Gender differences in homocysteine concentrations, a population-based cross-sectional study.

BACKGROUND AND AIMS: High concentrations of homocysteine are considered a risk factor for atherosclerosis and coronary artery disease. The aim of this study was to assess whether or not there are gender differences in the plasma concentrations of homocysteine. METHODS AND RESULTS: Data were collected from medical records of individuals examined at a screening center in Israel between the years 2000-2014. Cross sectional analysis was carried out on 9237 men and 4353 women. Mean (SD) age of the study sample was 48.4 (9.7) and 47.7 (9.7) years for men and women respectively. Average homocysteine concentrations were 12.6 (5.9) and 9.6 (3.2) µmol/L in men and women respectively (p < 0.001). Prevalence of homocysteine concentrations above 15 µmol/L was found to be significantly higher in men than in women; 15.5% vs 3.9% respectively (p < 0.001). Low concentrations of vitamin (B12 < 200 pmol/L) and low concentrations of folate (<12 nmol/L) were found to be significantly higher in men than in women 20.4% vs. 16.0% and 18.5% vs. 10.8% respectively. Compared to women, men had a significantly higher odds ratio (95% CI) of having homocysteine concentrations above 15 µmol/L: non adjusted model, 4.47 (3.80-5.26); adjusted model for age, smoking status, body mass index, diabetes mellitus, kidney function and low serum concentrations of vitamin B12 and folate, 3.44 (2.89-4.09). CONCLUSION: Plasma homocysteine concentrations are higher in men than in women. This may be a contributing factor to gender differences for developing atherosclerosis and coronary artery disease.

Cross-reactive epitopes on beta2-glycoprotein-I and Saccharomyces cerevisiae in patients with the antiphospholipid syndrome.

Anti-Saccharomyces cerevisiae antibodies (ASCA), directed against the phosphopeptidomannan (PPM) part of the cell wall of the yeast, have been identified as an important and specific serological marker for Crohn's disease. We evaluated the prevalence and properties of ASCA in APS patients. Thirty-one out of 155 APS patients tested positive for ASCA (20.0%), compared to 5.0% in healthy controls (P < 0.05). The presence of ASCA was not associated with any specific manifestation of APS. The ASCA found to be the population of anti-beta2GPI antibodies (Abs). Affinity purified anti-beta2GPI from ASCA-positive sera on a beta2GPI column, bound specifically the PPM, as shown by direct binding and competition assays (95-98%). The PPM inhibited differentially the anti-beta2GPI binding to beta2GPI. Since the anti-beta2GPI anti-PPM could bind only native form of beta2GPI and not the recombinant form, we assume that these specific anti-beta2GPI subpopulations of Abs are directed to the glycosylated site of the molecule. In conclusion, a subpopulation of anti-beta2GPI is specific to the glycosylated site of the beta2GPI molecule that cross-reacts with PPM.

Anti-Saccharomyces cerevisiae antibodies in Behçet's disease--a familial study.

OBJECTIVE: To evaluate the rate and clinical correlations of antibodies against saccharomyces cerevisiae (ASCA) among healthy family members of patients with Behçet's disease (BD). METHODS: Twenty-one BD patients and 52 healthy family members (HFM) were studied. Data from medical files and from patients' interviews was collected, regarding the entire spectrum of disease manifestations. Each family member was personally interviewed and a questionnaire composed of BD symptoms and their temporal relation was compiled. IgA- and IgG-ASCA levels, determined by ELISA, were studied in all BD patients and their family members, the results were compared to a group of 23 healthy controls (HC). RESULTS: Eight (38.1%) BD patients were ASCA positive, compared to five among HFM (9.6%) and none among healthy unrelated controls (p=0.001). Mean IgG and IgA-ASCA levels were significantly higher in BD patients compared with HFM and HC groups (p = 0.002 and p = 0.03, respectively). No correlation was found between positive ASCA tests and any of BD-related manifestations. Mean IgG-ASCA levels were significantly lower in HFM compared to BD patients (p = 0.03), yet IgA-ASCA levels were similar in HFM and BD. Mean IgG and IgA-ASCA levels were higher in HFM compared with healthy unrelated controls (p=0.09 and p=0.03). No difference was found in ASCA rates between relatives of BD patients who had positive or negative ASCA tests, or between spouses of BD patients and genetically related relatives. In HFM with recurrent oral ulcers there was a positive correlation between titers of IgA-ASCA and the yearly number of oral ulcers episodes (p = 0.01), and mean ulcers healing time (p = 0.01). IgG-ASCA titers correlated with yearly number of aphtae episodes (p = 0.03). CONCLUSION: The results of this study confirm our previous observation on a high prevalence of ASCA in BD. ASCA levels are also increased in healthy family members of BD patients, and are probably influenced by genetic as well as environmental factors. ASCA in HFM were significantly associated with a more severe oral ulcer disease. The role of ASCA as a marker for predisposition to develop future BD remains to be evaluated.

