Developed Skin Lesions Research Articles

Human DcR3 overexpression in mice results in a systemic lupus erythematosus-like syndrome (129.15)

Abstract OBJECTIVE: DcR3, a TNFR family member, is a secreted protein, and can enhance cell survival by interfering with multiply apoptosis pathways. The objective of this study was to understand the role of DcR3 in pathogenesis of autoimmune diseases. METHODS: We generated transgenic (Tg) mice with actin promoter-driven expression of human DcR3, and investigated the development of autoimmune diseases in these mice. RESULTS: Beyond 5–6 months of age, DcR3 Tg mice developed a systemic lupus erythematosus (SLE)-like syndrome. They produced autoantibodies against various organs, and against dsDNA and Smith Ag. The kidneys of these Tg mice showed pathological changes indicative of glomerular nephritis and IgG and C3 deposition; kidney dysfunctions, such as proteuria, leukocyturia, and hemuresis, were obvious. Aged Tg mice also developed skin lesions and lymphocyte infiltration in the liver, and suffered from leukopenia, anemia and thrombocytopenia. The SLE-like symdrome penetrance in DcR3 Tg mice was gender-dependent, with about 60% in females versus 20% in males. Mechanistically, exogenous recombinant DcR3 or endogenous DcR3 produced by Tg T cells effectively protected CD4 cells from activation-induced apoptosis in vitro. Probably as a consequence, in the peripheral blood of Tg mice beyond 6 months of age, CD4 cells with a phenotype of previous activation were increased. CONCLUSION: DcR3 overexpression could lead to a SLE-like syndrome in mice.

Susceptibility to arsenic-induced skin lesions from polymorphisms in base excision repair genes

Genetic polymorphisms in the base excision DNA repair pathway may influence individual susceptibility to arsenic and the development of arsenic-induced skin lesions. Data from a case-control study of 792 cases and 792 matched controls conducted in Bangladesh from 2001 to 2003 were analyzed using conditional logistic regression to assess the associations between four common base excision repair (BER) genetic polymorphisms X-ray repair cross-complementing group 1 (XRCC1) Arg399Gln, XRCC1 Arg194Trp, human 8-oxoguanine DNA glycosylase (hOGG1) Ser326Cys and apurinic/apyrimidinic endonuclease (APE1) Asp148Glu and arsenic-induced skin lesions including melanosis and keratosis. Adjusted for toenail arsenic, body mass index, education, smoking and betel nut use, individuals with the APE1 148Glu/Glu polymorphism had a 2-fold increased odds of skin lesions compared with individuals with the 148Asp/Asp genotype (1.93; 95% confidence interval 1.15, 3.19). Gene-environment interactions between toenail arsenic and XRCC1 Arg194Trp and APE1 Asp148Glu were observed. Within the lowest arsenic tertile, APE1 148Glu/Glu had 2.5 times the odds ratio compared with wild-type, whereas within the highest tertile of arsenic the odds ratios for skin lesions did not differ. In contrast, at low arsenic levels, the odds ratios for skin lesions did not differ much by XRCC1 Arg194Trp genotype. However, at the highest tertile of arsenic, the XRCC1 194Arg/Arg polymorphism conferred a 3-fold larger odds ratio for skin lesions compared with XRCC1 194Trp/Trp. Individuals may have different odds for developing skin lesions based in part on their genetic profile for BER and their arsenic exposure history. Future research on arsenic-induced skin lesions should consider the impact of genetic variation to individual susceptibility to arsenic toxicity.

Mycological evaluation of bronchoalveolar lavage in cats with respiratory signs from Rio de Janeiro, Brazil

Twenty-three cats with respiratory signs who had domiciliary contact with cats with sporotrichosis were studied. Sneezing was the predominant extracutaneous sign. Twelve cats had no skin lesions and 11 had ulcerated skin lesions. Mycological culture of material obtained from the nasal cavity, oral cavity, bronchoalveolar lavage (BAL) and skin lesions, when present, was performed for all cats. In the case of autopsy, lung fragments were cultured. Sporothrix schenckii was isolated from four of the 12 cats without skin lesions: BAL (one cat) and oral and/or nasal cavity (three cats). The latter three animals developed nasal and distant skin lesions within the following 2-4 weeks. The cat with S. schenckii isolated from BAL did not develop skin lesions or lower respiratory tract symptoms during the 6 months of follow-up. S. schenckii was isolated from one or more biological samples of all 11 cats with skin lesions: oral cavity (five), nasal cavity (eight), BAL fluid (four), skin lesions (eight), and blood culture (one). No yeast-like structures were observed upon BAL cytology in any of the 23 cats. The results suggest that S. schenckii can cause infection of skin contiguous to the natural facial orifices through colonisation of the mucosal surfaces of the upper airways.

