA (20 mg/kg/day) for 12 weeks. Vascular function was analyzed by myography. The effects of LA on T cell migration to chemokine gradients was assessed by Boyden chamber. NF-κB activation was determined by measuring translocation and electrophoresis migration shift assay (EMSA).

Key findings
LA decreased body weight by 15 ± 5% without alterations in lipid parameters. Magnetic Resonance Imaging (MRI) analysis demonstrated that LA reduced atherosclerotic plaques in the abdominal aorta, with morphological analysis revealing reduced lipid and inflammatory cell content. Consistent with its effect on atherosclerosis, LA improved vascular reactivity (decreased constriction to angiotensin II and increased relaxation to acetylcholine and insulin), inhibited NF-κB activation, and decreased oxidative stress and expression of key adhesion molecules in the vasculature. LA reduced T cell content in atherosclerotic plaque in conjunction with decreasing ICAM and CD62L (l-selectin) expression. These effects were confirmed by demonstration of a direct effect of LA in reducing T cell migration in response to CCL5 and SDF-1 and decreasing T cell adhesion to the endothelium by intra-vital microscopy.

Significance
The present findings offer a mechanistic insight into the therapeutic effects of LA on atherosclerosis.

Abstract
The anti-oxidant lipoic acid (LA) potently suppresses clinical and pathologic disease in the animal model of multiple sclerosis, experimental autoimmune encephalomyelitis, by inhibiting the migration of pathogenic T cells to the spinal cord. The mechanism by which this occurs is largely unknown. In this report we demonstrate that LA induces increases in cyclic AMP, a known immunosuppressant, in human T cells. The increase in cAMP is associated with increased adenylyl cyclase activity and is partially blocked by prostanoid receptor antagonists. We present evidence that LA also stimulates cAMP production in natural killer (NK) cells. This novel mechanism of action is highly relevant to the immunomodulatory effects of LA and provides further support for the study of LA as a therapeutic agent for multiple sclerosis and other autoimmune diseases.

Alpha-lipoic acid (LA), the metabolic antioxidant, was evaluated for its potential to protect against Cr⁶⁺-induced DNA damage. Potassium dichromate was administered to Swiss albino mice orally ad libitum at the doses of 5, 10 or 25 mg/kg body weight in drinking water to set DNA damage in cells, which was characterized in mouse peripheral blood mononuclear cells and bone marrow cells using single-cell gel electrophoresis and analyses of generated comets for Tail moment, Tail DNA and Tail length. DNA damage was dose dependent. Cytoprotection by LA was remarkable. LA (5, 10 and 25 mg/kg body weight intraperitoneally) in pre-, co- and post-toxicant administration schedule abrogated DNA damage substantially in both cell types. Protection by LA was also dose dependent. LA annulled DNA damage by Cr⁶⁺ in plasmid relaxation assay. A negligible DNA damage resulted during interaction of Cr⁶⁺ and LA. Compared to ascorbate, LA emerged as a better antioxidant and least DNA damaging. In conclusion, our study advocated an experimental therapeutic research potential in LA against Cr⁶⁺-induced DNA damage for reduction of occupational cancer risk in humans.

Abstract
The study investigates if alpha-lipoic acid is neuroprotective against chemotherapy induced neurotoxicity, if mitochondrial damage plays a critical role in toxic neurodegenerative cascade, and if neuroprotective effects of alpha-lipoic acid depend on mitochondria protection.
We used an in vitro model of chemotherapy induced peripheral neuropathy that closely mimic the in vivo condition by exposing primary cultures of dorsal root ganglion (DRG) sensory neurons to paclitaxel and cisplatin, two widely used and highly effective chemotherapeutic drugs. This approach allowed investigating the efficacy of alpha-lipoic acid in preventing axonal damage and apoptosis and the function and ultrastructural morphology of mitochondria after exposure to toxic agents and alpha-lipoic acid. Our results demonstrate that both cisplatin and paclitaxel cause early mitochondrial impairment with loss of membrane potential and induction of autophagic vacuoles in neurons. Alpha-lipoic acid exerts neuroprotective effects against chemotherapy induced neurotoxicity in sensory neurons: it rescues the mitochondrial toxicity and induces the expression of frataxin, an essential mitochondrial protein with anti-oxidant and chaperone properties. In conclusion mitochondrial toxicity is an early common event both in paclitaxel and cisplatin induced neurotoxicity. Alpha-lipoic acid protects sensory neurons through its anti-oxidant and mitochondrial regulatory functions, possibly inducing the expression of frataxin. These findings suggest that alpha-lipoic acid might reduce the risk of developing peripheral nerve toxicity in patients undergoing chemotherapy and encourage further confirmatory clinical trials.

a-Lipoic acid (α-LA) is a neuroprotective metabolic antioxidant that has been shown to cross the blood brain barrier. We tested whether a-LA is capable to prevent MOG35–55-induced experimental autoimmune encephalomyelitis (EAE), an established model of multiple sclerosis (MS). Daily oral administration of a-LA, starting at the time of immunization, significantly prevented EAE progression as compared to control mice. This was associated with a reduction of CNS infiltrating T cells and macrophages as well as decreased demyelination. We then tested a-LA in a therapeutic protocol aimed at suppressing EAE after its onset. Intraperitoneal (i.p.), but not oral, administration of a-LA significantly prevented disease progression when compared to vehicle-treated controls. Similarly, we observed significant reduction of demyelination and inflammatory infiltration. This clinical effect was not due to an impairment of MOG35–55 recognition by encephalitogenic T cells. In contrast, MOG-specific T cells showed a decreased production of IFNγ and IL-4, suggesting an immunosuppressive activity on both Th1 and Th2 cytokines. In addition, a-LA inhibited the proteolytic activity of MMP2 and MMP9 only at very high doses. Our data indicate that a-LA can effectively interfere with the autoimmune reaction associated with EAE through mechanisms other than its antioxidant activity and supports further studies on the use of a-LA as a potential therapy for MS.

α-Lipoic acid (LA) is a disulfide compound that is produced in small quantities in cells, and functions naturally as a co-enzyme in the pyruvate dehydrogenase and α-ketoglutarate dehydrogenase mitochondrial enzyme complexes. In pharmacological doses, LA is a multifunctional antioxidant. LA has been used in Germany for over 30 years for the treatment of diabetes-induced neuropathy. In patients with type 2 diabetes, recent studies have reported that intravenous (i.v.) infusion of LA increases insulin-mediated glucose disposal, whereas oral administration of LA has only marginal effects. If the limitations of oral therapy can be overcome, LA could emerge as a safe and effective adjunctive antidiabetic agent with insulin sensitizing activity.

The authors describe the long-term survival of a patient with pancreatic cancer without any toxic adverse effects. The treatment regimen includes the intravenous -lipoic acid and low-dose naltrexone (ALA-N) protocol and a healthy lifestyle program. The patient was told by a reputable university oncology center in October 2002 that there was little hope for his survival. Today, January 2006, however, he is back at work, free from symptoms, and without appreciable progression of his malignancy. The integrative protocol described in this article may have the possibility of extending the life of a patient who would be customarily considered to be terminal. The authors believe that life scientists will one day develop a cure for metastatic pancreatic cancer, perhaps via gene therapy or another biological platform. But until such protocols come to market, the ALA-N protocol should be studied and considered, given its lack of toxicity at levels reported. Several other patients are on this treatment protocol and appear to be doing well at this time.

The authors describe the long-term survival of a patient with pancreatic cancer without any toxic adverse effects. The treatment regimen includes the intravenous -lipoic acid and low-dose naltrexone (ALA-N) protocol and a healthy lifestyle program. The patient was told by a reputable university oncology center in October 2002 that there was little hope for his survival. Today, January 2006, however, he is back at work, free from symptoms, and without appreciable progression of his malignancy. The integrative protocol described in this article may have the possibility of extending the life of a patient who would be customarily considered to be terminal. The authors believe that life scientists will one day develop a cure for metastatic pancreatic cancer, perhaps via gene therapy or another biological platform. But until such protocols come to market, the ALA-N protocol should be studied and considered, given its lack of toxicity at levels reported. Several other patients are on this treatment protocol and appear to be doing well at this time.

Abstract
Reactive oxygen species are thought to be involved in a number of types of acute and chronic pathologic conditions in the brain and neural tissue. The metabolic antioxidant  a-lipoate (thioctic acid, 1, 2-dithiolane-3-pentanoic acid; 1, 2-dithiolane-3 valeric acid; and 6,8-dithiooctanoic acid) is a low molecular weight substance that is absorbed from the diet and crosses the blood–brain barrier.  a-Lipoate is taken up and reduced in cells and tissues to dihydrolipoate, which is also exported to the extracellular medium; hence, protection is afforded to both intracellular and extracellular environments. Both  a-lipoate and especially dihydrolipoate have been shown to be potent antioxidants, to regenerate through redox cycling other antioxidants like vitamin C and vitamin E, and to raise intracellular glutathione levels. Thus, it would seem an ideal substance in the treatment of oxidative brain and neural disorders involving free radical processes. Examination of current research reveals protective effects of these compounds in cerebral ischemia-reperfusion, excitotoxic amino acid brain injury, mitochondrial dysfunction, diabetes and diabetic neuropathy, inborn errors of metabolism, and other causes of acute or chronic damage to brain or neural tissue. Very few neuropharmacological intervention strategies are currently available for the treatment of stroke and numerous other brain disorders involving free radical injury. We propose that the various metabolic antioxidant properties of  a-lipoate relate to its possible therapeutic roles in a variety of brain and neuronal tissue pathologies: thiols are central to antioxidant defense in brain and other tissues. The most important thiol antioxidant, glutathione, cannot be directly administered, whereas  a-lipoic acid can. In vitro, animal, and preliminary human studies indicate that  a-lipoate may be effective in numerous neurodegenerative disorders.  