Immunomodulation of experimental autoimmune diseases via oral tolerance.

The concept of oral tolerance refers to a form of peripheral tolerance in which mature lymphocytes in the peripheral lymphoid tissues are rendered nonfunctional or hyporesponsive by prior oral administration of an antigen. The primary mechanisms mediating oral tolerance include deletion, anergy of antigen-specific T cells and active cellular suppression, the primary determining factor being the dose of fed antigen. Low doses favor active suppression, whereas high doses favor deletion and anergy. Active cellular suppression is mediated by the induction of regulatory T cells in the gut-associated lymphoid tissue, which migrate to the systemic immune system. One of the primary mechanisms of active cellular suppression is via secretion of suppressive cytokines such as TGF-beta, IL-4, and IL-10 following antigen-specific triggering. TGF-beta is produced both by CD4+ and CD8+ GALT-derived T cells and is an important mediator of the active suppression component of oral tolerance. CD4+ cells that primarily produce TGF-beta appear to be a unique T-cell subset and termed Th3 cells. Oral tolerance was successfully studied in a variety of experimental models for autoimmune diseases, among them experimental autoimmune encephalomyelitis, experimental arthritis, experimental anti-phospholipid syndrome, experimental autoimmune uveoretinitis, experimental insulin dependent diabetes mellitus (IDDM), and experimental autoimmune myasthenia gravis. The results obtained in experimental animal models have led to the conduction of several clinical trials of oral tolerance in patients with multiple sclerosis, rheumatoid arthritis, uveitis, and IDDM. Conflicting results were obtained, and although some improvement has been noted in some of the patients, broad ranging clinical improvement has not yet been observed. A more accurate choice of antigens, as well as more precise dosing and timing of antigen-administration might lead to better results in the future.

Livedo reticularis is a marker for predicting multi-system thrombosis in antiphospholipid syndrome.

OBJECTIVE: Livedo reticularis (LR) is a skin vasculopathy that has been frequently described in patients with anti-phospholipid syndrome (APS) and reported to be present in association with valvular heart pathology and strokes (i.e. Sneddon's syndrome). METHODS: In a cohort of APS patients we investigated the possible association of LR with various clinical aspects of APS such as pregnancy morbidity, central nervous system (CNS) and cardiac manifestations. RESULTS: Livedo reticularis was found in 50/308 (16%) of APS patients, and there was a significant association with cerebrovascular accidents (CVA), migraines and epilepsy (p = 0.01, 0.002, and 0.02 respectively). A similar association was also detected between LR, and the presence of cardiac valve thickening and vegetations (p = 0.001). No association with venous thrombosis, recurrent fetal loss, IUGR or toxemia was found. CONCLUSION: Livedo reticularis is a frequent cutaneous manifestation in patients with APS. Its high association with cardiac and CNS thrombosis may suggest that LR-APS patients compose a subset at higher risk for thrombosis, and thus may require a closer follow-up and a more aggressive anticoagulation.

Bovine seminal plasma aSFP: localization of disulfide bridges and detection of three different isoelectric forms.

Acidic seminal fluid protein (aSFP) is a major 13 kDa protein isolated from bull seminal plasma and characterized as a new growth factor which stimulates in vitro cell division and progesterone secretion by ovarian cells. Here, we establish that the four cysteines of oxidized aSFP form two disulfide bridges between nearest-neighbour residues. This pattern is conserved in boar spermadhesins, with which aSFP shares up to 50% amino acid sequence identity, and other proteins of the recently identified CUB domain family. Using isoelectric focusing in combination with sulfhydryl group-specific blotting, the three forms of aSFP were identified as completely oxidized (pI 4.7), partly reduced (pI 4.8) and fully reduced at pI 5.1. These results indicate that native aSFP possesses two pairs of cysteine residues of different reactivity. The observation that aSFP can protect sperm from oxidative damage might be explained by its reduction/oxidation behaviour.

