Genome-wide association study of dermatophytosis in the UK Biobank cohort.

BACKGROUND: Analyses of the hereditary propensity to dermatophytosis have revealed several proven genetic relationships. They include CARD9 deficiency, HLA-DR4 and HLA-DR8 type and genes encoding interleukin-22, defensin 2 and 4, and genetic defects in dectin-1, which increased the prevalence of dermatophytosis in families and were involved in the inheritance of susceptibility in their members. METHODS: To further investigate the genetic basis of dermatophytosis, we performed a genome-wide association study (GWAS) of the UK Biobank cohort. To identify cases of dermatophytosis, we used ICD10 code B35, which covers Tinea barbae, Tinea capitis, Tinea unguium, Tinea manuum, Tinea pedis, Tinea corporis, Tinea imbricata, Tinea cruris, other dermatophytoses and dermatophytosis, unspecified. Data processing was performed on Minerva, a Linux mainframe with Centos 7.6, at the Icahn School of Medicine at Mount Sinai. We used PLINK, a whole-genome association analysis toolset, to analyse the UKB chromosome files and the UK Biobank Data Parser (ukbb parser), a python-based package that allows easy interfacing with the large UK Biobank dataset. We used LocusZoom for the Manhattan and q-q plots. Other statistical analyses were done with R and SPSS 25. RESULTS: Genome-wide association study (GWAS) and meta-analysis association statistics highlighted one susceptibility locus, Tubulointerstitial Nephritis Antigen (TINAG), with genome-wide significance for dermatophytosis. The top SNP was rs16885197, a missense variant within TINAG, position chr6:54308557, alleles A > G, minor allele frequency (MAF) 0.014. Multivariate logistic regression indicated that the minor G allele increased odds ratio of dermatophytosis by 7.8. Carrying two G alleles raised dermatophytosis odds ratio by a factor of 14. CONCLUSION: More research into genetic and other predisposing factors for dermatophytosis is critical because of the implications for prophylaxis and therapy. It might be possible to prevent infection and recurrence by identifying people who are vulnerable to chronic dermatophytosis. Identifying high-risk families would enable their members to be educated about the dangers of fungal diseases. New therapeutic techniques to target altered hormonal and immune response pathways might be created. TINAG is a prospective target that should be investigated, based on the findings of this article.

Cannabis smoking and age-related macular degeneration in the UK Biobank cohort.

BACKGROUND: A major cause of blindness in older persons is age-related macular degeneration (AMD). Cigarette smoking is one of the major risk factors for AMD. In the present study, we analyzed UK Biobank data to determine whether smoking cannabis, like cigarettes, might be related to AMD. METHODS: Our UK Biobank application was approved as UKB project 57245 (S.L., P.H.R.). Our analysis included all subjects with AMD and cannabis smoking information. The diagnosis of AMD was ascertained based on the 10th Revision of the International Classification of Diseases (ICD10), H35.3. Age at time of diagnosis of AMD was obtained from data field 5923. Cannabis information was recorded in UKB category 143, data field 20453, ever taken cannabis. A touch screen posed the question, "Have you taken CANNABIS (cannabis, grass, hash, ganja, blow, draw, skunk, weed, spliff, dope), even if it was a long time ago?" Possible answers were no, yes, 1-2 times, yes 3-10 times, yes, 11-100 times, yes, more than 100 times. RESULTS: Subjects who had used marijuana more than 100 times had a significantly reduced risk of AMD, compared to subjects who had never used marijuana, and use of marijuana every day was associated with less AMD than use of marijuana less than once a month. But subjects who used cannabis 100 times or more were significantly younger (8years) when they developed AMD than subjects who never used cannabis. CONCLUSION: Drusen, deposits of lipids, proteins, and cellular debris, accumulate in Bruch's membrane, limiting transport between the retinal pigment epithelium and the vasculature, triggering an inflammatory reaction. Marijuana can retard the inflammatory process because it is a powerful anti-inflammatory agent. Therefore, marijuana could reduce the risk of AMD. At the same time, blood vessels in the choriocapillaris below Bruch's membrane become more sparse with age. This phenomenon is believed to be a starting point for AMD. Marijuana can accelerate the loss of blood vessels due to its anti-angiogenic properties. Therefore, marijuana use might cause AMD to develop sooner in younger people.

Cannabis smoking and glaucoma in the UK Biobank cohort.