Effect of substrate on progression and healing of skin erosions and epidermal papillomas of Atlantic halibut, Hippoglossus hippoglossus (L.)

Juvenile Atlantic halibut cultured on a smooth substrate often develop skin lesions on the blind (abocular) side, significantly reducing carcass value and causing economic losses. We discovered that fish not only developed skin erosions, but also extensive epidermal papillomas when held on a smooth substrate for 39 months. This is the first time that epidermal papillomas have been reported in Atlantic halibut. To determine whether substrate type affected the progression or healing of these lesions, fish with papillomas and skin erosions were moved to identical rearing units having either an irregular (sand, gravel or Netlon) substrate, or a smooth (gel-coated fibreglass) substrate. After 62 days, 42-50% of fish held on any of the three irregular substrates had a skin erosion area (SEA) that had either diminished or appeared to be completely healed via gross examination. In contrast, none of the fish held on the smooth substrate showed improvement and 75-100% had a more severe SEA. Changes in the papilloma area (PA) were less dramatic: none of the fish held on an irregular substrate had any clear change in PA, although there was a suggestion that the PA in some fish might becoming less severe. Similarly, none of the fish held on the smooth substrate had a change in PA, although there was a suggestion that the PA in some fish was becoming more severe. After 97 days, there was also a suggestion that specific growth rates were higher in fish held on an irregular substrate when compared with those held on the smooth substrate. Overall, these data further substantiate previous studies suggesting that an irregular substrate, including the commercially used Netlon, is best for rearing Atlantic halibut. While there was a clear relationship between substrate type and whether skin erosions or papillomas progressed or healed, the exact cause(s) of these two responses is uncertain. Future studies should focus on risk factors such as stress, burrowing behaviour and the microenvironment of the substrate.

HAMP AND HVJ AS CANDIDATE MODIFIER GENES IN TYPE 1 HEREDITARY HEMOCHROMATOSIS WITH HIGH IRON BURDEN AND IN PORPHYRIA CUTANEA TARDA.

Homozygosity for the HFE C282Y (HFE Y/Y) mutation accounts for approximately 90% of HFE-associated (type 1) hereditary hemochromatosis (hh), but the clinical phenotype ranges from simply an elevated transferrin (Tf) saturation to organ damage due to iron overload. Modifier genes have been proposed to explain this phenotypic variability. A second disease associated with hepatic iron overload is porphyria cutanea tarda (PCT). Approximately 20% of patients with PCT are HFE Y/Y, but the cause of hepatic iron loading in the remaining 80% is not known. Two genes known to affect iron homeostasis are hepcidin (HAMP) and hemojuvelin (HJV). Heterozygosity for HAMP and HJV mutations has been associated with iron overload in a small number of type 1 hh patients (Blood. 2004;103:2835-40; Blood Cells Mol Dis 2004;33:338-43). We asked if mutations of HAMP or HJV could account for hepatic iron overload in highly penetrant hh patients and in PCT patients with or without HFE mutations. Serum iron values did not differ between hh and PCT patients, but Tf saturation and serum ferritin values were higher in hh patients compared with PCT patients with hh. This may reflect the fact that PCT patients develop skin lesions leading to diagnosis at a younger age than organ damage occurs in patients with hh but without PCT. We sequenced the HAMP and HJV genes in 96 hh patients with grade 3 to 4 (scale 0-4) hepatic parenchymal cell stainable iron and 96 PCT patients with variable degrees of hepatic siderosis. Ninety-four percent (90) of the hh patients were HFE Y/Y, 4.2% (4) were HFE C/Y, and 2.1% (2) were wild-type HFE C/C. No exonic changes or splice site mutations were detected in either the HAMP or HJV genes. Eighty-three of the 96 PCT patients were genotyped. Twenty-five percent (21) were HFE Y/Y, 23% (19) were HFE C/Y, and 52% (43) were HFE C/C. No exonic changes or splice site mutations were detected in the HJV gene of patients with PCT, but two PCT patients were found to be heterozygotes for HAMP mutations. The first had the previously identified 212G→A transition leading to a G71D substitution. The second had a 248A→C transversion corresponding to K83R in the peptide. Both of these PCT patients were HFE C/C, but both had grade 4 HPCSI. These data indicate that heterozygosity for mutations of HAMP or HJV rarely modifies the iron loading phenotype in either type 1 hh or PCT. Other modifier loci must exist.

