伤寒LPS-PHA快速诊断试剂盒的评价

本文对伤寒LPS-PHA诊断试剂盒进行现场应用评价.来自各医院和不同地区977份不同类型的血清标本作LPS-PHA检测的同时与Widal试验作比较,结果伤寒沙门氏菌血培养阳性46份血清标本前者检出率(86.96％)显著高于后者(67.39％),两者在统计学上有显著差异(X~2=4.999,P<0.025),95份Widal试验阳性血清LPS-PHA阳性率为97.89％,漏检2份系乙型副伤寒抗体阳性者.607份对照血清标本LPS-PHA的假阳性率1.48％,低于Widal试验(4.84％).伤寒LPS-PHA的敏感度和特异度(94.33％和98.52％)高于Widal试验(89. 36％和95.72％).其他5项评价指标(阳性和阴性预示值,粗一致性,调整一致性和约登指数)亦都较Widal试验好.因此,LPS-PHA诊断试剂盒能满足目前临床上早期快速诊断伤寒的要求.但不适用于追溯诊断和血清流行病学调查.     

Autoimmune thyroid disorders—An update

Background: Autoimmune thyroid disease (AITD), a common organ specific autoimmune disorder is seen mostly in women between 30–50 yrs of age. Thyroid autoimmunity can cause several forms of thyroiditis ranging from hypothyroidism (Hashimoto’s thyroiditis) to hyperthyroidism (Graves’Disease). Prevalence rate of autoimmune mediated hypothyroidism is about 0.8 per 100 and 95% among them are women. Graves’ disease is about one tenth as common as hypothyroidism and tends to occur more in younger individuals. Both these disorders share many immunologic features and the disease may progress from one state to other as the autoimmune process changes. Genetic, environmental and endogenous factors are responsible for initiation of thyroid autoimmunity. At present the only confirmed genetic factor lies in HLA complex (HLA DR-3) and the T cell regulatory gene (CTLA 4). A number of environmental factors like viral infection, smoking, stress & iodine intake are associated with the disease progression. The development of antibodies to thyroid peroxidase (TPO) thyroglobulin (TG) and Thyroid stimulating hormone receptor (TSH R) is the main hallmark of AITD. Circulating T Lymphocytes are increased in AITD and thyroid gland is infiltrated with CD4+ and CD8+ T Cells. Wide varieties of cytokines are produced by infiltrated immune cells, which mediate cytotoxicity leading to thyroid cell destruction. Circulating antibodies to TPO and TG are measured by immunofluorescense, hemagglutination, ELISA & RIA. TSHR antibodies of Graves’ disease can be measured in bioassays or indirectly in assays that detect antibody binding to the receptor.     

The influence of tobacco smoking on humoral immune response in insulin dependent diabetic pregnancy

Tobacco smoking products have a heavy impact on the public health of developed as well as non-developed countries by being a main etiologic factor for the induction of cardiovascular diseases and tobacco-related cancer. The purpose of this study was to determine the influence of tobacco smoking on the measurement of the humoral immune response in Egyptian pregnant women with type 1 diabetes. Concentrations of serum immunoglobulin A, G and M in 35 smoking, 35 non-smoking pregnant women with type 1 diabetes and 35 matched normal women were measured by ELISA. Women were matched by age and working life with controls. Measurements suggested that diabetic smokers had decreased levels of IgG and IgM in their sera. It was found that normal individuals had mean IgA, IgG and IgM levels of 2.80 mg/ml, 9.33 mg/ml and 1.66 mg/ml, respectively while non-smoker women suffering from type 1 diabetes had mean levels of 3.47 mg/ml, 10.97 mg/ml and 2.05 mg/ml (p<0.0004,p<0.0001 andp<0.0002). However, the mean level of IgA, IgG and IgM in diabetic smoker sera was determined to be 3.33 mg/ml, 8.07 mg/ml and 1.31 mg/ml, respectively (p<0.003,p<0.0001 andp<0.0001). The obtained results suggest that toxic smoke components were immuno-suppressant and may well play a part in the complex immuno-pathogenesis interaction. The increased risk of smoking in insulin dependent diabetic pregnant women during pregnancy is a further reason to encourage pregnant women to quit tobacco smoking.     