BACKGROUND: Cigarette smoking is a well-known risk factor for primary open-angle glaucoma (POAG). Tobacco smoke contains a variety of chemicals, including nicotine, free radicals, and carbon monoxide, all of which can play a role in the development of POAG. Cannabis smoke, like tobacco smoke, contains a comparable variety of carcinogenic and toxic compounds. In the present study, we analyzed UK Biobank data to determine whether smoking cannabis, like cigarettes, might be related to POAG. METHODS: Our analysis included all subjects with glaucoma and cannabis smoking information. Data processing was performed on Minerva, a Linux mainframe with Centos 7.6, at the Icahn School of Medicine at Mount Sinai. We used the UK Biobank Data Parser (ukbb parser), a python-based package that allows easy interfacing with the large UK Biobank dataset. Statistical analysis was performed with SPSS 25 and R. RESULTS: Subjects who used cannabis 100 times or more were significantly younger (10years) when they developed glaucoma than subjects who never used cannabis. The effects of age (P<0.001) and cigarettes (P=0.014) on POAG incidence were significant, but the effect of cannabis use was not (P=0.662). The effect of cannabis use on age at glaucoma diagnosis was significant and unrelated to the effects on intraocular pressure or cigarette pack-years smoked. Analysis of intraocular pressure showed that mean pressures of subjects who used cannabis 11-100 times were significantly lower than those who took no cannabis. CONCLUSION: Inhaling cannabis smoke is injurious to the eye, but in the case of glaucoma the manifestations are different from those of inhaled tobacco smoke. Cannabis reduces intraocular pressure but accelerates glaucoma development. Cannabis does not increase risk of glaucoma. Cigarette smoking increases risk of glaucoma and significantly accelerates glaucoma development. Apparently, early use of cannabis leads to the earlier development of glaucoma.

Marijuana smoking and cataract.

BACKGROUND: Cigarette smoking is a well-known risk factor for cataract and other ailments, including heart disease, lung cancer, and chronic obstructive pulmonary disease. Cannabis smoke, like tobacco smoke, contains a comparable variety of carcinogenic and toxic compounds. PURPOSE: In the present study, we analyzed UK Biobank data to determine whether smoking cannabis, like cigarettes, might be related to cataract. METHODS: Our analysis included all UK Biobank subjects with cataracts and information on cannabis and cigarette smoking habits. The diagnosis of cataract was ascertained using the 10th Revision of the International Classification of Diseases (ICD10), H25. The age at diagnosis of cataract was obtained from UK Biobank data field 4700. Cannabis information was recorded in UK Biobank category 143, data field 20453, ever taken cannabis. RESULTS: Subjects who used cannabis 11-100 times or more were significantly younger (4-5 years) when they developed cataract than subjects who never used cannabis. To determine the relationship of current cigarette smoking to cannabis use and age at cataract, the univariate general linear model of SPSS was used, dependent variable age at cataract, fixed factor cannabis use, random factor pack years of cigarettes smoked. Cannabis use was significantly related to age at cataract diagnosis (P<0.001) and independent of the effect of pack-years of cigarettes smoked (P=0.008). Linear regression revealed an insignificant relationship between pack-years of cigarette smoking and age at cataract diagnosis (P=0.073). To further evaluate the relationship of cannabis to cataract, propensity score matching was performed. We identified 28,432 subjects with cataract. Current cigarette smoking and age were covariates; cannabis use (yes/no) was the indicator variable. Current cigarette smoking was significantly associated with a 1.2 odds ratio for cataract. Cannabis use was not significantly associated with the odds ratio for cataract. CONCLUSION: Like tobacco smoke, cannabis smoke contains thousands of organic and inorganic chemical compounds. Cannabis tar is chemically similar to tar found in tobacco smoke, and over fifty known carcinogens have been identified in cannabis smoke, including nitrosamines, reactive aldehydes, and polycyclic hydrocarbons. Thus, the association of cannabis with cataract that we report here is not entirely surprising. Further studies are warranted.

Diabetes, cigarette smoking and transcription factor 7-like 2 (Tcf7L2) in the UK Biobank cohort.