10th International Congress on Antiphospholipid Antibodies--summary.

The 10th International Congress on Antiphospholipid Antibodies (Sicily, Italy, September 29-October 3, 2002) (Fig. 1) provided enlightening aspects on the recent developments in antiphospholipid syndrome (APS) and antiphospholipid antibodies in more than 150 lectures and posters. Researchers from all aspects of medicine attended the meeting, implicating the systemic characteristics of APS. The important breakthroughs are summarized.

Possible second case of tricho-oculo-dermo-vertebral (Alves) syndrome.

We report on a 10-year-old girl with trichodysplasia, onychodysplasia, hyperpigmented ichthyoform lesions on her shins, mild enamel dysplasia, and hyperkeratosis involving the soles. This combination of ectodermal defects has only been described by Alves et al.

Anti-endothelial cell antibodies (AECA) in systemic sclerosis--increased sensitivity using different endothelial cell substrates and association with other autoantibodies.

OBJECTIVE: One of the main features of systemic sclerosis (SSc) is vascular damage, the mechanism of which is not understood. In the present study we examined whether screening of SSc patients for different anti-endothelial cells antibodies (AECA) of various origins increase the sensitivity of AECA detection in SSc patients. Secondary aim was an attempt to correlate AECA with other common autoantibodies. MATERIALS & METHODS: 478 SSc patients were studied for the presence AECA, anti-cardiolipin (aCL), anti-dsDNA, anti-heparin (AHA), anti-pyruvate dehydrogenase (PDH) and anti-PDC-E2 autoantibodies. AECA levels were detemined using human umbilical vein EC (HUVEC), bone marrow EC (BMEC), EC hybridoma (EA.hy 926) and Kaposi sarcoma EC (KS). RESULTS: Positive AECA were found in 49.5% of SSc patients (27.1% HUVEC; 34.3% BMEC; 26.3% EaHy 926 and 22.7% KS). The highest percent reactivity of AECA was obtained using microvascular BMEC. When combining BMEC and either other cell lines the reactivity ranged from 41.4% to 46%. A significant association between AECA on the one hand and AHA (p<0.001)) and anti-PDH (p<0.05) on the other was secn. Cross-reactivity with anti-PDC-E2 was excluded by inhibition tests, but AHA and anti-PDH may be part of the spectrum of AECA. CONCLUSIONS: Since false-negative AECA may result from lack of expression of various antigens on a specific EC, analysis of AECA in SSc patients requires using several EC types, including microvascular EC.

Peptide immunotherapy in autoimmune diseases.

Peptide-mediated immunotherapy has been studied in a number of experimental models of autoimmune diseases and has also been tested in human patients to a certain extent. Copolymer 1 is a synthetic amino acid copolymer that has been demonstrated to suppress experimental autoimmune encephalomyelitis (a model for multiple sclerosis) when administered parenterally. Some study results indicate that mucosal tolerance induced by appropriate recombinant peptide fragments of human AChR is effective in suppressing experimental autoimmune myasthenia gravis and might be considered as a therapeutic modality for patients with MG. A peptide of the heat-shock protein 60 molecule, designated peptide p277, was shown to be a target of T cells in autoimmune diabetes in NOD mice, and intraperitoneal injections of glutamic acid decarboxylase (GAD) peptide 524-543 delayed the onset of diabetes and significantly reduced its incidence. Experimental evidence has revealed that CDR-based peptides may be potential candidates for the therapy of systemic lupus erythematosus. The use of synthetic peptides that focus on neutralization of pathogenic anti-beta 2GPI antibodies represents a possible new therapeutic approach to antiphospholipid syndrome. Studies in both acute and chronic-relapsing experimental autoimmune uveoretinitis have indicated that oral administration of S-Ag, S-Ag-derived peptides, inter-photoreceptor retinoid binding protein or HLA-derived peptides before immunization can protect animals from the disease.

Pathergy reaction in Behçet's disease: lack of correlation with mucocutaneous manifestations and systemic disease expression.