Dyslipidemia in pregnancy may contribute to increased risk of neural tube defects -a pilot study in north Indian population

Neural tube defects are congenital structural abnormalities of the brain and vertebral column resulting from improper or non-timely closure of the neural tube. Prevalence of neural tube defects is reported to be higher among women with diabetes mellitus and obesity. This study was designed to investigate the relation between the presence of dyslipidemia in antenatal patients and the risk of fetal neural tube defects. The present study was an observational, cross-sectional study involving 129 pregnant women in 16 to 18 weeks gestation period. Of these, 80 women had normal pregnancies and 49 were clinically high-risk cases for neural tube defects. Fasting blood samples were analyzed for blood sugar and lipid profile by enzymatic assay and alpha-fetoprotein levels using Enzyme Immunoassay. Alpha-fetoprotein (AFP) values were converted to Multiples of Median (MoM) appropriate for the gestational age. Based on AFP values, women were labeled as screen negative (AFP <2 MoM, n= 102) and screen positive (AFP > 2 MoM, n =27). Screen positive women were further evaluated by ultrasound and 21 women were found to carry a neural tube defects positive pregnancy. Statistical analysis was done on SPSS software. Body weight of the women showed a significant positive correlation with serum triglycerides, plasma sugar and AFP MoM values. A significant difference was observed in serum cholesterol levels (p= 0.038), triglycerides (p=0.001) and plasma sugar levels (p=0.002) between normal women and those with neural tube defects positive pregnancy. The Odds ratio for neural tube defects risk in dyslipidemic cases was 24.23 (CI 4.73 − 148.60) with a relative risk of 12.12. Dyslipidemia especially hypertriglyceridemia was found to be significantly associated with fetal neural tube defects.     

HLA Gene in 5 Cases of Liver Injury Related to Baikal Skullcap

HLA association with drug-induced liver injury has recently been pointed out about multiple medicines. The aim of this study was to evaluate relationship between HLA gene and liver injury related to Baikal skullcap-containing Kampo medicines (BSCK). We previously examined HLA genes in 3 cases of BSCK-induced liver injury. Recently we could encounter 2 cases diagnosed as “definitely-related case” of BSCK-induced liver injury. HLA genes of the 2 cases were analyzed by Sequencing Based Typing method with Next Generation Sequencer at HLA Laboratory in Kyoto. HLA-DPA1*02:02:02 and DPB1*05:01:01 were observed in the 2 cases: concordance was not observed in HLA-A, B, C, DRB1, DRB4, DQA1, or DQB1. The previous 3 cases of BSCK-induced liver injury had the same allele type to the 2 cases only in HLA-DPA1. Putting all these together, HLA-DPA1*02:02:02 was observed in common among 5 cases of BSCK-induced liver injury. HLA-DPA1*02:02:02 is possibly associated with BSCK-induced liver injury.     

Effect of soy proteins Vs soy isoflavones on lipid profile in postmenopausal women

Soy isoflavones and soy proteins are being considered as possible alternatives to postmenopausal hormone replacement therapy. This study was undertaken to evaluate effects of these two preparations on symptoms and lipid profile in postmenopausal women. The study was done in 75 postmenopausal women with FSH levels = 30 mIU/ml. These women were randomly divided into 3 groups (n=25). Study group I was given soy proteins 30gm/day containing 60 mg soy isoflavones. Study group II was given soy isoflavones (60 mg/day). The control group was given casein protein 30 gm/day. The menopausal symptoms were assessed by Kupperman Index. Fasting blood samples were analyzed for serum lipid profile, apolipoprotein A1 and B, Leutenizing Hormone (LH) and Follicle-Stimulating Hormone (FSH) at the beginning of therapy, 4 and 12 weeks after initiation of therapy. A highly significant improvement in postmenopausal symptoms was observed in both the study groups. A highly significant improvement was seen in serum lipid profile and Apolipoprotein A1 and B in women taking soy proteins whereas women taking soy isoflavones demonstrated significant improvement in serum triglycerides only. Both soy proteins and soy isoflavones are helpful in alleviating postmenopausal symptoms but soy proteins offer a greater health advantage due to their beneficial effect on serum lipid profile.     

Paraoxonase 1 activity and phenotype distribution in premenopausal and postmenopausal women

Introduction

Postmenopausal women have higher risk of cardiovascular disease. One of the contributing factors could be reduced activity of anti-atherogenic enzyme paraoxonase 1 (PON1). The aim of this study was to examine differences in the lipid status, paraoxonase and arylesterase PON1 activities and PON1 phenotype in women with regular menstrual cycle and in postmenopausal women.