Background ­: Smokers are 30 to 40 percent more likely to develop type 2 diabetes than non-smokers. A type 2 diabetes gene, Tcf7L2, which had lost activity, caused rats to consume more nicotine. In the present study, we used data from the UK Biobank to examine the relationship of smoking, type 2 diabetes, and Tcf7L2 in human subjects. Methods ­: The Tcf7L2 gene has two SNPs, rs7903146 and rs4506565, reported to be associated with type 2 diabetes. They have approximately equal power to estimate risk for type 2 diabetes, and the results from one correlate 92% with the other. We examined the genotypes of these SNPs and cigarette consumption. Results ­: Genotype TT, linked to type 2 diabetes, smoked the least. But because of the large sample size (approximately 111,000 subjects) the tiny difference in cigarettes smoked daily by each genotype group (effect size), while statistically significant, is probably clinically meaningless. The average subject smoked 19 cigarettes daily, with a difference of 0.12 cigarette between each genotype group. Conclusion ­: The fact that Tcf7L2 is involved in nicotine addiction in rats but not in humans, as UKBB data suggest, is hardly surprising. Humans and rodents descended from a common ancestor about 80 million years ago, with rats and mice diverging between 12 and 24 million years ago. Thus, over millions of years Tcf7L2 may have developed vastly different functions in rodents and humans. Genome Wide Association Studies have revealed at least 65 different loci linked to type 2 diabetes. Genes associated with type 2 diabetes include Tcf7L2, PPARG, FTO, KCNJ11, NOTCH2, WFS1, IGF2BP2, SLC30A8, JAZF1, HHEX, DGKB, CDKN2A, CDKN2B, KCNQ1, HNF1A, HNF1B MC4R, GIPR, HNF4A, MTNR1B, PARG6, ZBED3, SLC30A8, CDKAL1, GLIS3, GCK, GCKR, among others. Perhaps one or more of these genes might be the intermediary between type 2 diabetes and cigarette smoking. Further studies are warranted.

Contexte ­: Les fumeurs sont 30 à 40% plus susceptibles de développer un diabète de type 2 que les non-fumeurs. Un gène du diabète de type 2, Tcf7L2, qui avait perdu son activité, a poussé les rats à consommer plus de nicotine. Dans la présente étude, nous avons utilisé les données de la UK Biobank pour examiner la relation entre le tabagisme, le diabète de type 2 et le Tcf7L2 chez des sujets humains. Méthodes ­: Le gène Tcf7L2 possède deux SNPS, rs7903146 et rs4506565, qui seraient associés au diabète de type 2. Ils ont à peu près la même puissance pour estimer le risque de diabète de type 2, et les résultats de l'un sont en corrélation à 92% avec l'autre. Nous avons examiné les génotypes de ces SNP et la consommation de cigarettes. Résultats ­: Le génotype TT, lié au diabète de type 2, fumait le moins. Mais en raison de la grande taille de l'échantillon (environ 111 000 sujets), la petite différence de cigarettes fumées quotidiennement par chaque groupe de génotypes (taille de l'effet), bien que statistiquement significative, est probablement sans signification clinique. Le sujet moyen fumait 19 cigarettes par jour, avec une différence de 0,12 cigarette entre chaque groupe de génotype. Conclusion ­: Le fait que le Tcf7L2 soit impliqué dans la dépendance à la nicotine chez le rat mais pas chez l'homme, comme le suggèrent les données de l'UKBB, n'est guère surprenant. Les humains et les rongeurs descendent d'un ancêtre commun il y a environ 80 millions d'années, les rats et les souris divergeant il y a entre 12 et 24 millions d'années. Ainsi, au cours de millions d'années, Tcf7L2 peut avoir développé des fonctions très différentes chez les rongeurs et les humains. Des études sur l'association à l'échelle du génome ont révélé au moins 65 loci différents liés au diabète de type 2. Les gènes associés au diabète de type 2 comprennent Tcf7L2, PPARG, FTO, KCNJ11, NOTCH2, WFS1, IGF2BP2, SLC30A8, JAZF1, HHEX, DGKB, CDKN2A, CDKN2B, KCNQ1, HNF1A, HNF1B MC4R, GIPR, HNF4A, MTGNR1B, ZNF4A, MTGNR1B, SLC30A8, CDKAL1, GLIS3, GCK, GCKR, entre autres. Peut-être qu'un ou plusieurs de ces gènes pourraient être l'intermédiaire entre le diabète de type 2 et le tabagisme. D'autres études sont justifiées.

Shellfish allergy.

Seafood plays an important role in human nutrition and health. The growing international trade in seafood species and products has added to the popularity and frequency of consumption of a variety of seafood products across many countries. This increased production and consumption of seafood has been accompanied by more frequent reports of adverse health problems among consumers as well as processors of seafood. Adverse reactions to seafood are often generated by contaminants but can also be mediated by the immune system and cause allergies. These reactions can result from exposure to the seafood itself or various non-seafood components in the product. Non-immunological reactions to seafood can be triggered by contaminants such as parasites, bacteria, viruses, marine toxins and biogenic amines. Ingredients added during processing and canning of seafood can also cause adverse reactions. Importantly all these substances are able to trigger symptoms which are similar to true allergic reactions, which are mediated by antibodies produced by the immune system against specific allergens. Allergic reactions to 'shellfish', which comprises the groups of crustaceans and molluscs, can generate clinical symptoms ranging from mild urticaria and oral allergy syndrome to life-threatening anaphylactic reactions. The prevalence of crustacean allergy seems to vary largely between geographical locations, most probably as a result of the availability of seafood. The major shellfish allergen is tropomyosin, although other allergens may play an important part in allergenicity such as arginine kinase and myosin light chain. Current observations regard tropomyosin to be the major allergen responsible for molecular and clinical cross-reactivity between crustaceans and molluscs, but also to other inhaled invertebrates such as house dust mites and insects. Future research on the molecular structure of tropomyosins with a focus on the immunological and particularly clinical cross-reactivity will improve diagnosis and management of this potentially life-threatening allergy and is essential for future immunotherapy.