The antioxidant a-lipoic acid (LA) is a naturally occurring compound that has been shown to possess promising anticancer activity because of its ability to preferentially induce apoptosis and inhibit proliferation of cancer cells relative to normal cells. However, the molecular mechanisms underlying the apoptotic effect of LA are not well understood. We report here that LA induced reactive oxygen species (ROS) generation and a concomitant increase in apoptosis of human lung epithelial cancer H460 cells. Inhibition of ROS generation by ROS scavengers or by overexpression of antioxidant enzymes glutathione peroxidase and superoxide dismutase effectively inhibited LA-induced apoptosis, indicating the role of ROS, especially hydroperoxide and superoxide anion, in the apoptotic process. Apoptosis induced by LA was found to be mediated through the mitochondrial death pathway, which requires caspase-9 activation. Inhibition of caspase activity by the pan-caspase inhibitor (z-VAD-FMK) or caspase-9-specific inhibitor (z-LEHD-FMK) completely inhibited the apoptotic effect of LA. Likewise, the mitochondrial respiratory chain inhibitor rotenone potently inhibited the apoptotic and ROS-inducing effects of LA, supporting the role of mitochondrial ROS in LA-induced cell death. LA induced down-regulation of mitochondrial Bcl-2 protein through peroxide-dependent proteasomal degradation, and overexpression of the Bcl-2 protein prevented the apoptotic effect of LA. Together, our findings indicate a novel pro-oxidant role of LA in apoptosis induction and its regulation by Bcl-2, which may be exploited for the treatment of cancer and related apoptosis disorders.

Background and objectives: In spontaneously hypertensive rats (SHRs), excess endogenous aldehydes bind sulfhydryl groups of membrane proteins, altering membrane Ca²⁺ channels and increasing cytosolic free calcium and blood pressure. The thiol compound, N-acetyl cysteine, normalizes elevated blood pressure in SHRs by binding excess endogenous aldehydes and normalizing membrane Ca²⁺ channels and cytosolic free calcium. The aim of the present study was to investigate whether a dietary supplementation of an endogenous fatty acid, α-lipoic acid, another thiol compound that is known to increase tissue cysteine and glutathione, can lower blood pressure and normalize associated biochemical and histopathological changes in SHRs.

Methods and results: Starting at 12 weeks of age, animals were divided into three groups of six animals each. Animals in the Wistar-Kyoto (WKY) rat control group and the SHR control group were given a normal diet, and the SHR-lipoic acid group was given a diet supplemented with lipoic acid (500 mg/kg feed) for the next 9 weeks. After 9 weeks, systolic blood pressure, platelet [Ca²⁺]i, plasma insulin and liver, kidney and aortic aldehyde conjugates were significantly higher in SHR controls as compared with WKY rat controls and the SHR lipoic acid group. SHR controls also showed smooth muscle cell hyperplasia in the small arteries and arterioles of the kidneys.

Conclusions: Dietary α-lipoic acid supplementation in SHRs lowered the systolic blood pressure, cytosolic [Ca²⁺]i, blood glucose and insulin levels, and tissue aldehyde conjugates, and attenuated adverse renal vascular changes

ABSTRACT
Objective The aim of this study was to investigate the effect of alpha-lipoic acid (ALA) treatment on endothelium-dependent and -independent vasodilatation, assessed by forearm blood flow (FBF), in patients with type 2 diabetes mellitus.

Research design and methods A total of 30 subjects with type 2 diabetes were included in this randomized, controlled, double-blinded, parallel group study. FBF responses to intra-arterial acetylcholine (ACh) and glycerol trinitrate (GTN) were measured before and after 21 days of intravenous treatment with 600 mg alpha-lipoic acid or placebo.

Results FBF responses were comparable at baseline. After treatment, FBF reactivity to ACh and GTN was unchanged in subjects receiving placebo. By contrast, ALA treatment increased endothelium-dependent vasodilatation to ACh (P < 0·05) but not to GTN compared with baseline.

Conclusions Intravenous ALA treatment improves endothelium-dependent vasodilatation in patients with type 2 diabetes, in the absence of effects on forearm vasomotor function. If this salutary action translates into vascular risk reduction remains to be established.

Purpose

The purpose of this article is to review current evidence available for -lipoic acid (ALA) and its ability to improve symptoms of peripheral diabetic neuropathy (PDN).

Methods

This article searched MEDLINE from 1966 to November 2005 to identify clinical trials that supplemented ALA to individuals with type 1 or type 2 diabetes and positive sensory symptoms of PDN. Clinical trials to be included in this review met specific criteria of randomization, double masking, and placebo-controlled design.

Results

The search results produced 5 clinical trials that met the prerequisites for this review. ALA appears to improve neuropathic symptoms and deficits when administered via parenteral supplementation over a 3-week period. Oral treatment with ALA appears to have more conflicting data whether it improves sensory symptoms or just neuropathic deficits alone. An oral regimen of ALA and optimal length of treatment remains unclear. Both parenteral and up to a 2-year time period of oral supplementation of ALA appears to be safe without affecting glycemic control.

Conclusions

Based on these results, ALA should be considered as a treatment option for patients with PDN. When discussing supplementation with patients, it is important to discuss potential side effects; vitamin, mineral, and drug interactions; and current evidence available regarding efficacy.

Mitochondria produce reactive oxygen species that may contribute to vascular dysfunction. Alpha-lipoic acid and acetyl-L-carnitine reduce oxidative stress and improve mitochondrial function. In a double blind, crossover study, we examined the effects of combined alpha-lipoic acid /acetyl-L-carnitine treatment and placebo (eight weeks per treatment) on vasodilator function and blood pressure in 36 subjects with coronary artery disease. Active treatment increased brachial artery diameter by 2.3% (P=0.008), consistent with reduced arterial tone. Active treatment tended to decrease systolic blood pressure for the whole group (P=0.07) and had a significantly effect in the subgroup with blood pressure above the median (151±20 to 142±18 mmHg, P=0.03) and in the subgroup with the metabolic syndrome (139±21 to 130±18 mmHg, P=0.03). Thus, mitochondrial dysfunction may contribute to the regulation of blood pressure and vascular tone. Further studies are needed to confirm these findings and determine the clinical utility of alpha-lipoic acid/acetyl-L-carnitine as antihypertensive therapy.

Abstract

Aims: To determine the efficacy and safety of 600 mg of a-lipoic acid given i.v. over 3 weeks in diabetic patients with symptomatic polyneuropathy.
Methods: We searched the database of VIATRIS GmbH, Frankfurt, Germany, for clinical trials of
-lipoic acid according to the following prerequisites: randomized, double-masked, placebo-controlled, parallel-group trial using a-lipoic acid infusions of 600 mg i.v. per day for 3 Weeks, except for weekends, in diabetic patients with positive sensory symptoms of polyneuropathy which were scored by the Total Symptom Score (TSS) in the feet on a daily basis. Four trials (ALADIN I, ALADIN III, SYDNEY, NATHAN II) comprised n=1258 patients (a-lipoic acid: n=716; placebo: n=542) met these eligibility criteria and were included in a metaanalysis based on the intention-to-treat principle. Primary analysis involved a comparison of the differences in TSS from baseline to the end of i.v. treatment between the groups treated with a-lipoic acid or placebo. Secondary analyses included daily changes in TSS, responder rates (³50% improvement in TSS), individual TSS components, Neuropathy Impairment Score (NIS), NIS of the lower limbs (NIS-LL), individual NIS-LL components, and the rates of adverse events.
Results: After 3 weeks the relative difference in favour of a-lipoic acid vs placebo was 24.1 (13.5-33.4)% (geometric mean with 95% confidence interval) for TSS and 16.0 (5.7-25.2)% for NIS-LL. The responder rates were 52.7% in patients treated with a-lipoic acid and 36.9% in those on placebo (p<0.05). On a daily basis there was a continuous increase in the magnitude of TSS improvement in favour of a-lipoic acid vs placebo which was noted first after 8 days of treatment. Among the individual components of the TSS, pain, burning, and numbness decreased in favour of a-lipoic acid as compared with placebo, while among the NIS-LL components pin-prick and touch-pressure sensation as well as ankle reflexes were improved in favour of a-lipoic acid after 3 weeks. The rates of adverse events did not differ between the groups.
Conclusions: The results of this meta-analysis provide evidence that treatment with a-lipoic acid (600 mg/day i.v.) over 3 weeks is safe and significantly improves both neuropathic symptoms and deficits to a clinically meaningful degree in diabetic patients with symptomatic polyneuropathy.

The authors, in a previous article, described the long-term survival of a man with pancreatic cancer and metastases to the liver, treated with intravenous alpha-lipoic acid and oral low-dose naltrexone (ALA/N) without any adverse effects. He is alive and well 78 months after initial presentation. Three additional pancreatic cancer case studies are presented in this article. At the time of this writing, the first patient, GB, is alive and well 39 months after presenting with adenocarcinoma of the pancreas with metastases to the liver. The second patient, JK, who presented to the clinic with the same diagnosis was treated with the ALA/N protocol and after 5 months of therapy, PET scan demonstrated no evidence of disease. The third patient, RC, in addition to his pancreatic cancer with liver and retroperitoneal metastases, has a history of B-cell lymphoma and prostate adenocarcinoma. After 4 months of the ALA/N protocol his PET scan demonstrated no signs of cancer. In this article, the authors discuss the poly activity of ALA: as an agent that reduces oxidative stress, its ability to stabilize NF_kB, its ability to stimulate pro-oxidant apoptosic activity, and its discriminative ability to discourage the proliferation of malignant cells. In addition, the ability of lowdose naltrexone to modulate an endogenous immune response is discussed. This is the second article published on the ALA/N protocol and the authors believe the protocol warrants clinical trial.

The authors describe the long-term survival of a patient with pancreatic cancer without any toxic adverse effects. The treatment regimen includes the intravenous -lipoic acid and low-dose naltrexone (ALA-N) protocol and a healthy lifestyle program. The patient was told by a reputable university oncology center in October 2002 that there was little hope for his survival. Today, January 2006, however, he is back at work, free from symptoms, and without appreciable progression of his malignancy. The integrative protocol described in this article may have the possibility of extending the life of a patient who would be customarily considered to be terminal. The authors believe that life scientists will one day develop a cure for metastatic pancreatic cancer, perhaps via gene therapy or another biological platform. But until such protocols come to market, the ALA-N protocol should be studied and considered, given its lack of toxicity at levels reported. Several other patients are on this treatment protocol and appear to be doing well at this time.

Abstract
Glutathione is an abundant intracellular thiol antioxidant whose levels are reduced both in Parkinson's disease itself and in a widely used animal model of the disorder, systemic MPTP administration. Previous in vitro work from our laboratory has suggested that glutathione depletion may be directly responsible for mitochondrial dysfunction, which ultimately leads to dopaminergic cell death associated with the disease. Here, we demonstrate the ability of gamma-glutamylcysteine ethyl ester, a lipid permeable derivative of the major substrate for scavenger glutathione synthesis, to counteract glutathione loss and neurodegeneration associated with in vitro and in vivo administration of MPTP or its derivatives. This data suggests that prevention of glutathione depletion is a likely therapeutic target for the disease.