Rapidly growing collagenomas in multiple endocrine neoplasia type I

Patients with multiple endocrine neoplasia type I (MEN-I) frequently develop skin lesions including collagenomas, angiofibromas, and lipomas. We report a patient with MEN-I who exhibited rapid growth of multiple collagenomas after pancreatic enucleation of a vasoactive intestinal peptide-secreting tumor (VIPoma) and excision of multiple pancreatic masses. Five of the collagenomas were protuberant, with the bulk of the lesion protruding above the skin. Histologic analysis of the collagenomas revealed broad collagen bundles in a haphazard arrangement and decreased elastic fibers. Rapid growth of protuberant collagenomas appears to be unusual in MEN-I, but we suggest that MEN-I be considered in patients with apparent eruptive collagenoma.

Suppression of Skin Lesions by Transdermal Application of CpG-Oligodeoxynucleotides in NC/Nga Mice, a Model of Human Atopic Dermatitis

Atopic dermatitis (AD) is a pruritic inflammatory skin disease characterized by an elevation of the total IgE level in plasma, the infiltration of mast cells and eosinophils, and the expression of cytokines by Th2 cells. NC/Nga mice kept in conventional conditions are known to develop skin lesions resembling human AD. We examined in this study the alterations of immune response in NC/Nga mice kept in conventional conditions, following transdermal application of CpG-oligodeoxynucleotides (ODN), which plays a critical role in immunity via the augmentation of Th1-type and suppression of Th2-type responses. CpG-ODN remarkably changed the immune response from type Th2 to Th1 as determined from cytokine mRNA and Ab levels. The serum IgE level was decreased and the expression of IgG2a was up-regulated. The application of CpG-ODN to the skin also decreased inflammatory infiltration of mast cells, and suppression in the skin lesions was observed. Furthermore, the generation of regulatory T cells, which are considered immune suppressive T cells, was observed in the skin on treatment with CpG-ODN. These results suggested CpG-ODN is effective for immunotherapy in patients with AD, which is characterized by Th2-dominated inflammation.

Runx3 regulates dendritic epidermal T cell development

The Runx3 transcription factor regulates development of T cells during thymopoiesis and TrkC sensory neurons during dorsal root ganglia neurogenesis. It also mediates transforming growth factor-β signaling in dendritic cells and is essential for development of skin Langerhans cells. Here, we report that Runx3 is involved in the development of skin dendritic epidermal T cells (DETCs); an important component of tissue immunoregulation. In developing DETCs, Runx3 regulates expression of the αEβ7 integrin CD103, known to affect migration and epithelial retention of DETCs. It also regulates expression of IL-2 receptor β (IL-2Rβ) that mediates cell proliferation in response to IL-2 or IL-15. In the absence of Runx3, the reduction in CD103 and IL-2Rβ expression on Runx3 −/− DETC precursors resulted in impaired cell proliferation and maturation, leading to complete lack of skin DETCs in Runx3 −/− mice. The data demonstrate the requirement of Runx3 for DETCs development and underscore the importance of CD103 and IL-2Rβ in this process. Of note, while Runx3 −/− mice lack both DETCs and Langerhans cells, the two most important components of skin immune surveillance, the mice did not develop skin lesions under pathogen-free (SPF) conditions.