Occupational asthma and allergy in snow crab processing in Newfoundland and Labrador.

BACKGROUND: Risk factors and prevalence of occupational asthma (OA) and occupational allergy (OAl) in the snow crab-processing industry have been poorly studied. OBJECTIVE: To estimate the prevalence of OA and OAl in snow crab-processing workers and determine their relationship with exposure to snow crab allergens and other potential risk factors. METHODS: A total of 215 workers (120 female/95 male) were recruited from four plants in Newfoundland and Labrador, Canada in 2001-2002. Results from questionnaires, skin-prick tests to snow crab meat and cooking water, specific IgEs against the latter, spirometry and peak flow monitoring were used to develop a diagnostic algorithm. An index based on work history and exposure measurements of snow crab aeroallergens was developed to estimate the cumulative exposure for each worker. RESULTS: The prevalences of almost certain or highly probable OA and OAl were 15.8% and 14.9%, respectively. A high cumulative exposure to crab allergens, in jobs mostly held by women, was associated with OA (odds ratio (OR) = 14.0, 95% CI 3.0 to 65.8) (highest vs lowest Cumulative Exposure Index) and with OAl (OR = 7.1, 95% CI 1.9 to 29.0); job held when symptoms started (cleaning, packing, freezing) also predicted OA (OR = 3.9, 95% CI 1.6 to 8.7) and OAl (OR = 3.2, 95% CI 1.4 to 7.5). Atopy (OR = 2.8, 95% CI 1.2 to 6.8), female gender (OR = 10.7, 95% CI 3.6 to 32.1) and smoking were significant determinants for OA (OR = 3.1, 95% CI 1.3 to 7.4). CONCLUSIONS: The prevalences of OA and OAl are high in snow crab-processing workers of Canada's East Coast. Cumulative exposure to snow crab allergens was related to the prevalences of OA and OAl in a dose-response manner taking into account atopy, gender and smoking.

Do shrimp-allergic individuals tolerate shrimp-derived glucosamine?

BACKGROUND: There is concern that shrimp-allergic individuals may react to glucosamine-containing products as shrimp shells are a major source of glucosamine used for human consumption. OBJECTIVE: The purpose of this study was to determine whether shrimp-allergic individuals can tolerate therapeutic doses of glucosamine. METHODS: Subjects with a history of shrimp allergy were recruited and tested for both shrimp reactivity via a prick skin test and shrimp-specific IgE by an ImmunoCAP assay. Fifteen subjects with positive skin tests to shrimp and an ImmunoCAP class level of two or greater were selected for a double-blind placebo-controlled food challenge (DBPCFC) using glucosamine-chondroitin tablets containing 1,500 mg of synthetically produced (control) or shrimp-derived glucosamine. Immediate reactions, including changes in peak flow and blood pressure, and delayed reactions (up to 24 h post-challenge) via questionnaire were noted and assessed. RESULTS: All subjects tolerated 1,500 mg of both shrimp-derived or synthetic glucosamine without incident of an immediate hypersensitivity response. Peak flows and blood pressures remained constant, and no subject had symptoms of a delayed reaction 24 h later. CONCLUSION: This study demonstrates that glucosamine supplements from specific manufacturers do not contain clinically relevant levels of shrimp allergen and therefore appear to pose no threat to shrimp-allergic individuals.

Structural, immunological and functional properties of natural recombinant Pen a 1, the major allergen of Brown Shrimp, Penaeus aztecus.