Abstract

Parkinson's disease (PD) is associated with loss of total glutathione (GSH) which may contribute to progressive cell death. Peripheral GSH administration has been used clinically with reported benefits. Despite this, there is little specific information to characterize its cellular uptake or clearance, brain elevation with peripheral delivery or neuroprotective efficacy in PD models. The current study was carried out to provide this information using in vitro and in vivo approaches. In rat mesencephalic culture, the monoethyl ester of GSH (GEE), but not GSH (1–10 mM, 24 h) produced a dose-dependent elevation in GSH. The half-life for clearance was 10.14 h and was not different in cells depleted of GSH prior to loading. Elevation of GSH with GEE protected neurons from oxidative stress with H2O2 or metabolic stress with the complex I and II inhibitors MPP⁺ and malonate, respectively. To determine if peripheral administration of GEE could elevate brain GSH levels, rats were administered 0.1–50 mg/kg/day GEE via osmotic minipump either subcutaneously (sc) or via a cannula placed into the left cerebral ventricle (icv) for 28 days. Only central delivery of GEE resulted in significant elevations of brain GSH. Elevation of brain GSH by icv infusion of GEE was examined for its neuroprotective effects against chronic central delivery of MPP^+. Infusion of 0.142 mg/kg/day MPP⁺ for 28 days caused a selective ipsilateral loss of striatal dopamine. Co-infusion of MPP⁺ with 10 mg/kg/day GEE significantly protected against striatal dopamine loss. These findings show that the ethyl ester of GSH but not GSH per se can elevate intracellular GSH, that brain elevation of GSH requires central delivery of the ethyl ester and that this elevation provides neuroprotection against oxidative stress or chronic mitochondrial impairment.

Abstract

Parkinson's disease (PD) is characterized by early glutathione depletion in the substantia nigra (SN). Among its various functions in the cell, glutathione acts as a substrate for the mitochondrial enzyme glutaredoxin 2 (Grx2). Grx2 is involved in glutathionylation of protein cysteine sulfhydryl residues in the mitochondria. Although monothiol glutathione–dependent oxidoreductases (Grxs) have previously been demonstrated to be involved in iron-sulfur (Fe-S) center biogenesis, including that in yeast, here we report data suggesting the involvement of mitochondrial Grx2, a dithiol Grx, in iron-sulfur biogenesis in a mammalian dopaminergic cell line. Given that mitochondrial dysfunction and increased cellular iron levels are two important hallmarks of PD, this suggests a novel potential mechanism by which glutathione depletion may affect these processes in dopaminergic neurons. We report that depletion of glutathione as substrate results in a dose-dependent Grx2 inhibition and decreased iron incorporation into a mitochondrial complex I (CI) and aconitase (m-aconitase). Mitochondrial Grx2 inhibition through siRNA results in a corresponding decrease in CI and m-aconitase activities. It also results in significant increases in iron-regulatory protein (IRP) binding, likely as a consequence of conversion of Fe-S–containing cellular aconitase to its non–Fe-S–containing IRP1 form. This is accompanied by increased transferrin receptor, decreased ferritin, and subsequent increases in mitochondrial iron levels. This suggests that glutathione depletion may affect important pathologic cellular events associated with PD through its effects on Grx2 activity and mitochondrial Fe-S biogenesis.

Introduction
Cadmium induces hypertension in animal models. Epidemiologic studies of cadmium exposure and hypertension, however, have been inconsistent.

Objective
We aimed to investigate the association of blood and urine cadmium with blood pressure levels and with the prevalence of hypertension in U.S. adults who participated in the 1999–2004 National Health and Nutrition Examination Survey (NHANES).

Methods
We studied participants ≥ 20 years of age with determinations of cadmium in blood (n = 10,991) and urine (n = 3,496). Blood and urine cadmium were measured by atomic absorption spectrometry and inductively coupled plasma–mass spectrometry, respectively. Systolic and diastolic blood pressure levels were measured using a standardized protocol.

Results
The geometric means of blood and urine cadmium were 3.77 nmol/L and 2.46 nmol/L, respectively. After multivariable adjustment, the average differences in systolic and diastolic blood pressure comparing participants in the 90th vs. 10th percentile of the blood cadmium distribution were 1.36 mmHg [95% confidence interval (CI), −0.28 to 3.00] and 1.68 mmHg (95% CI, 0.57–2.78), respectively. The corresponding differences were 2.35 mmHg and 3.27 mmHg among never smokers, 1.69 mmHg and 1.55 mmHg among former smokers, and 0.02 mmHg and 0.69 mmHg among current smokers. No association was observed for urine cadmium with blood pressure levels, or for blood and urine cadmium with the prevalence of hypertension.

Conclusions
Cadmium levels in blood, but not in urine, were associated with a modest elevation in blood pressure levels. The association was stronger among never smokers, intermediate among former smokers, and small or null among current smokers. Our findings add to the concern of renal and cardiovascular cadmium toxicity at chronic low levels of exposure in the general population.

The activity response of the antioxidant enzymes glutathione peroxidase (GPx), glutathione reductase (GR) and the contents of thiobarbituric reactive substances (TBARS) were investigated in rats exposed to lead. The enzyme activities were determined in the liver, kidney and heart of male and female rats which were received 100 mg and 1000 mg of lead acetate per liter water for 18 weeks. The statistical analyses indicated the differences related to the organs and to the sex of animals. Administration of lead evoked decrease of GPx activity in the kidney of both male and female rats. On the contrary, GPx activity increased in the heart of female rats, while in the male rats the higher dose of lead evoked a decrease in activity. In the kidneys of male rats and in the heart of female rats thiobarbituric acid reactive substances (TBARS), an indicators of oxidative stress, significantly increased in rats which were given the high lead dose. Most likely the observed changes could be a compensatory response to different lead accumulation in the male and female organs and also the possible distinct mechanisms in ROS elimination.

Abstract
Prenatal and early-life exposure to lead is hypothesized to have a range of adverse effects on childhood health. Drawing on data collected from a population-based prospective cohort study of a highly exposed town and a low exposed town in Kosovo, Yugoslavia we assessed whether elevated maternal blood lead (BPb) concentrations during pregnancy were associated with reduced childhood measures of attained height and BMI or growth rate, and whether the associations, if any, were mediated by maternal thyroid hormone concentration at mid-pregnancy. There was no association between blood lead levels and height or BMI in either town. However, increased maternal thyroid hormone was unexpectedly associated with reduced attained childhood height, and growth rate of height from 6.5 to 10 years, in the low-exposure town. We examine potential reasons for this unexpected inverse association.

Abstract
In order to investigate the toxic effects of lead during occupational exposure to this metal and the antidotal efficacy of ascorbic acid directed against these toxic effects, various artisans in Abeokuta, Nigeria, who have been shown to be occupationally exposed to lead were supplemented daily with 500 mg ascorbic acid for 2 weeks. Ca²+-Mg²+-ATPase activity in erythrocyte membrane, as well as calcium and magnesium concentrations in plasma, erythrocytes, erythrocyte membrane and urine of the artisans were determined before and after ascorbic acid supplementation. The 2-week ascorbic acid administration resulted in the reversal of lead-induced accumulation of calcium and magnesium in the erythrocyte membranes of the artisans. Ascorbic acid also reversed lead-induced inhibition of erythrocyte membrane Ca²+-Mg²+-ATPase. Urinary excretion of calcium and magnesium was not affected by ascorbic acid. There may be some scope in introducing ascorbic acid as an intervention strategy for the prevention and therapy of lead intoxication, especially in cases where the subjects cannot be removed from the source of lead exposure.

Background:
DNA methylation is an epigenetic mark which regulates gene expression and can be modified by environmental factors, possibly including toxic metals. Although their function is unclear, DNA contains large numbers of interdispersed repetitive elements (ALU and LINE-1) that are normally heavily methylated. Global methylation can be measured within these elements and changes in global methylation have been associated with diseases such as cancer and cardiovascular disease. Changes in global methylation may be representative of the effects of environmental factors on epigenetic marks and may explain observed phenomena such as programming and the latencies between exposure and disease onset.

Methods:
We measured global methylation on a subset of 783 Normative Aging Study subjects (all male) who had archived blood DNA samples. We measured patella and tibia lead levels by X-Ray fluorescence. Blood lead levels were measured by atomic absorption spectrometry. We determined global DNA methylation averages within CpG repeats of LINE-1 and ALU elements using PCR-Pyrosequencing on bisulfite-treated DNA. We constructed a series of multiple linear regression models using methylation with ALU or LINE-1 as the dependent variable and bone lead (tibia or patella in separate models) as the primary exposure markers. All models were adjusted for age at DNA collection, pack-years of smoking, education and blood Pb at time of DNA collection as covariates.

Results:
Overall mean (SD) values for global methylation as measured by ALU and LINE-1 were as follows: ALU-26.3% STD(1.1%); LINE-1- 76.8% STD(1.8%), respectively. Mean (SD) age was 72.9 (7.6) years. 32% of subjects had more than a high school education. Mean (SD) pack-years of smoking was 23.5 STD(26). In both the bivariate and full model patella lead levels were inversely associated with ALU (standardized Beta = -0.1; P = 0.03) but not LINE-1; standardized Beta = -0.04; P = 0.4). Results were unchanged if blood Pb was dropped from the model. Among the covariates only age was associated with global methylation (standardized Beta = -0.1; P = 0.01) Tibia lead and blood lead did not predict global methylation for either ALU or LINE-1.

Conclusion:
Patella lead levels inversely predict reduced global DNA methylation within ALU elements. We believe this is the first report in humans of an association between chronic lead exposure and DNA methylation. This finding may have implications for lead's mechanism of action in producing adverse health effects. Changes in DNA methylation could be a mechanism for long term programming effects as well. Further study is needed to confirm these findings and to determine whether DNA methylation marks (global and gene specific) are useful biomarkers of lead toxicity in epidemiologic studies.

Introduction:
The WHO Environmental Burden of Disease (EBD) methodology was used to assess burden of disease from lead exposure in Ireland. The aim was to produce a national baseline assessment and to evaluate whether intervention was required.