From the editors

From the editors

Beating Leishmania's iron armor

Leishmania parasites that lack an essential iron transporter lose the ability to replicate within macrophages. These parasites persist but do not lead to host pathology, report Chau Huynh et al. (page 2363). Targeting the newly identified LIT1 pathway could be a potential route to leishmaniasis therapy. Figure 1 Replication of Leishmania inside phagolysome (top) fails when LIT1 is deleted (bottom). Intracellular parasites need iron to grow and multiply, but within the phagolysosome compartment, which such parasites inhabit, the supply of iron is limited. Specialized iron transporters had been identified in other intracellular pathogens, but none had been found in Leishmania, until now. Huynh and colleagues searched the Leishmania genome for sequences with similarity to known iron transporters and found a novel gene they call Leishmania iron transporter 1 (LIT1). Having confirmed that LIT1 behaved as an iron transporter in yeast, the team deleted the gene from Leishmania to see how the bugs would fare in mouse macrophages. By 48 to 72 hours, the number of wild-type Leishmania per phagolysome compartment had significantly increased. LIT1−/− Leishmania, on the other hand, failed to multiply. Their phagolysosome homes appeared smaller, and the bugs themselves showed signs of degeneration. Whether these bugs would be eventually cleared from the cells is unknown, as macrophages do not survive well in culture beyond 72 hours. Mice infected with wild-type Leishmania developed skin lesions indicative of cutaneous leishmaniasis. However, when infected with LIT1−/− Leishmania, no lesions appeared. Interestingly, samples from the mice injected with LIT1−/− bugs still contained live Leishmania up to 6 months later, despite the pathology-free status of these mice. Removal of the iron transporter LIT1, thus, obliterates Leishmania's virulence and renders the bug incapable of replication, yet does not kill it. The team is currently interested in determining the intracellular location of the surviving LIT1−/− bugs and in seeing whether this location allows them to acquire iron by an alternative route. This is an important question—if answered, it could lead to new treatments capable of completely eliminating the parasites from their infected hosts.

Mice with a Targeted Mutation of Patched2 Are Viable but Develop Alopecia and Epidermal Hyperplasia

Hedgehog (Hh) signaling plays pivotal roles in tissue patterning and development in Drosophila melanogaster and vertebrates. The Patched1 (Ptc1) gene, encoding the Hh receptor, is mutated in nevoid basal cell carcinoma syndrome, a human genetic disorder associated with developmental abnormalities and increased incidences of basal cell carcinoma (BCC) and medulloblastoma (MB). Ptc1 mutations also occur in sporadic forms of BCC and MB. Mutational studies with mice have verified that Ptc1 is a tumor suppressor. We previously identified a second mammalian Patched gene, Ptc2, and demonstrated its distinct expression pattern during embryogenesis, suggesting a unique role in development. Most notably, Ptc2 is expressed in an overlapping pattern with Shh in the epidermal compartment of developing hair follicles and is highly expressed in the developing limb bud, cerebellum, and testis. Here, we describe the generation and phenotypic analysis of Ptc2(tm1/tm1) mice. Our molecular analysis suggests that Ptc2(tm1) likely represents a hypomorphic allele. Despite the dynamic expression of Ptc2 during embryogenesis, Ptc2(tm1/tm1) mice are viable, fertile, and apparently normal. Interestingly, adult Ptc2(tm1/tm1) male animals develop skin lesions consisting of alopecia, ulceration, and epidermal hyperplasia. While functional compensation by Ptc1 might account for the lack of a strong mutant phenotype in Ptc2-deficient mice, our results suggest that normal Ptc2 function is required for adult skin homeostasis.

Arsenic Exposure and Age- and Sex-Specific Risk for Skin Lesions: A Population-Based Case–Referent Study in Bangladesh

BackgroundThe objective of this population-based case–referent study in Matlab, Bangladesh, was to assess the susceptibility to arsenic-induced skin lesions by age and sex, in a population drinking water from As-contaminated tube wells.MethodsIdentification of As-related skin lesions was carried out in three steps: a) screening of the entire population > 4 years of age (n = 166,934) by trained field teams; b) diagnosis of suspected As-related cases by physicians; and c) confirmation by experts based on physicians’ records and photographs. A total of 504 cases with skin lesions were confirmed. We randomly selected 2,201 referents from the Matlab health and demographic surveillance system; 1,955 were eligible, and 1,830 (94%) were available for participation in the study. Individual history of As exposure was based on information obtained during interviews and included all drinking-water sources used since 1970 and concentrations of As (assessed by atomic absorption spectrophotometry) in all the tube wells used.ResultsCases had been exposed to As more than referents (average exposure since 1970: male cases, 200 μg/L; female cases, 211 μg/L; male referents, 143 μg/L; female referents, 155 μg/L). We found a dose–response relationship for both sexes (p < 0.001) and increased risk with increasing socioeconomic status. Males had a higher risk of obtaining skin lesions than females (odds ratio 10.9 vs. 5.78) in the highest average exposure quintile (p = 0.005). Start of As exposure (cumulative exposure) before 1 year of age was not associated with higher risk of obtaining skin lesions compared to start of As exposure later in life.ConclusionsThe results demonstrate that males are more susceptible than females to develop skin lesions when exposed to As in water from tube wells.