BACKGROUND: Recombinant allergens are considered the basis for new diagnostic approaches and development of novel strategies of allergen-specific immunotherapy. As Pen a 1 from brown shrimp Penaeus aztecus is the only major allergen of shrimp and binds up to 75% of all shrimp-specific IgE antibodies this molecule may be an excellent model for the usage of allergens with reduced IgE antibody-binding capacity for specific immunotherapy. AIM: The aim was to clone, express and characterize a full-length recombinant Pen a 1 molecule and compare it with natural Pen a 1 in regard to structural and immunological parameters such as IgE antibody capacity and ability to induce IgE-mediated mediator release. METHODS: Total RNA was isolated from P. aztecus and a rapid amplification of cDNA ends (5' RACE) was performed to obtain full-length cDNA coding for Pen a 1. Using a gene-specific primer, PCR was performed and full-length cDNA was cloned and sequenced. Recombinant His-tagged Pen a 1 was isolated from Escherichia coli under native conditions by immobilized metal affinity chromatography. Secondary structure of natural and recombinant Pen a 1 was compared by circular dichroism (CD) spectroscopy, and the IgE antibody-binding capacity evaluated by RAST. The allergenic potency was tested by the capability of natural and recombinant Pen a 1 to induce mediator release in a murine and human in vitro model of IgE-mediated type I allergy. RESULTS: The deduced amino-acid sequence was 284 residues long and amino-acid sequence identities with allergenic and non-allergenic tropomyosins ranged from 80% to 99% and 51% to 58%, respectively. The analysis of the secondary structure of natural and recombinant Pen a 1 by CD spectroscopic analysis showed that both nPen a 1 and rPen a 1 had alpha-helical conformation that is typical for tropomyosin. The IgE antibody binding capacities of nPen a 1 and r Pen a1 were found to be essentially identical by RAST. The mediator release experiments using both wild-type and humanized rat basophilic leukaemia 30/25 cells showed that rPen a 1 and nPen a 1 induced a similar level of mast cell activation. CONCLUSIONS: Recombinant Pen a 1 and natural Pen a 1 are structurally and immunologically identical and rPen a 1 may be used as the basis for component-resolved diagnosis and the generation of modified shrimp tropomyosin for allergen-specific immunotherapy. The results of the animal studies indicate that C3H/HeJ mice that were sensitized with shrimp extract in combination with cholera toxin as adjuvant may be a suitable model to study shrimp allergy.

Risks of allergic reactions to biotech proteins in foods: perception and reality.

In recent years, significant attention has been paid to the use of biotechnology to improve the quality and quantity of the food supply due in part to the projected growth in the world population, plus limited options available for increasing the amount of land under cultivation. Alterations in the food supply induced by classical breeding and selection methods typically involve the movement of large portions of genomic DNA between different plant varieties to obtain the desired trait. This is in contrast to techniques of genetic engineering which allows the selection and transfers specific genes from one species to another. The primary allergy risk to consumers from genetically modified crops may be placed into one of three categories. The first represents the highest risk to the allergic consumer is the transfer of known allergen or cross-reacting allergen into a food crop. The second category, representing an intermediate risk to the consumer, is the potential for replacing the endogenous allergenicity of a genetically-modified crop. The last category involves expression of novel proteins that may become allergens in man and generally represents a relatively low risk to the consumer, although this possibility has received attention of late. In order to mitigate the three categories of potential allergy risk associated with biotech crops, all genes introduced into food crops undergo a series of tests designed to determine if the biotech protein exhibits properties of known food allergens. The result of this risk assessment process to date is that no biotech proteins in foods have been documented to cause allergic reactions. These results indicate that the current assessment process is robust, although as science of allergy and allergens evolves, new information and new technology should help further the assessment process for potential allergenicity.

Environmental exposure characterization of fish processing workers.