OBJECTIVE: The pathergy reaction is a unique feature of Behçet's disease (BD) and, according to the International Study Group (ISG), is among the major criteria required for the diagnosis. Different positive pathergy reaction rates in BD have been reported worldwide. We evaluated the prevalence of the pathergy reaction in Israeli BD patients, and its relation to mucocutaneous and systemic manifestations of the disease. METHODS: Forty-three patients were studied, all of whom fulfilled the ISG criteria for BD. The mucocutaneous and systemic disease manifestations were analyzed with respect to the presence of the pathergy reaction, and a systemic severity score for BD was calculated according to the potential morbidity and mortality associated with various clinical features. RESULTS: Nineteen patients (44.2%) had a positive pathergy test. The pathergy-positive and pathergy-negative BD groups showed a similar male:female ratio, age at disease onset, and mean disease duration. They also exhibited similar HLA-B5 levels and a similar frequency of oral ulcerations in close family members. The mucocutaneous manifestations, systemic disease expression, and severity score were similar in patients with and without the pathergy reaction. CONCLUSION: The presence of a positive pathergy reaction, although common in Israeli BD patients, is not associated with an increased risk for specific mucocutaneous or systemic manifestations of the disease, and probably does not predict a more severe disease course.

Anti-Saccharomyces cerevisiae antibodies--a novel serologic marker for Behçet's disease.

OBJECTIVE: To evaluate the prevalence and clinical correlations of antibodies against Saccharomyces cerevisiae (ASCA) among patients with BD. METHODS: Twenty-seven BD patients were studied. Data from medical files and from patients' interviews was collected, regarding the entire spectrum of disease manifestations, and a severity score was calculated for each patient. IgA- and IgG-ASCA levels, determined by ELISA, were studied in all BD patients and in three control groups: patients with recurrent aphthous stomatitis (RAS), systemic lupus erythematosus (SLE) and healthy volunteers. RESULTS: Thirteen BD patients (48.1%) were ASCA-positive, compared to one patient in each control group (10%, p = 0.01). The mean value of IgG-ASCA in the BD patients was 20.7 +/- 12.3 units, significantly higher than in patients with RAS (10.0 +/- 5.5, p < 0.001), SLE (11.8 +/- 9.3, p < 0.03) or healthy volunteers (10.8 +/- 9.8, p < 0.02). Mean IgA-ASCA level was 16.8 +/- 8.8 units in the BD patients, significantly higher compared to healthy volunteers (11.0 +/- 5.0, p = 0.02) but similar to patients with RAS (17.0 +/- 5.3). No correlation was found between ASCA and any BD-associated clinical manifestation nor the presence of HLA-B5. No difference was found in the rate of major oral ulcers nor in the systemic disease severity score between positive- and negative-ASCA patients (27.3% vs. 30.8%, and 7.31 +/- 1.80 vs. 7.28 +/- 2.27 respectively, NS). CONCLUSION: The results of our study associate, for the first time, the presence of a distinct antibody, i.e. ASCA, with BD. ASCA were not linked to a specific clinical manifestation of the disease and probably do not pose an increased risk for a more severe disease course.

Disease patterns of patients with Behçet's disease demonstrated by factor analysis.

OBJECTIVE: To explore the main patterns of Behçet's disease (BD) expression, applying factor analysis. METHODS: Sixty-eight BD patients were studied. The following disease manifestations were used for the factor analysis: genital ulcerations, typical skin lesions (erythema nodosum, folliculitis or papulo-pustular rash), uveitis, CNS involvement, joint disease, deep vein and superficial vein thrombosis, and gastrointestinal manifestations. The results were further analyzed according to sex, HLA typing, and childhood vs. adult-onset disease. RESULTS: Five factors were derived, which accounted for 69% of the variance of the matrix. Factor 1 represented the association between folliculitis and genital ulceration. Factor 2 represented the association between papulo-pustular rash and gastrointestinal symptoms. Factor 3 represented the inverse association between superficial vein thrombosis and erythema nodosum. Factor 4 represented the correlation between deep vein thrombosis and neuro-Behçet. Factor 5 represented joint disease. No difference was found between males and females in relation to factors 1, 2 or 5, but factors 3 and 4 had higher scores in male patients (p = 0.1 and p = 0.07, respectively). Factor 3 was significantly higher in patients with HLA-B5, compared to HLA-B5-negative BD patients (p < 0.001). Factors 1 and 3 were higher in patients with adult onset of the disease (p = 0.07, and p = 0.003, respectively), while factor 2 was higher in patients with childhood-onset BD (p = 0.07). CONCLUSIONS: The application of factor analysis revealed possible associations between distinct types of skin lesions, or venous thrombosis, and other disease manifestations of Behçet's syndrome, some of which were sex, age at onset, or HLA-related.

Improved method for the simultaneous determination of whey proteins, caseins and para-kappa-casein in milk and dairy products by capillary electrophoresis.