Drug Hypersensitivity Syndrome Caused by Minocycline

Minocycline is a commonly prescribed drug for the treatment of acne. Its use is generally not associated with systemic side effects. To describe a case of minocycline-induced drug hypersensitivity syndrome in a 20-year-old Japanese woman. Following 2 months of minocycline treatment, the patient developed skin lesions composed of exudative maculopapules, purpuratous macules, and target-like, erythema multiforme-like plaques over most of her body. In addition, she had fever, abnormal liver function tests, eosinophilia, and atypical lymphocytosis. Laboratory tests indicated no elevation of antibody titers against cytomegalovirus, Epstein-Barr virus, and human herpesvirus 6. Her ongoing exposure to minocycline was stopped, and treatment with oral prednisolone was begun. Her signs, symptoms, and laboratory abnormalities then began to resolve. Subsequently, the syndrome was observed to return briefly in response to an oral challenge with minocycline. Minocycline is able to elicit a drug hypersensitivity syndrome that can resemble infectious mononucleosis. This drug reaction can be treated effectively by cessation of exposure to this drug and steroid therapy.

CD-3 positive extranodal T-cell lymphoma of nasal type with skin involvement

A 40-year-old previously healthy lady presented with nasal obstruction and localized plaques over the right arm. She developed complete nasal obstruction due to a mass in the right nasal cavity and skin lesions that ulcerated to present as ecthyma gangrenosum like lesions. Patient's condition deteriorated fast and she developed icterus with fatal outcome within 4 weeks of developing skin lesions. Nasal and skin biopsy revealed angiocentric T-cell lymphoma, which on immuno-phenotyping revealed CD-3 positive; and CD-20, CD-30, ALK and EMA negativity. She was seronegative for HIV. Final diagnosis of CD-3 positive extranodal T-cell lymphoma of nasal type was made. Extranodal T-cell lymphomas are very aggressive NHLs with poor prognosis. Prognosis depends on histology, stage of the disease and sites of involvement. NK/T cell lymphoma of nasal type is common with EBV association. Skin involvement is rare and is also an indicator of poor prognosis.

Dietary phytosterols reduce probucol-induced atherogenesis in apo E-KO mice

We have previously shown strong pro-atherogenic effects of probucol in apolipoprotein E-knockout (apo E-KO) mice. The aims of the present study were to investigate whether (a) dietary phytosterols reduce probucol-induced atherogenesis and (b) beneficial interactions exist between these agents. Male apo E-KO mice fed with an atherogenic diet supplemented with phytosterols or probucol or their combination for 14 weeks. Single therapy with either phytosterols or probucol resulted in a 25% reduction in plasma total cholesterol (TC) concentrations as compared to the control group. The effects of the combination therapy were more profound (60% reduction). While phytosterols reduced atherogenesis by 60%, probucol caused an increase of 150% in atherogenesis. Addition of phytosterols to probucol substantially reduced pro-atherogenic effects of probucol. This was associated with improved high density lipoprotein (HDL) concentrations. The ratio of TC to HDL cholesterol was markedly reduced in the combination therapy group as compared to the probucol-treated group. A strong positive association between the ratio of TC to HDL cholesterol and the extent of atherosclerotic lesions was observed. The coronary arteries of the probucol-treated group showed various stages of atherogenesis from infiltration of monocytes into intima to complete occlusion of the vessel by atheromatous lesions. Such pathological findings were not observed in the combination therapy group. Approximately 40% of the mice in the probucol-treated group and 10% of the animals in the combination therapy group developed skin lesions. Further studies warrant the investigation of the underlying mechanisms of the observed beneficial interactions between dietary phytosterols and probucol.