BACKGROUND: Aerosolization of seafood and subsequent inhalation, during processing is a potential high-risk activity for allergic respiratory disease. OBJECTIVES: To quantify total thoracic particulate, protein concentration and specific fish (pilchard, anchovy) antigen concentrations in fish processing plants; to determine the correlation between these exposure metrics; and to identify the major determinants of variability and the optimal grouping strategies for establishing dose-response relationships for fish antigen exposures. METHODS: Exposure assessments were conducted on randomly selected individuals within each of the identified 'exposure groups' (EGs) in two fish processing factories. Personal time-integrated sampling was conducted with a thoracic fraction sampler and analysed for particulate mass, total protein and specific fish antigens. Exposure metrics were developed on the basis of individually measured exposures and average levels of these personal samples within EGs. The main components of the exposure variability were determined using ANOVA techniques. RESULTS: A total of 198 full-shift personal aerosol samples were collected and analysed. Twenty-two percent of the samples were below the limit of detection (LOD) for pilchard and 23% for anchovy assays. Personal sampling revealed wide variations across EGs in arithmetic mean concentrations of thoracic particulate 0.61 mg m(-3) (range: LOD-11.3), total protein 0.89 microg m(-3) (LOD-11.5), pilchard antigen 150 ng m(-3) (LOD-15 973) and anchovy antigen 552 ng m(-3) (LOD-75 748) levels. The fishmeal loading and bagging sections of both plants showed consistently high thoracic particulate mass (0.811-2.714 mg m(-3)), total protein (0.185-1.855 microg m(-3)), pilchard antigen (538-3288 ng m(-3)) and anchovy antigen (1708-15 431 ng m(-3)). The a priori strategy that grouped workers according to EGs produced reasonably satisfactory summary exposure metric statistics. An alternative grouping strategy based on department revealed comparable elasticity (exposure contrast). While the correlation between the log-transformed thoracic particulate mass and fish antigen concentrations were generally modest (Pearson's r = 0.32-0.35, P < 0.001), a high correlation was found between pilchard and anchovy antigen concentrations (Pearson's r = 0.71, P < 0.001). Models using factory and department grouping strategies accounted for a significant portion of the variability (adjusted r(2) = 0.18, P = 0.043) in pilchard antigen levels. Grouping strategies using a combination of factory and department yielded the highest degree of elasticity for thoracic particulate (0.38) and pilchard antigen (0.42) levels. CONCLUSIONS: Workers involved in bony fish processing are at risk of inhaling aerosols containing pilchard and anchovy fish antigens. Antigen exposures are highest during fishmeal production and bagging. Grouping strategies based on department and factory may provide a more efficient approach than a priori classification of EGs for evaluating fish antigen exposures.

Serum interleukin-8 is elevated in men with prostate cancer and bone metastases.

Aalinkeel et al. (1) have recently reported that gene expression of angiogenic factors correlates with metastatic potential of prostate cancer cells. The rationale for the study of Aalinkeel et al. is that a variety of growth factors, among them interleukin-8 (il-8), can induce angiogenesis (2). Further, parathyroid hormone related peptide (PTHrP) acts to induce il-8 production in prostate cancer cells via an intracrine pathway independent of its classical nuclear localization sequence. This novel pathway could mediate the effects of PTHrP on the progression of prostate cancer (3). We measured il-8 in the serum of 39 men with biopsy-proven prostate cancer. Their average age was 69 +/- 9 (mean +/- SD). Serum il-8 was measured with an automated chemiluminometric high sensitivity il-8 protein assay (Immulite, Diagnostic Products Corporation, Los Angeles, CA). We noted a significant elevation of il-8 in men with bone metastases, diagnosed by Tc-99 MDP bone scan, when compared to men with localized disease (Figure 1). Aalinkeel et al. found that il-8 was significantly higher in the more metastatic PC-3 and DU-145 prostate cancer cell lines, when compared to the poorly metastatic LnCAP cells. The results of our study of il-8 in men with prostate cancer support the findings of Aalinkeel et al. Therefore, new anti-angiogenic therapies targeting specific genes controlling prostate tumor metastasis may be of benefit in treating prostate cancer.

Enhancement of murine IgE antibody detection by IgG removal.

RATIONALE: Although animal models for the study of allergic reactions are desirable, the use of mice has been hindered by the lack of sufficiently sensitive in vitro immunoglobulin epsilon (IgE) antibody assays. The aim of this study was to enhance IgE antibody measurements by immunoglobulin gamma (IgG) depletion. METHODS: Seven- to eight-week-old female mice of four strains (C3H/HeJ, CBA/J, C57Bl/6J, and Balb/c) were immunized (20 mice/group) with shrimp or peanut extracts using Al(OH)(3) as adjuvant. Following immunization, animals were sacrificed by exsanguination and the sera of each group pooled. Initial measurements of IgE antibody levels by enzyme-linked immunosorbent assay (ELISA) were relatively low; IgG and IgE reactivity patterns by immunoblot were similar. Thus, sera from shrimp or peanut immunized mice were depleted of IgG (absorbed 3-6 times with immobilized protein G) and then tested for IgE antibody to shrimp or peanut allergen. RESULTS: A 3- to 5-fold increase in IgE antibody reactivity as measured by ELISA was demonstrated when >80-90% of the IgG was removed. This increase in detection of allergen-specific IgE occurred in sera from all mouse strains and to all allergens tested. In addition, reactivity of IgE antibodies to peanut or shrimp allergens by immunoblot increased visually approximately 4- to 10-fold. CONCLUSIONS: These studies indicate that allergen-specific IgG antibodies, which may be in more than 100-fold excess to IgE antibodies, interferes with detection of allergen-specific IgE, probably by competitive binding to allergenic epitopes. Substantial depletion of IgG antibodies (>80%) result in a significant increase in the sensitivity of the antibody measurements.

Immunoglobulin E antibody reactivity to the major shrimp allergen, tropomyosin, in unexposed Orthodox Jews.