A capillary electrophoresis method for the simultaneous determination of whey proteins, caseins and their degradation products, such as para-kappa-casein, was proposed. The effect of several parameters (pH, ionic strength and concentration of urea in the electrophoresis buffer and applied voltage) on the analysis time and on the separation efficiency of the major milk proteins was studied. Using a hydrophilically coated capillary, in combination with electrophoresis buffer 0.48 M citric acid-13.6 mM citrate-4.8 M urea at pH 2.3, and a separation voltage of 25 kV, a complete separation of beta-lactoglobulin and para-kappa-casein was achieved, permitting the quantification of both components.

Sinking heart: a Punjabi communication of distress.

Within transcultural psychiatry there is a continuing debate between universalist and relativist positions. This debate focuses on the translation of Western psychiatric categories to non-Western languages and cultural contexts, and on the cross-cultural applicability of a Western model of human nature. This debate is of concern to medical practitioners and other primary carers who work with ethnic minority patients. The paper describes a syndrome of heart distress referred to as "sinking heart' by Punjabis living in Bedford, and it discusses how far this condition correlates with Western psychiatric categories. "Sinking heart' is an illness in which physical sensations in the heart or in the chest are experienced and these symptoms are thought to be caused by excessive heat, exhaustion, worry and/or social failure. The Punjabi model of "sinking heart' offers a culture-bound explanation of somatic symptoms. It is based on culturally specific ideas about the person, the self and the heart and on the assumption that physical, emotional and social symptoms of pathology accompany each other. This model is compared with Western notions of depression and with medical models of heart distress, type A behaviour pattern and stress. The paper concludes that the Punjabi model of sinking heart does not exactly correspond to any of these. The sinking heart model bears closest resemblance to a Western model of stress. The similarity between these two models is in the form rather than in the content.

MRI meningeal enhancement with intracranial hypotension caused by lumbar puncture.

A 12-year-old girl with recent history of pseudotumor cerebri developed severe postural headache due to intracranial hypotension after lumbar puncture. Meningeal enhancement and thickening were demonstrated on magnetic resonance imaging. Repeated imaging 3 months later disclosed no abnormalities. This is the first report of a child with evident meningeal enhancement on magnetic resonance imaging related to lumbar puncture.

A rapid microbore HPLC method for determination of primary structure of beta-lactoglobulin genetic variants.

A rapid method for determination of the primary structures for beta-lactoglobulin (beta-LG) genetic variants is described. This included rapid microbore HPLC, amino acid analyses, and wherever necessary, direct peptide sequencing. Two novel variants of beta-LG have been identified, bovine beta-LG W and ovine beta-LG C. The proteins were oxidized, digested with trypsin and separated using RP-HPLC. All peptides were recovered in a single run. Peptides with amino acid exchanges were identified by retention time and subjected to amino acid and sequence analyses. Ovine beta-LG C differs from the ovine beta-LG A variant by a single amino acid exchange at position 148 where Arg is replaced by Gln. Bovine beta-LG W differs from bovine beta-LG B by having Leu at position 56 instead of Ile. The method described here is reliable and can be used for mapping of 20-1000 pmol of material.

Cytokine levels in various intravenous immunoglobulin (IVIg) preparations.

The mechanisms of action of intravenous immunoglobulin (IVIg) in autoimmune diseases include modulation of cytokine levels. We examined therefore whether direct infusion of abnormally high levels of 13 different cytokines or cytokine-inhibitors within 5 different IVIg preparations have any role in modulation of their levels. None of the measured cytokines in any of the IVIg preparations tested were above the normal levels, and regarding some no traces could be detected. Hence, modulation of cytokine levels following IVIg therapy involves other mechanisms such as interference with their secretion or cytokine-specific blocking antibodies, rather than direct infusion of cytokines.

[Non-ketotic hyperglycinemia]. / Non-ketoticus hyperglycinaemia.

Three children with non-ketotic hyperglycinaemia (NKH) is reported. Two patients had typical neonatal form of NKH, one patients had atypical form of NKH. The clinical symptoms laboratory findings and therapeutical approach are discussed. One of the patients with typical neonatal form of NKH is died, neuropatological examination revealed corpus callosal agenesis and diffuse hypomyelinisation. The two children treated with N-methyl-D-aspartate-antagonist drugs reached a significantly better clinical condition. The authors reviewed the data of the literature, especially focused on the therapeutical possibilities.