Mycotic dermatitis with digital gangrene and osteomyelitis, and protozoal intestinal parasitism in Marlborough green geckos (Naultinus manukanus)

CASE HISTORY: Thirty adult Marlborough green geckos (Naultinus manukanus) were collected from Stephens Island and held over winter, prior to their translocation. Five adult geckos developed skin lesions after husbandry changes affected the humidity of their enclosures. Two geckos underwent ecdysis and recovered. One animal died and two others progressively worsened and were presented for treatment. CLINICAL AND PATHOLOGICAL FINDINGS: The geckos were in poor body condition and had multiple black powdery lesions and solitary raised white nodules on their skin. Both geckos died despite topical and supportive treatment. Histopathology showed the skin nodules contained branching non-septate hyphae infiltrating necrotic epidermal tissue, and associated dermal inflammation. There was necrosis of several digits and mycotic osteomyelitis. Mucor ramosissimus was cultured from skin biopsies from each animal. Large numbers of motile protozoa, resembling Trichomonas, and another unidentifiable, were recovered from fresh faecal smears, and Nyctotherus sp protozoa were present in the lumen of the intestine of one animal post mortem. DIAGNOSIS: Mycotic dermatitis with digital gangrene and osteomyelitis due to Mucor ramosissimus, and enteric protozoal parasitism with Trichomonas sp and Nyctotherus sp. CLINICAL RELEVANCE: The clinical course and pathological findings of mycotic dermatitis in two Marlborough green geckos involved in a wildlife translocation in New Zealand are reported, and also the first record of the Marlborough green gecko as a host for the enteric protozoa Trichomonas sp and Nyctotherus sp.

The impact of diet and betel nut use on skin lesions associated with drinking-water arsenic in Pabna, Bangladesh.

An established exposure–response relationship exists between water arsenic levels and skin lesions. Results of previous studies with limited historical exposure data, and laboratory animal studies suggest that diet may modify arsenic metabolism and toxicity. In this study, we evaluated the effect of diet on the risk of arsenic-related skin lesions in Pabna, Bangladesh. Six hundred cases and 600 controls loosely matched on age and sex were enrolled at Dhaka Community Hospital, Bangladesh, in 2001–2002. Diet, demographic data, and water samples were collected. Water samples were analyzed for arsenic using inductively coupled plasma mass spectroscopy. Betel nut use was associated with a greater risk of skin lesions in a multivariate model [odds ratio (OR) = 1.67; 95% confidence interval (CI), 1.18–2.36]. Modest decreases in risk of skin lesions were associated with fruit intake 1–3 times/month (OR = 0.68; 95%CI, 0.51–0.89) and canned goods at least 1 time/month (OR = 0.41; 95% CI, 0.20–0.86). Bean intake at least 1 time/day (OR = 1.89; 95% CI, 1.11–3.22) was associated with increased odds of skin lesions. Betel nut use appears to be associated with increased risk of developing skin lesions in Bangladesh. Increased intake of fruit and canned goods may be associated with reduced risk of lesions. Increased intake of beans may be associated with an increased risk of skin lesions. The results of this study do not provide clear support for a protective effect of vegetable and overall protein consumption against the development of skin lesions, but a modest benefit cannot be excluded.

The detection of lumpy skin disease virus in samples of experimentally infected cattle using different diagnostic techniques

Lumpy skin disease (LSD) is a disease of cattle, primarily in Africa and Madagascar and rarely in the Middle East. It is caused by a capripoxvirus that belongs to the family Poxviridae. The disease is of economic importance in endemic areas. Effective control of LSD requires accurate and rapid laboratory techniques to confirm a tentative clinical diagnosis. Comparative studies on different diagnostic tests used at different stages of the disease have not been done. The aim of this study was to compare several of these tests. Six seronegative bulls, between 11 and 20 months of age, were infected intravenously and kept in an insect-free facility. The course of the infection was monitored. During a 3-month period blood samples and skin biopsies were collected for virus isolation and polymerase chain reaction (PCR). Skin biopsies were also examined using transmission electron microscopy (TEM). The incubation period in infected animals varied from 4-5 days. The length of the viraemic period did not correlate with the severity of clinical disease. Viraemia was detected from 1-12 days using virus isolation and from 4-11 days using the PCR, which is longer than has previously been reported. Virus was isolated from skin biopsies until Day 39 post infection (p.i.) and PCR could demonstrate viral DNA until Day 92 p.i. Transmission electron microscopy of negatively stained skin biopsies detected LSD virus only in one of the four bulls that developed skin lesions until Day 33 p.i. The PCR was a fast and sensitive method to demonstrate viral DNA in blood and skin samples. It could detect viral nucleic acid in skin lesions 53 days longer than virus isolation. Virus isolation from blood and skin samples was sensitive and reliable, but as a single test it may be too time-consuming to use although this depends on how rapidly the diagnosis must be confirmed. In conclusion, this study showed the PCR to be superior in detecting LSD virus from blood and skin samples. However, virus isolation is still required when the infectivity of the LSD virus is to be determined.