BACKGROUND: Assessment of allergic (IgE antibody-mediated) reactions to foods may become complicated by cross-reactivity that can occur among certain food families and between foods and seemingly unrelated allergens. OBJECTIVE: The allergenic properties of tropomyosin (muscle-derived protein) have been recently demonstrated in invertebrates such as cockroaches, dust mites, and shrimp. In view of a possible cross-reactivity between food allergens and related allergens from animal sources, we designed a study to assess IgE antibody reactivity to the major shrimp allergen, Pen a 1, in an unexposed population of Orthodox Jews, who observe Kosher dietary laws that prohibit eating shellfish. METHODS: Nine subjects, who reacted positively by skin tests to shrimp (Penaeus setiferous), were selected for the study. Subjects (two females, seven males) ranged in age from 14 to 32 years (mean 20.4). All subjects were strictly observant of Jewish tradition and had no prior exposure to seafood (regarded as a non-Kosher food). Serum was obtained from all the subjects and tested for IgE antibody reactivity to shrimp and dust mite. RESULTS: All subjects reported symptoms of perennial allergic rhinitis, five had history of asthma, atopic dermatitis, and/or sinusitis. All had positive skin prick tests to shrimp and house dust mite (HDM) (Dermatophagoides farinae, D. pteronyssinus, or both); 2/7 subjects were positive to cockroach mix (Blattella germanica and Periplaneta americana). Sera of 4/9 subjects demonstrated specific IgE antibodies by RAST to shrimp (7.0-20.0%), 3/9 to Pen a 1 (6.3-24.1%), and 3/9 to shrimp or Pen a 1 by immunoblot. IgE binding to Pen a 1 was inhibited with either mite or cockroach extracts as demonstrated by RAST and/or immunoblot inhibition analysis. CONCLUSIONS: These studies indicate that IgE antibody reactivity to a major food allergen, shrimp, can occur in an unexposed population of individuals; some subjects allergic to HDM and/or cockroach show substantial IgE antibody reactivity to the major shrimp allergen Pen a 1 (tropomyosin). Based on inhibition with cockroach and/or dust mite extracts, this reactivity appears to be due to cross-reacting tropomyosins.

Putative protein markers in the sera of men with prostatic neoplasms.

OBJECTIVE: To describe the preliminary identification of serum proteins that may be diagnostic markers in prostate cancer. PATIENTS AND METHODS: The study included 11 men referred for treatment of localized prostate cancer, 12 with benign prostatic hyperplasia (BPH) and 12 disease-free controls. For serum protein analysis, the protein-chip array surface-enhanced laser desorption/ionization (SELDI) technique was used (Ciphergen Biosystems, Fremont, CA). SELDI combines protein-chip technology with time-of-flight mass spectrometry, and offers the advantages of speed, simplicity and sensitivity. RESULTS: Three protein peaks were identified in the serum of men with prostate cancer and BPH, but not in controls, with relative molecular masses of 15.2, 15.9 and 17.5 kDa. These three proteins were significantly associated with BPH and prostate cancer when compared with controls (P = 0.001, 0.004, and 0.011, respectively, Kruskal-Wallis test). Interestingly, the 17.5 kDa protein was more abundant in five men with stage T1 prostate cancer than in eight with stage T2 (P = 0.016, two tailed Mann-Whitney U-test corrected for ties). CONCLUSIONS: These proteins, particularly the 15.9 kDa one, may be used for the diagnosis or monitoring of prostate cancer and differentiation from BPH, and have the potential for antibody-based chip SELDI-TOF technology. Identified proteins may be targets for immunotherapy.

Seafood allergy and allergens: a review.

Seafoods are composed of diverse sea organisms and humans are allergic to many of them. Tropomyosin is a major allergen in many shellfish, especially crustacea and mollusks. Interestingly, tropomyosin has also been identified as an important allergen in other invertebrates including dust mites and cockroaches, and it has been proposed by some to be an invertebrate pan allergen. Different regions of shrimp tropomyosin bind IgE; 5 major IgE-binding regions have been identified in shrimp tropomyosin containing 8 epitopes. Mutations of these shrimp allergenic epitopes can reduce seafood allergenicity; methods utilizing such mutations will provide safer vaccines for more effective treatment of seafood-allergic patients, and in the future less-allergenic seafood products for consumption.

Serum insulin level, disease stage, prostate specific antigen (PSA) and Gleason score in prostate cancer.