Methods:
The outcomes used for children were IQ loss and resulting rates of mild mental retardation (MMR), rates of anaemia, and gastrointestinal symptoms were calculated for children. For adults, raised blood pressure as an indicator for risk of cardiovascular disease was used. Mean blood lead levels from a survey of a rural population living beside an old mining site were used and assumed to be representative of urban exposures (Adults = 3.0μg/dL, n=341; pre-school children= 2.7μg/dL, n=84). Communities beside mining sites have been found to have similar levels of exposure to urban populations. Urban populations are assumed to have a greater proportion of their exposure from transport sources so estimates were constructed with and without the use of a correction factor to account for annual reductions since the ban on lead in gasoline in 2000. Assuming a normal distribution of the log-transformed means, the proportion of the population at each blood Pb interval was estimated, and rates of disease for each health outcome were mapped to each interval, based on the WHO dose-response estimates.

Results:
The attributable incidence rates of MMR in children was low (0.25/1000), as were rates of gastrointestinal symptoms (0.06/1000) and anaemia (0.04/1000). The Attributable Fractions for cerebrovascular and hypertensive disease in men was 1% and 0% for ischaemic heart disease and other heart disease in men. The Attributable Fraction for all cardiovascular outcomes in women was 0%.

Discussion & Conclusions:
The study population used is not representative of the Irish population. Populations adjacent to mining sites do not experience exposures comparable to those observed in communities beside other point sources, such as smelters. The survey population may be more representative of rural exposures, making the current assessment an underestimate. Analysis and comparison with similar urban and rural populations will be conducted to determine whether the current assessment should be assumed as a low, medium or high estimate. In countries with long-standing legislative controls on the use of lead, exposure of the general population will be low. Locally high levels due to point sources and exposure of children living in degraded housing are likely to be more important. Targeted surveillance of identified 'at risk' groups as recommended by the CDC in the US may be more practical for informing public health than the wider EBD assessment. The study highlighted the difficulty in conducting epidemiology in the absence of any biomonitoring in Ireland.

Abstract
Lead remains one of the most hazardous toxins in our environment. Because the toxic effects of lead are most prominent during early development, it is important to develop a suitable biomarker for lead exposure during the pre- and neonatal periods. In the present study, the spatial distribution of lead was measured in the enamel and dentine of ten human primary teeth using laser ablation-inductively coupled plasma-mass spectrometry. The neonatal line, visualized using confocal laser scanning microscopy, was used to demarcate the pre- and postnatal regions of the sample teeth. Lead levels in pre- and postnatally formed enamel and dentine were compared to blood-lead levels measured at birth and one year of age for four of these participants. Mean dentine-lead levels ranged from 0.17 ± 0.02 to 5.60 ± 1.79 μg/g, and mean enamel-lead levels ranged from 0.04 ± 0.01 to 1.47 ± 0.20 μg/g. The results of this preliminary study showed that the spatial distribution of lead in dentine reflected the blood-lead levels. The present study demonstrates a methodology where the spatial distribution of lead in the dentine of human primary teeth may be used to obtain temporal information of environmental lead exposure during the pre- and neonatal periods.

Abstract
The purpose of the study was to assess the neurocognitive status of 6-month-old infants whose mothers were exposed to low but varying amounts of lead during pregnancy. Lead levels in the cord blood were used to assess environmental exposure and the Fagan Test of Infant Intelligence (FTII) assessed visual recognition memory (VRM). The cohort consisted of 452 infants of mothers who gave birth to babies at 33–42 weeks of gestation between January 2001 and March 2003. The overall mean lead level in the cord blood was 1.42 μg/dl (95% CI: 1.35–1.48). We found that VRM scores in 6 month olds were inversely related to lead cord blood levels (Spearman correlation coefficient −0.16, p=0.007). The infants scored lower by 1.5 points with an increase by one unit (1 μg/dl) of lead concentration in cord blood. In the lower exposed infants (1.67 μg/dl) the mean Fagan score was 61.0 (95% CI: 60.3–61.7) and that in the higher exposed group (>1.67 μg/dl) was 58.4 (95% CI: 57.3–59.7). The difference of 2.5 points was significant at the p=0.0005 level. The estimated risk of scoring the high-risk group of developmental delay (FTII classification 3) due to higher lead blood levels was two-fold greater (OR=2.33, 95% CI: 1.32–4.11) than for lower lead blood levels after adjusting for potential confounders (gestational age, gender of the child and maternal education). As the risk of the deficit in VRM score (Fagan group 3) in exposed infants attributable to Pb prenatal exposure was about 50%, a large portion of cases with developmental delay could be prevented by reducing maternal blood lead level below 1.67 μg/dl. Although the negative predictive value of the chosen screening criterion (above 1.67 μg/dl) was relatively high (89%) its positive predictive value was too low (22%), so that the screening program based on the chosen cord blood lead criterion was recommended.

Abstract
Parameters of semen quality, seminal plasma indicators of secretory function of the prostate and seminal vesicles, sex hormones in serum, and biomarkers of lead, cadmium, copper, zinc, and selenium body burden were measured in 240 Croatian men 19–52 years of age. The subjects had no occupational exposure to metals and no known other reasons suspected of influencing male reproductive function or metal metabolism. After adjusting for age, smoking, alcohol, blood cadmium, and serum copper, zinc, and selenium by multiple regression, significant (P<0.05) associations of blood lead (BPb), δ-aminolevulinic acid dehydratase (ALAD), and/or erythrocyte protoporphyrin (EP) with reproductive parameters indicated a lead-related increase in immature sperm concentration, in percentages of pathologic sperm, wide sperm, round sperm, and short sperm, in serum levels of testosterone and estradiol, and a decrease in seminal plasma zinc and in serum prolactin. These reproductive effects were observed at low-level lead exposure (BPb median 49 μg/L, range 11–149 μg/L in the 240 subjects) common for general populations worldwide. The observed significant synergistic effect of BPb and blood cadmium on increasing serum testosterone, and additive effect of a decrease in serum selenium on increasing serum testosterone, may have implications on the initiation and development of prostate cancer because testosterone augments the progress of prostate cancer in its early stages.

OBJECTIVES. The purpose of this work was to assess the long-term impact of childhood lead exposure on the neurosubstrate of language function and brain organization.

METHODS. Young adults from the Cincinnati Lead Study were recruited to undergo functional magnetic resonance image scanning while performing a verb generation task. These subjects have been followed from birth through early childhood with extensive documentation of lead exposure, neuropsychology, and behavior. Forty-two subjects provided useful imaging data. The locale, strength, and the correlation between brain language activation and childhood blood lead concentration were studied.

RESULTS. After adjusting for potential confounders, the activation in left frontal cortex, adjacent to Broca's area, and left middle temporal gyrus, including Wernicke's area, were found to be significantly associated with diminished activation in subjects with higher mean childhood blood lead levels, whereas the compensatory activation in the right hemisphere homolog of Wernicke's area was enhanced in subjects with higher blood lead levels.

CONCLUSION. This study indicates that childhood lead exposure has a significant and persistent impact on brain reorganization associated with language function.

Objective:
Accumulating evidence suggests that nonoccupational exposures to lead may increase the risk for cardiovascular and neurologic outcomes such as hypertension and accelerated decline in cognition. Chronic low-level exposures, as measured by bone lead levels, are likely at least as important as acute exposures, as measured in blood lead. Elevated levels of plasma homocysteine (Hcy) are associated with cognitive decline and dementia, as well as cardiovascular disease.

Materials and Methods:
To examine lead's potential contribution to Hcy-mediated pathways in the development of these conditions, we evaluated bone and lead levels in association with plasma Hcy levels assessed up to 3 times over 6 years in a sample of 900 older men (mean age: 69 years) living in the Boston area. Analyses were adjusted for age, education, smoking, alcohol consumption, intake of vitamins B6 and B12, folate intake, and waist circumference.

Results:
Higher levels of lead independently predicted significantly higher levels of plasma Hcy. These findings held whether lead was measured in blood, cortical bone (tibia), or trabecular bone (patella). An interquartile range increment in blood lead (3 μg/dL) was associated with plasma Hcy levels that were 0.74 nmol/L higher (95% CI: 0.48-1.00; P<0.001). This increase in Hcy was similar to the increase in serum Hcy we observed for study participants who were about 9 years apart in age. Results pertaining to bone lead were similar; for example, an interquartile range in tibia lead (14 μg/dL) was associated with plasma Hcy levels that were 0.35 nmol/L higher (95% CI: 0.11-0.60; P = 0.005).

Conclusions:
The single previously published study of lead exposure and Hcy found a significant association with blood lead, but not bone lead. Using repeated measurements of Hcy, our results complement and expand these prior findings, suggesting that lead may exert its neuro- and vascular toxicity, both acutely and chronically, by elevating Hcy.

Background
A few recent studies have demonstrated heightened hypothalamic–pituitary–adrenal (HPA) axis reactivity to acute stress in animals exposed to heavy metal contaminants, particularly lead. However, Pb-induced dysregulation of the HPA axis has not yet been studied in humans.

Objective
In this study, we examined children’s cortisol response to acute stress (the glucocorticoid product of HPA activation) in relation to low-level prenatal and postnatal Pb exposure.

Methods
Children’s prenatal blood Pb levels were determined from cord blood specimens, and postnatal lead levels were abstracted from pediatrician and state records. Children’s adrenocortical responses to an acute stressor were measured using assays of salivary cortisol before and after administration of a standard cold pressor task.

Results
Pb exposure was not associated with initial salivary cortisol levels. After an acute stressor, however, increasing prenatal and postnatal blood Pb levels were independently associated with significantly heightened salivary cortisol responses.

Conclusions
Our results suggest that relatively low prenatal and postnatal blood lead levels—notably those below the 10 μg/dL blood lead level identified by the Centers for Disease Control and Prevention for public health purposes—can alter children’s adrenocortical responses to acute stress. The behavioral and health consequences of this Pb-induced HPA dysregulation in children have yet to be determined.

Background
Several epidemiologic studies have suggested an association between Parkinson’s disease (PD) and exposure to heavy metals using subjective exposure measurements.

Objectives
We investigated the association between objective chronic occupational lead exposure and the risk of PD.