Decrements in Lung Function Related to Arsenic in Drinking Water in West Bengal, India

During 1998-2000, the authors investigated relations between lung function, respiratory symptoms, and arsenic in drinking water among 287 study participants, including 132 with arsenic-caused skin lesions, in West Bengal, India. The source population involved 7,683 participants who had been surveyed for arsenic-related skin lesions in 1995-1996. Respiratory symptoms were increased among men with arsenic-caused skin lesions (versus those without lesions), particularly "shortness of breath at night" (odds ratio (OR) = 2.8, 95% confidence interval (CI): 1.1, 7.6) and "morning cough" (OR = 2.8, 95% CI: 1.2, 6.6) in smokers and "shortness of breath ever" (OR = 3.8, 95% CI: 0.7, 20.6) in nonsmokers. Among men with skin lesions, the average adjusted forced expiratory volume in 1 second (FEV1) was reduced by 256.2 ml (95% CI: 113.9, 398.4; p < 0.001) and the average adjusted forced vital capacity (FVC) was reduced by 287.8 ml (95% CI: 134.9, 440.8; p < 0.001). In men, a 100-microg/liter increase in arsenic level was associated with a 45.0-ml decrease (95% CI: 6.2, 83.9) in FEV1 (p = 0.02) and a 41.4-ml decrease (95% CI: -0.7, 83.5) in FVC (p = 0.054). Women had lower risks than men of developing skin lesions and showed little evidence of respiratory effects. In this study, consumption of arsenic-contaminated water was associated with respiratory symptoms and reduced lung function in men, especially among those with arsenic-related skin lesions.

DRINKING WATER EXPOSURE TO ARSENIC, POLYMORPHISMS IN GSTT1, GSTM1 AND GSTP1 AND METHYLATION CAPACITY

ISEE-291 Introduction: Individual differences in the biotransformation of arsenic have has been hypothesized to be a factor in arsenic related disease. Recently there has been a suggestion that the metabolism and methylation of arsenic is a toxification pathway rather than a detoxification pathway. The methylated metabolites of arsenic are thought to be more toxic than originally categorized. Aim: We examined whether polymorphisms in GSTT1, GSTP1 and GSTM1 were associated with primary (MMA monomethylarsonic acid / As III arsenite +As V arsenate) and secondary methylation capacity (DMA dimethylarsinic acid /MMA monomethylarsonic acid). We also examined the association between primary and secondary methylation capacity and development of arsenic related skin lesions and whether this relationship was modified by GSTT1, GSTM1, and GSTP1 polymorphisms. Methods: 592 cases and 597 controls, frequency matched on age and gender, were enrolled at Dhaka Community Hospital, Bangladesh in 2001-2002. Demographic data, toenail and water samples were collected for all subjects. Trained physicians diagnosed skin lesions. Nail and water arsenic levels were determined using ICP-MS EPA Method 200.8. Genotyping was done with multiplex PCR and TaqMan. Statistical analysis was done in SAS version 8.2 and plots in R. Results: While drinking water concentration of arsenic was related to secondary methylation capacity and individual metabolites MMA and DMA, it was not related to primary methylation capacity. Polymorphisms in GSTT1, GSTM1 and GSTP1 did not modify primary or secondary methylation capacity. Betel nut use was significantly associated with secondary methylation capacity. Individual metabolites, DMA and MMA, are associated with arsenic concentration of drinking water; however not associated with the polymorphisms. The odds of developing skin lesions is associated with an increase of secondary methylation capacity, but not primary methylation capacity. Conclusions: GSTT1, GSTM1 and GSTP1 polymorphisms do not impact primary or secondary methylation capacity in the metabolism of arsenic.