In the present study, we assessed the relationship of serum insulin levels and three surrogate markers of recurrence, T stage, PSA, and Gleason score, in men with localized prostate cancer. Participants in our study were found through urology and radiation oncology clinics, and all eligible patients were asked to take part. All patients were asymptomatic and had been initially diagnosed on the basis of rising PSA or abnormal physical examination. Histological confirmation of diagnosis was obtained for all subjects. Serum insulin levels were determined by chemoluminescent assay with a standard, commercially available instrument. Patients were divided into three previously defined risk groups: Low risk: PSA < or =10, stage < or =T2a, or Gleason grade < or =6. Medium risk: 10 7, tumour in seminal vesicle biopsy, PSA >15 or stage T2c or T3. One hundred and sixty-three men with prostate cancer were studied. There was a significant increase in serum insulin with risk group (P=0.003, one way anova). Tukey's multiple range test showed that the insulin levels of high risk patients were significantly higher than the insulin levels of medium and low risk patients (P=0.05) but the insulin levels of medium and low risk patients were not significantly different from one another. Multivariate linear regression, with insulin as the dependent variable, Gleason score, PSA, and T stage (T1, T2, T3) as the independent variables, was significant overall (P<0.001, r(2)=0.120). Increased T stage was independently correlated with increased serum insulin levels (P<0.001). Gleason score was negatively, insignificantly correlated with serum insulin level (P=0.059). The positive correlation of PSA and insulin level was not significant (P=0.097). To assure normal distribution of insulin and PSA values, the regression was repeated with log (insulin) as the dependent variable, log (PSA), T stage (T1, T2, T3), and Gleason score as independent variables. The regression was significant overall (P=0.002, r(2) =0.095). Increased T stage was independently correlated with increased log (insulin level) (P=0.026). Gleason score was negatively, insignificantly correlated with log (insulin) level (P=0.728). The positive correlation of log (PSA) and log (insulin) levels was significant (P=0.010). The relationship between increased insulin level and advanced tumour stage in prostate cancer we describe here is biologically quite plausible, since insulin is a growth factor. Further studies may document whether serum insulin levels might be a useful biomarker of prostate cancer stage.

Identification of continuous, allergenic regions of the major shrimp allergen Pen a 1 (tropomyosin).

BACKGROUND: Crustaceans and mollusks are a frequent cause of allergic reactions. The only major allergen identified in shrimp is the muscle protein tropomyosin; at least 80% of shrimp-allergic subjects react to tropomyosin. Furthermore, tropomyosin is an important allergen in other crustaceans such as lobsters, crabs and mollusks, as well as other arthropods such as house dust mites and cockroaches, and has been implied as the cause of clinical cross-sensitivity among invertebrates. In contrast, vertebrate tropomyosins are considered non-allergenic. OBJECTIVE: The basis of the allergenicity of proteins has not yet been resolved. Thus, tropomyosin molecules provide an excellent opportunity to study the relationship between protein structure and allergenicity. The aim of the current study was to identify the IgE-binding regions of Pen a 1 and compare these regions with homologous sequences in other allergenic and non-allergenic tropomyosins. METHODS: Forty-six overlapping peptides (length: 15 amino acids; offset: 6 amino acids) spanning the entire Pen a 1 molecule were synthesized and tested for IgE antibody reactivity with sera from 18 shrimp-allergic subjects to identify the IgE-binding regions of shrimp tropomyosin. RESULTS: Based on the frequency and intensity of the IgE reactivities, five major IgE-binding regions were identified. All five major IgE-binding regions were 15-38 amino acids long. The major IgE-binding regions identified were: region 1: Pen a 1 (43-57); region 2: Pen a 1 (85-105); region 3: Pen a 1 (133-148); region 4: Pen a 1 (187-202), and region 5: Pen a 1 (247-284). In addition, 22 peptides were categorized as minor IgE-binding regions, and 12 peptides did not bind any IgE antibodies. No substantial differences in amino acid group composition in the five IgE-binding regions compared to the whole molecule were detected. Sequence identities and similarities of the Pen a 1 IgE-binding regions with homologous regions of allergenic arthropod tropomyosins were as high as 100%, whereas identities and similarities with homologous vertebrate sequences ranged from 36 to 76% and 53 to 85%, respectively. CONCLUSION: Five major IgE-binding regions of the allergenic shrimp tropomyosin, Pen a 1, were identified which are positioned at regular intervals of approximately 42 amino acids (7 heptads), suggesting a relationship with the repetitive coiled-coil structure of the tropomyosin molecule. The high degree of similarity between Pen a 1 IgE-binding regions and homologous sequences in invertebrate tropomyosins and the lower percentage of similarity with homologous regions of vertebrate tropomyosins supports a structural basis for cross-reactivity of allergenic tropomyosins.