Methods
We enrolled 121 PD patients and 414 age-, sex-, and race-, frequency-matched controls in a case–control study. As an indicator of chronic Pb exposure, we measured concentrations of tibial and calcaneal bone Pb stores using 109Cadmium excited K-series X-ray fluorescence. As an indicator of recent exposure, we measured blood Pb concentration. We collected occupational data on participants from 18 years of age until the age at enrollment, and an industrial hygienist determined the duration and intensity of environmental Pb exposure. We employed physiologically based pharmacokinetic modeling to combine these data, and we estimated whole-body lifetime Pb exposures for each individual. Logistic regression analysis produced estimates of PD risk by quartile of lifetime Pb exposure.

Results
Risk of PD was elevated by > 2-fold [odds ratio = 2.27 (95% confidence interval, 1.13–4.55); p = 0.021] for individuals in the highest quartile for lifetime lead exposure relative to the lowest quartile, adjusting for age, sex, race, smoking history, and coffee and alcohol consumption. The associated risk of PD for the second and third quartiles were elevated but not statistically significant at the α = 0.05 level.

Conclusions
These results provide an objective measure of chronic Pb exposure and confirm our earlier findings that occupational exposure to Pb is a risk factor for PD.

Background

Outdoor air pollution and lead exposure can disturb cardiac autonomic function, but the effects of both these exposures together have not been studied.

Methods

We examined whether higher cumulative lead exposures, as measured by bone lead, modified cross-sectional associations between air pollution and heart rate variability among 384 elderly men from the Normative Aging Study. We used linear regression, controlling for clinical, demographic, and environmental covariates.

Results

We found graded, significant reductions in both high-frequency and low-frequency powers of heart rate variability in relation to ozone and sulfate across the quartiles of tibia lead. Interquartile range increases in ozone and sulfate were associated respectively, with 38% decrease (95% confidence interval = -54.6% to -14.9%) and 22% decrease (-40.4% to 1.6%) in high frequency, and 38% decrease (-51.9% to -20.4%) and 12% decrease (-28.6% to 9.3%) in low frequency, in the highest quartile of tibia lead after controlling for potential confounders. We observed similar but weaker effect modification by tibia lead adjusted for education and cumulative traffic (residuals of the regression of tibia lead on education and cumulative traffic). Patella lead modified only the ozone effect on heart rate variability.

Conclusions

People with long-term exposure to higher levels of lead may be more sensitive to cardiac autonomic dysfunction on high air pollution days. Efforts to understand how environmental exposures affect the health of an aging population should consider both current levels of pollution and history of lead exposure as susceptibility factors.

Lead is a confirmed neurotoxin, but questions remain about lead-associated intellectual deficits at blood lead levels < 10 μg/dL and whether lower exposures are, for a given change in exposure, associated with greater deficits. The objective of this study was to examine the association of intelligence test scores and blood lead concentration, especially for children who had maximal measured blood lead levels < 10 μg/dL. We examined data collected from 1,333 children who participated in seven international population-based longitudinal cohort studies, followed from birth or infancy until 5–10 years of age. The full-scale IQ score was the primary outcome measure. The geometric mean blood lead concentration of the children peaked at 17.8 μg/dL and declined to 9.4 μg/dL by 5–7 years of age; 244 (18%) children had a maximal blood lead concentration < 10 μg/dL, and 103 (8%) had a maximal blood lead concentration < 7.5 μg/dL. After adjustment for covariates, we found an inverse relationship between blood lead concentration and IQ score. Using a log-linear model, we found a 6.9 IQ point decrement [95% confidence interval (CI), 4.2–9.4] associated with an increase in concurrent blood lead levels from 2.4 to 30 μg/dL. The estimated IQ point decrements associated with an increase in blood lead from 2.4 to 10 μg/dL, 10 to 20 μg/dL, and 20 to 30 μg/dL were 3.9 (95% CI, 2.4–5.3), 1.9 (95% CI, 1.2–2.6), and 1.1 (95% CI, 0.7–1.5), respectively. For a given increase in blood lead, the lead-associated intellectual decrement for children with a maximal blood lead level < 7.5 μg/dL was significantly greater than that observed for those with a maximal blood lead level ≥7.5 μg/dL (p = 0.015). We conclude that environmental lead exposure in children who have maximal blood lead levels < 7.5 μg/dL is associated with intellectual deficits.

Abstract

Breast cancer (BC) is linked to estrogen exposure. Estradiol (E₂) stimulates BC cells proliferation by binding the estrogen receptor (ER). Hormone-related cancers have been linked to estrogenic environmental contaminants. Cadmium (Cd) a toxic pollutant, acts as estrogens in BC cells. Purpose of our study was to evaluate whether Cd regulates MCF-7 cell proliferation by activating ERK1/2, Akt and PDGFRα kinases. Cd increased cell proliferation and the ER-antagonist ICI 182,780 blunted it. To characterize an ER-dependent mechanism, ERα/β expression was evaluated. Cd decreased ERα expression, but not ERβ. Cd also increased ERK1/2, Akt and PDGFRα phosphorylation while ICI blocked it. Since stimulation of phosphorylation was slower than expected, c-fos and c-jun proto-oncogenes, and PDGFA were analyzed. Cd rapidly increased c-jun, c-fos and PDGFA expression. Cells were also co-incubated with the Cd and specific kinases inhibitors, which blocked the Cd-stimulated proliferation.

In conclusion, our results indicate that Cd increases BC cell proliferation in vitro by stimulating Akt, ERK1/2 and PDGFRα kinases activity likely by activating c-fos, c-jun and PDGFA by an ERα-dependent mechanism.

Abstract

Oxygen-free radicals, more generally known as reactive oxygen species (ROS) along with reactive nitrogen species (RNS) are well recognised for playing a dual role as both deleterious and beneficial species. The “two-faced” character of ROS is substantiated by growing body of evidence that ROS within cells act as secondary messengers in intracellular signalling cascades, which induce and maintain the oncogenic phenotype of cancer cells, however, ROS can also induce cellular senescence and apoptosis and can therefore function as anti-tumourigenic species. The cumulative production of ROS/RNS through either endogenous or exogenous insults is termed oxidative stress and is common for many types of cancer cell that are linked with altered redox regulation of cellular signalling pathways. Oxidative stress induces a cellular redox imbalance which has been found to be present in various cancer cells compared with normal cells; the redox imbalance thus may be related to oncogenic stimulation. DNA mutation is a critical step in carcinogenesis and elevated levels of oxidative DNA lesions (8-OH-G) have been noted in various tumours, strongly implicating such damage in the etiology of cancer. It appears that the DNA damage is predominantly linked with the initiation process. This review examines the evidence for involvement of the oxidative stress in the carcinogenesis process. Attention is focused on structural, chemical and biochemical aspects of free radicals, the endogenous and exogenous sources of their generation, the metal (iron, copper, chromium, cobalt, vanadium, cadmium, arsenic, nickel)-mediated formation of free radicals (e.g. Fenton chemistry), the DNA damage (both mitochondrial and nuclear), the damage to lipids and proteins by free radicals, the phenomenon of oxidative stress, cancer and the redox environment of a cell, the mechanisms of carcinogenesis and the role of signalling cascades by ROS; in particular, ROS activation of AP-1 (activator protein) and NF-κB (nuclear factor kappa B) signal transduction pathways, which in turn lead to the transcription of genes involved in cell growth regulatory pathways. The role of enzymatic (superoxide dismutase (Cu, Zn-SOD, Mn-SOD), catalase, glutathione peroxidase) and non-enzymatic antioxidants (Vitamin C, Vitamin E, carotenoids, thiol antioxidants (glutathione, thioredoxin and lipoic acid), flavonoids, selenium and others) in the process of carcinogenesis as well as the antioxidant interactions with various regulatory factors, including Ref-1, NF-κB, AP-1 are also reviewed.

OBJECTIVES: High levels of transition metals such as iron, nickel, chromium, copper, and lead are closely related to free radical generation, lipid peroxidation, formation of DNA strand breaks, and tumor growth in cellular systems. In order to determine the correlation to malignant growth in humans, we investigated the accumulation of heavy metals in 20 breast cancer biopsies and compared the findings to the levels found in 8 healthy biopsies. METHODS: The concentration of transition metals in breast cancer and control biopsies was assessed by a standardized Atomic Absorption Spectrofotometry technique with acidic hydrolysis for sample preparation. Additionally, heavy metal analysis in control biopsies was also performed with an Inductive Coupled Plasma--Mass Spectroscopy technique. For statistical analysis of the results, the Mann-Whitney U Test was applied. RESULTS: A highly significant accumulation of iron (p<0.0001), nickel (p<0.00005), chromium (p<0.00005), zinc (p<0.00001), cadmium (p<0.005), mercury (p<0.005), and lead (p< 0.05) was found in the cancer samples when compared to the control group. Copper and silver showed no significant differences to the control group, whereas tin, gold, and palladium were not detectable in any biopsies.
CONCLUSIONS: The data suggest that pathological accumulation of transition metals in breast tissue may be closely related to the malignant growth proess.

Soon after the exciting discovery of a method to transform human skin cells into stem cells in 2007 came the frustration of actually trying to make a sufficient amount of these induced pluripotent stem (iPS) cells.* The process is so inefficient that scientists typically only get 0.01 percent of a sample of human skin, or fibroblast, cells to form iPS cell colonies after they infect fibroblasts with the retroviruses used to induce pluripotency. "We almost gave up three years ago," says Dr. Duanqing Pei, director general and professor at the Guangzhou Institutes of Biomedicine and Health in Guangzhou, China.

But Pei's group at the Chinese Academy of Sciences plugged away and now has found that a simple chemical can give iPS cell transformation a big boost. By adding vitamin C to the medium in which human skin cells are grown, the researchers managed to turn 1 to 2 percent, compared to 0.01 percent, of cells into iPS cell colonies. "Once you reach the single digit frequency, that is no longer an infrequent event," Pei says. This yield, he believes, could allow scientists to finally study what is happening inside the cells as they undergo transformation. The results were published December 24 in Cell Stem Cell.

The ultimate goal is to make iPS cells a valid therapeutic option by replacing the potentially dangerous retroviruses with a safer cocktail of proteins and chemicals, including vitamin C.

The path that Pei's group took to identifying the beneficial role of vitamin C started with the realization that the factors that induce cells to become pluripotent were causing the cells to make the free radicals known as reactive oxygen species (ROS). "A high level of ROS is definitely very bad for the fibroblasts," Pei says, because it induces cell death at a faster rate.