Materials and methods:

The study included 51 women in reproductive age (25 in follicular and 26 in luteal phase of the menstrual cycle) and 23 women in postmenopause. Lipid parameters in sera were determined using original reagents and according to manufacturer protocol. PON1 activity in serum was assessed by spectrophotometric method with substrates: paraoxon and phenylacetate. PON1 phenotype was determined by double substrate method.

Results:

Compared to the women in follicular and luteal phase, postmenopausal women have significantly higher concentration of triglyceride [0.9 (0.7–1.3), 0.7 (0.6–1.0) vs. 1.5 (0.9–1.7) mmol/L; P = 0.002], cholesterol [5.10 (4.78–6.10), 5.05 (4.70–5.40) vs. 6.30 (5.73–7.23) mmol/L; P < 0.001], LDL [3.00 (2.56–3.63), 3.00 (2.70–3.70) vs. 3.90 (3.23–4.50) mmol/L; P < 0.001], and apolipoprotein B [0.88 (0.75–1.00), 0.79 (0.68–1.00) vs. 1.07 (0.90–1.24) mmol/L; P = 0.002]. PON1 basal [104 (66–260), 106 (63–250) vs. 93 (71–165) U/L; P = 0.847] and salt-stimulated paraoxonase activity [210 (131–462), 211 (120–442) vs. 180 (139–296) U/L; P = 0.857] as well as arylesterase activity [74 (63–82), 70 (54–91) vs. 70 (60–81) kU/L; P = 0.906] and PON1 phenotype (P = 0.810) were not different in the study groups.

Conclusion:

There are no differences in PON1 activity and PON1 phenotype between women with regular menstrual cycle and postmenopausal women.     

Evaluation of A60 antibodies in pulmonary and neurotuberculosis

Humoral immune response against PPD derived A60 antigen was evaluated by quantification of serum A60 antibodies in thrity healthy adults not exposed to tuberculosis (Group 1), in twenty seven healthy adults exposed to tuberculosis patients i.e. staff working in wards of tuberculosis hospital for one to thirty years (group 2), in twenty five pulmonary tuberculosis patients admitted to the Institute for Chest Diseases, Hyderabad (Group 3) and in sixty neurotuberculosis patients admitted to Neurosurgery department of our institute (Group 4). Highly significant elevation of A60 antibodies was observed in pulmonary tuberculosis patients (p<0.01) compared to healthy adult groups. A significant elevation in serum was also observed in case of neurotuberculosis group compared to both healthy groups (p<0.01). A test on A60 antibodies in serum gavv a sensitivity of 100%, specificity of 96.6%, positive predictive value of 81% and negative predictive value of 100% for pulmonary tuberculosis, whereas a sensitivity of 58%, positive predictive value of 79% and negative predictive value of 75% were noted for neurotuberculosis patients. Results of A60 antibodies in ten cerebrospinal fluids (CSF) obtained from non tuberculosis patients and thirty two CSF from patients of neurotuberculosis did not show significant elevation of antibodies. However the ninetyfive percentile value of CSF A60 antibodies was higher in neurotuberculosis (7.4 U/ml) group compared to nontuberculous group (3.8 U/ml) and the test showed a good positive predictive value (83%), very low negative predictive value (25%) and low sensitivity (63%). Serum A60 antibody assay appears to be a good serological marker available today for pulmonary tuberculosis and a supportive marker for neurotuberculosis.     

Use of antigen prepared from in vitro culture ofPlasmodium falciparum for the detection of antibodies in sera from patients exposed toPlasmodium vivax

The blood stage parasites fromin vitro cultures ofPlasmodium falciparum were employed for the detection of malaria antibodies in patient sera havingP. vivax infections employing indirect fluorescent antibody (IFA) test. The antibody titres obtained for theP. vivax infected sera ranged from 1∶128 to 1∶1024. Control slides smeared with non-immune sera did not show any fluorescence. On the basis of the results of the present study it maybe concluded that antigen prepared fromin vitro cultures ofP. falciparum cross-reacts withP. vivax antibodies.     