To enhance the survival of cells and their likelihood of becoming iPS cells, Pei's team tested a variety of chemicals with anti-oxidant properties. The researchers started with fibroblast cells from mouse embryos, which have been shown to revert to iPS cells at a higher frequency than human fibroblasts. First, they infected the cells with retroviruses that trigger the expression of four embryonic cell-specific genes in the cells. Then they let the cells grow, either with or without vitamin C.

Ten days later, when cells typically begin to be reprogrammed into iPS cells, Pei's group found that 30 percent more mouse cells were alive in the Petri dish that contained vitamin C than in the one that did not, suggesting that the vitamin helps keep cells alive. One possible mechanism to explain this improved survival is that the cells exposed to ascorbic acid had lower levels of a tumor suppressor protein called p53, which can elicit cell death.

Surprisingly, the researchers found that vitamin C not only helps cells survive, but it enhances their progression to pluripotency by a factor of 100 . After 14 days, when cells start to become fully pluripotent, 10 to 20 percent of the mouse cells that were grown with vitamin C expressed genes associated with pluripotency compared with only 0.1 to 0.2 percent of the colonies grown without vitamin C. The group saw a similar improvement in human fibroblast reprogramming, from 0.01 percent in the absence of vitamin C to 1 percent in the presence. When other anti-oxidants were tested, none boosted the development of pluripotency.

Pei believes that ascorbic acid is spurring the induction of pluripotency in mouse and human cells through both its anti-oxidant properties as well as an as-yet unknown mechanism. Too much ROS and the reprogramming "machine" won't start, he says. But once ROS is reduced, the machine starts, and vitamin C makes it run more smoothly. "If you have too much ROS, the whole [reprogramming] machine will not move, but then once you [reduce] ROS, I think vitamin C does something [else] to make the machine move more smoothly," he says.

"Overall I think this is quite impressive progress," says Kwang-Soo Kim, director of the Molecular Neurobiology Lab at Harvard Medical School. Kim has developed a system to improve the safety of iPS cells by delivering the four proteins that induce pluripotency directly into cells. Because it avoids retroviruses, which integrate their genomes into cell chromosomes, his system carries less risk of potentially cancer causing mutations. But the efficiency is low—one to 10 percent of the viral method—and it's much slower. "We need a different kind of approach," he says.
This approach could be adding the four pluripotency-inducing proteins to cells to jumpstart the reprogramming "machine" and then improving the efficiency with a combination of chemicals, including vitamin C. Pei is currently trying to optimize the medium in which cells are grown. Although he doubts that any other anti-oxidant will have the same effect as ascorbic acid, Pei thinks that certain growth factors could further improve the culture conditions. In fact, his group found that combining vitamin C with valproic acid, which helps induce pluripotency, can improve transformation efficiency of human fibroblasts from 1 to 6 percent.

This level of efficiency could be enough to advance studies with iPS cells. "We don't need to generate 50 percent of the cells…as long as we can reproducibly generate a sufficient number of iPS lines," Kim says, adding that a 1 percent transformation efficiency could be enough.

Eventually, researchers would have to differentiate the iPS cells into certain cell types, says Kim, who himself has differentiated the more controversial embryonic stem cells into neurons to try to treat Parkinson's disease.

But, first, several studies with these vitamin C-induced pluripotent cells should be done, Kim notes. One possible problem – vitamin C causes cells to express lower levels of p53, which is important for the repair of DNA damage. Although Pei's group did not find any chromosomal abnormalities in cells grown with vitamin C, Kim says that higher resolution analysis is needed to ensure there are no mutations.

Because of the relatively high yield of Pei's method, these analyses and other studies of iPS cells should be possible. Now researchers might be able to generate enough cells to study the mechanism of and improve the safety and efficacy of iPS cells. "It's a worldwide effort to boost efficiency and make this more practical for much wider participation from the scientific community," Pei says.

* Note (12/29/09): This sentence was edited after publication to correct the year of the first human iPS cells.

Abstract

Cardiovascular disease is the leading cause of mortality worldwide. Arsenic is a ubiquitous metalloid in the crust of the earth. Chronic arsenic poisoning is becoming an emerging epidemic in Asia. Epidemiological studies have shown that chronic arsenic poisoning through ingestion of arsenic-contaminated water is associated with various cardiovascular diseases in dose–response relationships. These cardiovascular disorders include carotid atherosclerosis detected by ultrasonography, impaired microcirculation, prolonged QT interval and increased QT dispersion in electrocardiography, and clinical outcomes such as hypertension, blackfoot disease (a unique peripheral vascular disease endemic in southwestern Taiwan), coronary artery disease and cerebral infarction. Chronic arsenic poisoning is an independent risk factor for cardiovascular disease. The adverse cardiovascular effects of long-term arsenic exposure may be persistent and/or irreversible. Arsenic-induced cardiovascular diseases in human population may result from the interaction among genetic, environment and nutritional factors. The major adverse cardiovascular effect of chronic arsenic poisoning has been established qualitatively and quantitatively in the high arsenic exposure areas, but the low-dose effect of arsenic on cardiovascular diseases remains to be explored. Cardiovascular death is the major cause of mortality worldwide, and a small increased risk may imply a large quantity of excess mortality.

U.S. minority infants are born carrying hundreds of chemicals in their bodies, according to a report released today by an environmental group.

The Environmental Working Group's study commissioned five laboratories to examine the umbilical cord blood of 10 babies of African-American, Hispanic and Asian heritage and found more than 200 chemicals in each newborn.

"We know the developing fetus is one of the most vulnerable populations, if not the most vulnerable, to environmental exposure," said Anila Jacobs, EWG senior scientist. "Their organ systems aren't mature and their detox methods are not in place, so cord blood gives us a good picture of exposure during this most vulnerable time of life."

Of particular concern to Jacobs: 21 newly detected contaminants, including the controversial plastics additive bisphenol A, or BPA, which mimics estrogen and has been shown to cause developmental problems and precancerous growth in animals. Last month, researchers reported that male Chinese factory workers exposed to high levels of the chemical experienced erectile dysfunction and other sexual problems.

"BPA is a really important finding because people are really aware about its potential toxicity," Jacobs told reporters. "This is the first study to find BPA in umbilical cord blood, and it correlates with national data on it."

Jacobs said the study focused on minority children to show that chemical exposure is ubiquitous, building on 2005 research on cord blood from 10 anonymous babies. That study found a similar body burden among the babies. This is the first study to look at chemicals in minority newborns.

"Minority groups may have increased exposure to certain chemicals, but here we didn't focus on those chemicals," Jacobs said. "The sample size is too small to see major differences, but we want to increase awareness about chemical exposures."

Leo Trasande, co-director of the Children's Environmental Health Center at the Mount Sinai School of Medicine, said the findings, while preliminary, show that minority communities are often disproportionately affected by chemical exposure. Trasande was not involved in the EWG study.

"Presently, minority communities suffer from a host of chronic disorders, and disproportionate chemical exposures may contribute significantly to the origins of the disparities that exist," Trasande said.

Both he and Jacobs said the findings add momentum for the call to revamp the Toxic Substances Control Act, or TSCA, the law regulating the more than 80,000 chemicals on its database. They released the report on the same day that a Senate panel is scheduled to discuss the government's strategy for managing the tens of thousands of chemicals in the marketplace with an eye toward overhauling TSCA.

TSCA does not require most chemicals to be tested for safety before they are approved for widespread use. Because of this, Trasande said, less than half of the 3,000 high-production volume chemicals on the marketplace have toxicity data, and less than one-fifth have toxicity testing data on the effects on developing organs.

"These results are alarming for their implications of health impacts on children," Trasande said.

Another challenge facing chemical regulators is understanding how the different chemicals interact together, which is particularly significant given the number of chemicals found in people.

Objectives. We investigated whether foreign birthplace and residence were associated with an increased risk of childhood lead poisoning.

Methods. We conducted a matched case–control study among New York City children (mean age=3 years) tested for lead poisoning in 2002 (n=203 pairs). Children were matched on age, date of test, and residential area. Blood lead and housing data were supplemented by a telephone survey administered to parents or guardians. Conditional logistic regression analysis was used to examine the relationship of lead poisoning status to foreign birthplace and time elapsed since most recent foreign residence after adjustment for housing and behavioral risk factors.

Results. Both foreign birthplace and time since most recent foreign residence had strong adjusted associations with lead poisoning status, with children who had lived in a foreign country less than 6 months before their blood test showing a particularly elevated risk of lead poisoning relative to US-born children with no foreign residential history before their blood test (odds ratio [OR]=10.9; 95% confidence interval [CI]=3.3, 36.5).

Conclusions. Our findings demonstrate an increased risk of lead poisoning among immigrant children.

Persons employed in high-risk lead-related occupations can transport lead dust home from a worksite through clothing, shoes, tools, or vehicles (1-4). During 2008, the Maine Childhood Lead Poisoning Prevention Program (MCLPPP) identified 55 new cases of elevated (≥15 µg/dL) venous blood lead levels (BLLs) among children aged <6 years through mandated routine screening (5,6). Although 90% of childhood lead poisoning cases in Maine during 2003--2007 had been linked to lead hazards in the child's home, no lead-based paint or dust or water with elevated lead levels were found inside the homes associated with six of the 2008 cases (i.e., five families, including one family with two affected siblings). An expanded environmental investigation determined that these six children were exposed to lead dust in the family vehicles and in child safety seats. The sources of the lead dust were likely household contacts who worked in high-risk lead exposure occupations. Current recommendations for identifying and reducing risk from take-home lead poisoning include 1) ensuring that children with elevated BLLs are identified through targeted blood lead testing, 2) directing prevention activities to at-risk workers and employers, and 3) improving employer safety protocols. State and federal prevention programs also should consider, when appropriate, expanded environmental lead dust testing to include vehicles and child safety seats.

Lead poisoning has decreased among children in the United States because of federal, state, and community efforts to reduce exposure (7). Federal bans on leaded gasoline and lead-based paint, and improvements in occupational safety and health standards* have helped mitigate exposure to lead, especially among children. MCLPPP responds to all reported elevated blood lead levels ≥10 µg/dL. Children with venous BLLs ≥15µg/dL automatically trigger an environmental investigation to determine the lead sources, and children are monitored until their venous BLLs are <10µg/dL.

For this study, a case of lead poisoning was defined by a confirmed venous BLL ≥15 µg/dL in a child aged <6 years living in Maine. All cases were identified through mandated blood lead testing for children at ages 1 year and 2 years following CDC targeted lead testing recommendations (5,6).† A case of take-home lead poisoning was defined by 1) a confirmed venous BLL ≥15 µg/dL among children aged <6 years living in Maine, 2) a household contact in a high-risk lead-related occupation, and 3) environmental lead dust sampling of vehicle and child safety seat ≥40 µg/ft2, with no detectable lead-based paint hazards present in the home.

When these investigations began, MCLPPP contacted each child's family and offered general lead education, nursing case management, and environmental lead investigations by licensed lead risk assessors to determine the likely sources of the poisoning. Families were interviewed using a MCLPPP risk-assessment questionnaire to determine other possible exposures. Radiograph fluorescence analysis was used to determine whether lead-based paint was in the homes. Lead dust wipe samples were taken using the Environmental Protection Agency (EPA) standard lead dust loading methodology in the homes.§ For the cases described in this report, MCLPPP also directed investigators to perform additional dust sampling in the family vehicles and child safety seats because household members had occupations at high risk for lead exposure. The EPA acceptable lead dust standard is <40 µg/ft2 for floors inside the home, but no lead standards have been set for vehicles or child safety seats.

The six children with take-home lead poisoning, including two siblings in one family, ranged in age from 4 to 28 months, and had a median venous BLL of 21 µg/dL (range: 15--32 µg/dL). Among the five families, contacts included four persons who currently or recently worked in painting and paint removal, and one who was a self-employed metals recycler. The workers reported no lead-related occupational safety measures provided by their employers at work sites.

Four of the five homes were built after 1978, the year lead-based paint was banned. No lead-based paint was detected by radiograph fluorescence analysis inside the five homes. In two of five homes, lead dust was detected in exterior areas where family members removed and kept work clothes, including an entryway/deck (110 µg/ft2), another entryway (1,200 µg/ft2), and a laundry room (40 µg/ft2). Five family vehicles (one family did not own a vehicle and one family had two) tested positive for lead dust with a median of 550 µg/ft2 for driver/passenger seats (range: 49--2,100 µg/ft2) and a median of 1,570 µg/ft2 for driver/passenger floors (range: 240--2,900 µg/ft2). All child safety seats (n = 6) tested positive for lead dust with a median of 98 µg/ft2 (range: 43--420 µg/ft2). Three safety seats were stored in the vehicle (median lead dust: 120 µg/ft2 [range: 43--420 µg/ft2]); the other three were removed and kept in the home when not in use (median lead dust: 95 µg/ft2 [range: 50--100 µg/ft2]).

MCLPPP determined that the primary source of lead exposure was lead dust in the family vehicles and on the child safety seats (Table), and provided recommendations to prevent continued exposure. Persons who are exposed to lead at work or through hobbies are advised upon finishing the workday to 1) place lead-contaminated clothes, including shoes and personal protective equipment, in a closed container for laundering or cleaning; 2) take a shower and wash hands, face, and hair when exposed above the permissible exposure limits; 3) change into street clothes; and 4) wash work clothes separately from all other clothes.** However, parents and household contacts reported a lack of facilities available for washing, showering, and changing clothes before entering their personal vehicles. MCLPPP also recommended thorough vacuuming and wet cleaning of the vehicle interiors and replacement of any child safety seat that tested positive for lead dust. Families were referred to the Maine Injury Prevention Program for replacement safety seats, if needed.

Abstract

Perinatal exposure to cadmium (Cd) or methylmercury (MeHg) results in impaired neurodevelopment. Thyroid hormone is essential for normal brain development. However, the issue whether Cd or MeHg, especially at low doses, interrupts thyroid hormone action remains to be investigated. In the present study, effects of perinatal exposure to low levels of Cd or MeHg on thyroid hormone metabolism were examined using metallothionein I and II (MT-I/II) null or wild-type neonatal mice. Dams were exposed to 10 mg/L water of Cd or 5 mg/kg chow of MeHg from gestational day 0 to post-natal day 10 (PND 10). Sera, livers and brains were collected from neonates on PND 10. Iodothyronine deiodinase activities and serum thyroxine (T4) concentrations were measured. MeHg exposure failed to induce changes in serum T4 levels and liver type 1 deiodinase (D1) and brain type 2 deiodinase (D2) activities regardless of the MT genotype. However, exposure to MeHg resulted in a decrease in brain type 3 deiodinase (D3) activity in MT-I/II null and wild-type neonates. In contrast, exposure to Cd resulted in a decrease in serum T4 levels in MT-I/II null neonates. Consistently, brain D2 activity was increased in Cd-exposed MT-I/II null neonates. No significant changes in liver D1 and brain D3 activities were induced by Cd administration. Our study demonstrates that perinatal exposure to low doses of Cd or MeHg can induce changes in brain deiodinase activities in the neonates, suggesting that thyroid hormone metabolism in fetuses and neonates might be a potential target of Cd and MeHg.

Abstract
Chemical exposure can trigger or accelerate the development of autoimmune manifestations. Although heavy metals are elementary chemical structures, they can have profound and complex effects on the immune system. In genetically susceptible mice or rats, administration of subtoxic doses of mercury induces both the production of highly specific autoantibodies and a polyclonal activation of the immune system. We review in this article some of the mechanisms by which heavy metal exposure can lead to autoimmunity.

Abstract
This study evaluated the relationship between pregnancy rate and semen cadmium concentration. This prospective and nonrandomized clinical study analyzed 341 male partners of infertile couples undergoing infertility evaluation and management. Semen samples were collected to analyze semen quality and cadmium concentrations. The main outcome was pregnancy during 60-day infertility treatment. Simple linear regression analysis revealed an association between semen cadmium concentration NS sperm count (r = −0.150, P = 0.0416) in nonsmoking subjects (n = 184). In both smokers and nonsmokers, semen cadmium concentrations were significantly higher in non-pregnant patients than in pregnant patients. In nonsmokers, Cox multi-variable fertility ratio analysis demonstrated an association between semen cadmium concentration and fertility (fertility ratio of log semen cadmium = 0.24; 95% confidence intervals (CI) = 0.12–0.47, P < 0.0001) after adjusting for related variables. Each tenfold increase in semen cadmium concentration was associated with a 4.17-fold increase in infertility ratio in nonsmoking patients. In smokers, Cox multi-variable fertility ratio analysis demonstrated that sperm count and semen cadmium concentration are associated with fertility (fertility ratio of log semen cadmium = 0.17; 95% CI = 0.04–0.63, P = 0.0085) after adjusting for related variables. In smokers, each tenfold increase in semen cadmium concentration was associated with a 5.88-fold increase in infertility ratio. In conclusion, low levels of cadmium accumulation in semen may contribute to male infertility by reducing sperm quality.

BACKGROUND: It has been hypothesized that exposure to exogenous estrogens may be associated with endometriosis and uterine myomas. We sought to investigate the association between heavy metals which have been shown to be hormonally active and these disorders using data from the National Health and Nutrition Examination Survey, 1999–2002.

METHODS: Women aged 20–49 years who had data on metals and the outcomes of interest, were premenopausal and neither pregnant nor breastfeeding were eligible (n = 1425). Lead, cadmium and mercury were measured in whole blood. Diagnosis of outcomes was based upon self-report. Logistic regression was used to examine the association between tertiles of heavy metals and disease adjusting for age, race/ethnicity, use of birth control pills prior to diagnosis and smoking status at diagnosis.

RESULTS: A dose–response association between cadmium and endometriosis was observed [tertile 2 versus 1: adjusted odds ratio (OR) = 1.94, 95% confidence interval (CI): 0.73–5.18; tertile 3 versus 1: adjusted OR = 3.39, 95% CI 1.37–8.40]. This association persisted in subanalyses: (i) limiting analysis to women diagnosed in the past 10 years and (ii) limiting analysis to women diagnosed since last pregnancy, although limited by sample size.

CONCLUSIONS: These results must be interpreted with caution given the cross-sectional study design. The observed association between cadmium and endometriosis deserves further investigation in properly designed studies.

Objective
This systematic review evaluates the evidence on the association between lead exposure and cardiovascular end points in human populations.

Methods
We reviewed all observational studies from database searches and citations regarding lead and cardiovascular end points.

Results
A positive association of lead exposure with blood pressure has been identified in numerous studies in different settings, including prospective studies and in relatively homogeneous socioeconomic status groups. Several studies have identified a dose–response relationship. Although the magnitude of this association is modest, it may be underestimated by measurement error. The hypertensive effects of lead have been confirmed in experimental models. Beyond hypertension, studies in general populations have identified a positive association of lead exposure with clinical cardiovascular outcomes (cardiovascular, coronary heart disease, and stroke mortality; and peripheral arterial disease), but the number of studies is small. In some studies these associations were observed at blood lead levels < 5 μg/dL.

Conclusions
We conclude that the evidence is sufficient to infer a causal relationship of lead exposure with hypertension. We conclude that the evidence is suggestive but not sufficient to infer a causal relationship of lead exposure with clinical cardiovascular outcomes. There is also suggestive but insufficient evidence to infer a causal relationship of lead exposure with heart rate variability.

Public Health Implications
These findings have immediate public health implications. Current occupational safety standards for blood lead must be lowered and a criterion for screening elevated lead exposure needs to be established in adults. Risk assessment and economic analyses of lead exposure impact must include the cardiovascular effects of lead. Finally, regulatory and public health interventions must be developed and implemented to further prevent and reduce lead exposure.

Abstract

Epstein-Barr virus (EBV) hepatitis is an uncommon, almost always self-limited disease in immunocompetent patients. Accurate diagnosis is imperative for appropriate clinical management. The aim of this study was to compare 3 available methods for EBV detection on routinely processed liver biopsies to determine their effectiveness in aiding the diagnosis. In 6 of the 8 cases of EBV hepatitis, EBV was detected by both polymerase chain reaction (PCR) for EBV DNA and in situ hybridization (ISH) for EBV early RNA (EBER). EBV was detected by PCR only in 1 case, and by ISH only in another. EBER-positive cells detected by ISH were typically few and individually distributed in the portal tracts and sinusoids. Immunohistochemical staining for EBV latent membrane proteins was negative in all 8 cases. Five cases of chronic hepatitis C used as negative controls were negative by all 3 detection methods for EBV. These data indicate that PCR and ISH are equally sensitive in detecting EBV in routinely processed liver biopsies. The ready implementation of ISH in pathology laboratories makes it a useful ancillary tool in confirming the diagnosis of EBV hepatitis in equivocal cases. However, EBER-positive cells can be sparse and easily overlooked. Immunohistochemistry for EBV latent membrane proteins apparently has no utility in the diagnosis of EBV hepatitis.

Abstract
As the pathogenesis of fibromyalgia continues to raise debate, multiple putative triggers have been implicated. The current review summarizes the available data linking fibromyalgia to either infection or vaccination. Multiple infectious agents have been associated with the development of either full-blown fibromyalgia (e.g. hepatits C), or with symptom complexes extensively overlapping with that syndrome (e.g. chronic Lyme disease). The cases of Lyme disease, mycoplasma, hepatits C and HIV are detailed. Despite the described associations, no evidence is available demonstrating the utility of antibiotic or anti-viral treatment in the management of fibromyalgia. Possible mechanistic links between fibromyalgia and HIV are reviewed. Associations have been described between various vaccinations and symptom complexes including fibromyalgia and chronic fatigue syndrome. The case of Gulf War syndrome, a functional multisystem entity sharing many clinical characteristics with fibromyalgia is discussed, with emphasis on the possibility of association with administration of multiple vaccinations during deployment in the Persian Gulf and the interaction with stress and trauma. Based on this example a model is proposed, wherein vaccinations function as co-triggers for the development of functional disorders including fibromyalgia, in conjunction with additional contributing factors.

Objective

To describe a case of bilateral acute retinal necrosis syndrome (ARNS) in a 5-year-old boy.

Method

A retrospective, interventional case is described in one child attending the pediatric ophthalmology section, complaining of sudden bilateral red eye and haze-impaired vision. A standardized ophthalmologic examination and specific serological probes supported the diagnosis of severe bilateral ARNS in an immunocompetent child.

Results

The reduced visual acuity (<20/400), the ocular fundus signs (perivasculitis, thrombosis and retinal edema) and the positive immunoglobulin M anti-Epstein Barr virus serology, lead us to the ARNS definitive diagnosis. Antiviral therapy (Acyclovir; Zovirax®), ciclopentolate dilating eye drops, and antiplatelet treatment (acetil salicylic acid; Aspirin®) were administered until recovering the final visual acuity (20/40).

Conclusions

The ARNS is an ocular disease with poor prognosis, which in turns may display better course when determining the etiopathogenic virus and selecting the appropriate and precocious therapy.

The objective of this study was to assess the incidence and the clinical significance of Epstein-Barr virus (EBV) in patients with chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL). Patients with CLL/SLL who presented at The University of Texas M. D. Anderson Cancer Center over a 2-year period and had available marrow paraffin blocks were studied for evidence of EBV infection using a highly specific in-situ hybridization assay for detection of EBV encoded RNA (EBERs). Results were analysed in relation to other presenting characteristics and outcome. Thirty-two patients were examined. EBERs were detected in the bone marrow of 12 of 32 (38%) CLL/SLL marrows vs 0 of 20 normal marrows (p = 0.002). EBERs were observed in sporadic granulocytes alone or in addition to its presence in lymphocytes in nine of the 12 EBV-positive patients. EBERs were detected less frequently in patients with Rai stage 0 – 1 disease (20%) compared with Rai stage 2 – 4 (66%; p = 0.008). EBER-positive patients tended to have higher lactate dehydrogenase levels (p = 0.053). The 10-year survival rate was 22% vs 58% for patients with and without discernible EBERs (log-rank, p = 0.08). Evidence of EBV infection was found in 38% of patients with CLL/SLL. Despite the small number of patients tested, discernable EBERs were significantly more common in individuals with more advanced Rai stage and there was a trend toward shorter survival in patients in whom EBV EBERs were discerned. Larger studies are needed to determine the prognostic value and role of EBV infection in patients with CLL/SLL.

Introduction
Epstein-Barr virus (EBV) is present in over 90% of the world's population. This infection is considered benign, even though in limited cases EBV is associated with infectious and neoplastic conditions. Over the past decade, the EBV association with breast cancer has been constantly debated. Adding to this clinical and biological uncertainty, different techniques gave contradictory results for the presence of EBV in breast carcinoma specimens. In this study, minor groove binding (MGB)-TaqMan real time PCR was used to detect the presence of EBV DNA in both peripheral blood and tumor samples of selected patients.

Methods
Peripheral blood and breast carcinoma specimens from 24 patients were collected. DNA was extracted and then amplified by MGB-TaqMan real time PCR.

Results
Of 24 breast tumor specimens, 11 (46%) were positive for EBV DNA. Of these 11 breast tumor specimens, 7 (64%) were also positive for EBV DNA in the peripheral blood, while 4 (36%) were positive for EBV DNA in the tumor, but negative in the blood.

Conclusion
EBV was found at extremely low levels, with a mean of 0.00004 EBV genomes per cell (range 0.00014 to 0.00001 EBV genomes per cell). Furthermore, our finding of the presence of EBV in the tumor specimens coupled to the absence of detection of EBV genomic DNA in the peripheral blood is consistent with the epithelial nature of the virus. Because of the low levels of viral DNA in tumor tissue, further studies are needed to assess the biological input of EBV in breast cancer.

Most people are quite aware of the fact that vitamin supplements are a great idea to support the overall health and wellbeing of children and young adolescents. However, one of the vitamins that is sometimes overlooked is Vitamin D, a vitamin that is essential for bone health and may offer additional protection against childhood coughs and colds.

Children should be able to get all the Vitamin D they need from exposure to sunlight, but the lifestyle of many of today’s children and teens means that they simply do not spend enough time outside to gain the maximum benefits.

A new study recently published has revealed that levels of Vitamin D found amongst US children have indeed now fallen below what is considered medically acceptable, and that the situation amongst African American and Hispanic children is particularly dire.

The latest research was conducted by Jonathan Mansbach, MD and his team who are based at Children’s Hospital Boston. They used data culled from the National Health and Nutrition Examination Survey which encompasses medical data about approximately 5,000 children from across the United States and was collected between 2001-2006.

Taking the current recommendation from the American Academy of Pediatrics that children and teens maintain a 50 nmol/L level of Vitamin D in their system to be considered healthy the researchers found that approximately 20% of US children will fall below this guideline score and in the case of Hispanic children that figure rises to an astonishing 82%. The outlook is even worse for non Hispanic black children as the research suggests that 92% suffer from Vitamin D deficiency.

There are those who disagree with the APA guidelines, saying that other research in adults has suggested that 75 nmol/L is a more reasonable level to ensure good health. Should this eventually turn out to be the case then the picture is even grimmer. Says lead author of the study Mansbach “If 75 nmol/L or higher is eventually demonstrated to be the healthy normal level of vitamin D, then there is much more vitamin D deficiency in the U.S. than people realize”

The researchers conclude their report by recommending that all children and teens take a Vitamin D supplement on a daily basis to ensure that their bodies have enough Vitamin D to maintain healthy growth. It is especially crucial they note for those who live in high altitudes where sunlight is scarce in winter.

Background. In Western countries, the cause of 60% of all Guillain‐Barre syndrome (GBS) cases remains unidentified. The number of cases of unidentified cause peaks in winter, and these cases are commonly preceded by respiratory tract infection or influenza‐like illness. We investigated the triggering role of influenza virus infection.

Methods. Of 405 patients with GBS who were admitted to a French reference center during 1996–2004, 234 had cases caused by an unidentified agent. We used time‐series methods to study the correlation between the monthly incidence of such cases and influenza‐like illnesses reported by the Sentinelles surveillance network. We analyzed anti‐influenza antibodies using complement fixation testing and hemagglutination‐inhibition assays. We studied etiological subgroups using Wilcoxon and Fisher’s exact tests.

Results. We found a positive association between the monthly incidence of GBS caused by an unidentified agent and reported influenza‐like illnesses. Of 73 patients whose cases occurred during periods in which there was a possible link to influenza, 10 (13.7%) had serological evidence of recent influenza A, and 4 (5.5%) had serological evidence of influenza B. Eight of 10 influenza A–related cases occurred during “major” influenza seasons, and antibodies specific to the current epidemic strain were found in 9 cases. Most patients with influenza A–related cases were aged <65 years, and none had antiganglioside antibodies. Influenza‐related cases differed both from Campylobacter jejuni–related cases, with regard to the lack of need for mechanical ventilation ( P = .0,14), and from the cases caused by an unidentified agent, with regard to the presence of preceding influenza‐like illness or respiratory tract infection ( P = .0,15) and longer time from the infectious event to GBS onset (P = .0,4 ).

Conclusions. Influenza viruses are infrequent triggering agents of GBS but may play a significant role during major influenza outbreaks. Influenza‐related GBS displays specific features and is not associated with antiganglioside antibody response, which suggests the presence of underlying immune mechanisms.

Context  An unexplained increase in the risk of Guillain-Barre syndrome (GBS) occurred among recipients of the swine influenza vaccine in 1976-1977. Guillain-Barre syndrome remains the most frequent neurological condition reported after influenza vaccination to the Vaccine Adverse Events Reporting System (VAERS) since its inception in 1990.

Objective  To evaluate trends of reports to VAERS of GBS following influenza vaccination in adults.

Design, Setting, and Participants  VAERS is the US national spontaneous reporting system for adverse events following vaccination. Reports of GBS in persons 18 years or older following influenza vaccination were evaluated for each influenza season from July 1, 1990, through June 30, 2003. The number of people vaccinated was estimated from the National Health Interview Survey and US census data. Beginning in 1994, active follow-up was conducted to verify GBS diagnosis and obtain other clinical details.

Main Outcome Measure  Reporting rates of GBS following influenza vaccination over time.

Results  From July 1990 through June 2003, VAERS received 501 reports of GBS following influenza vaccination in adults. The median onset interval (13 days) was longer than that of non-GBS reports of adverse events after influenza vaccine (1 day) (P<.001). The annual reporting rate decreased 4-fold from a high of 0.17 per 100  000 vaccinees in 1993-1994 to 0.04 in 2002-2003 (P<.001). A GBS diagnosis was confirmed in 82% of reports. Preceding illness within 4 weeks of vaccination was identified in 24% of reported cases.

Conclusions  From 1990 to 2003, VAERS reporting rates of GBS after influenza vaccination decreased. The long onset interval and low prevalence of other preexisting illnesses are consistent with a possible causal association between GBS and influenza vaccine. These findi