Detection of tubercular antibody and antigen in sera of bone and joint tuberculosis

Trichloroacetic acid (TCA) solubilized and DEAE fractionatedMycobacterium tuberculosis H₃₇Ra excretory-secretory (ES) antigen viz., Mtb EST DE1 and affinity purified goat antibodies to the TCA solubilized ES antigen (Mtb EST) were explored in detecting tubercular antibody and antigen respectively in sera of bone and joint tuberculosis by indirect and sandwich ELISA. Out of total 36 bone & joint tuberculosis cases, tubercular antibody was detected by indirect ELISA in 30 patients (sensitivity 83%), while circulating tubercular antigen was detected by sandwich ELISA in 27 patients (sensitivity 75%). Out of 34 non tubercular disease control cases, 10 patients showed positive reaction for antibody while only 4 patients showed positive reaction for antigen. In another group of 34 healthy subjects who were screened, 4 individuals showed positive reaction for tubercular antibody and 2 cases for antigen. This study shows that antigen detection assay using affinity purified anti Mtb EST antigen antibody is superior with overall specificity of 91% as compared to antibody detection assay with 75% specificity in bone & joint tuberculosis.     

Presynaptic Membrane Receptor in Human Brain

Myasthenia gravis (MG) is an autoimmune disease that results from antibody mediated damage of Acetylcholine receptor (AChR) at the neuromuscular junction. The autoimmune character of MG and pathogenic role of AChR antibodies have been established by several workers i.e., the demonstration of anti-AChR antibodies in about 90 % of MG patients. It has been demonstrated that patients with MG also have antibodies against a second protein named presynaptic membrane receptor (PsmR), which is identified by utilizing β-Bgtx, a ligand which binds to PsmR. Using β-Bgtx Sepharose 4B affinity matrix, the PsmR was purified from different regions of human cadaver brain by affinity chromatography. Purified receptor was characterized both by biochemical and immunological procedures. PsmR purified from different regions of the brain shows a specific activity of 0.37 ± 0.01, 0.39 ± 0.02 and 0.43 ± 0.005 nM/ μg of protein in Parietal lobe, Occipital lobe and Frontal lobe respectively. The affinity purified PsmR from the brain of 87 and 68 kd (parietal lobe, occipital lobe and frontal lobe) shows immunoreactivity with myasthenic sera. These findings suggest that PsmR from brain is another antigen against which autoantibodies are developed in Myasthenia gravis patients. Upon treatment with various enzymes we concluded that PsmR from brain is a glycoprotein in which the immunoreactivity resides in the carbohydrate as well as the peptide epitopes. In conclusion the PsmR is another antigen against which autoantibodies are formed in different regions of brain. These can be used as a diagnostic tool for detecting antibodies in the sera or cerebrospinal fluid of MG patients.     

Evaluation of serum adenosine deaminase and 5′-nucleotidase activities as probable markers in ovarian cancer

Adenosine deaminase (ADA) and 5′-nucleotidase (5′-NT) activities were measured in sera of patients with ovarian cancer and patients with benign ovarian tumour. The results were compared with that of a control group consisting of healthy women. ADA levels were significantly increased (P<0.001) in the ovarian cancer group (n=50) but not in the benign group (n=28) when compared to the controls (n=20). The results indicate that ADA and 5′-NT levels may help to differentiate malignant conditions from benign tumours of the ovary in addition to the existing tests such as serum CA-125 levels and histopathological study.     

Diagnosis of gastrointestinal tuberculosis: Using cytomorphological,microbiological, immunological and molecular techniques — A study from Central India

The present study included three groups: (A) age and gender matched control (n=24) with no previous signs of M. tuberculosis complex (MTBC) infection, (B) patients (n=28) diagnosed with gastro-intestinal TB (GITB), (C) patients (n=50) with clinical and histo-pathological signs of GITB, but were culture and AFB negative. Real time assay performed using fluorescence resonance energy transfer hybridization probes showed a positivity index of 36 % in group C, i.e. 18 were found reactive from the total 50 cases studied. In addition, immune characterization of these 18 cases showed depleted CD₄⁺ count and increased levels of IFN-γ and TNF-α cytokines. No positive case was found in group A, while in group B, out of total 28 cases studied 27 were found positive. A combinatorial diagnostic approach for rapid detection and characterization of GITB might provide specific therapeutic strategies for prevention and treatment of the infection in future.     

Effects of short-term hormone replacement on atherogenic indices in Indian postmenopausal women

Indian ethnicity by itself is a strong risk factor for development of CAD in Indian postmenopausal women due to lower HDL levels as compared to Whites and women of oriental origin. We evaluated and compared the short-term effects of menopause, estrogen replacement therapy and combined estrogen and progestin replacement therapy on various atherogenic indices. 40 postmenopausal women, both surgical and natural (20 each) were selected. 10 surgical postmenopausal women were given 0.625 mg conjugated estrogens daily for 6 months and 10 natural postmenopausal women were given 0.625 mg conjugated estrogen with 2.5 mg medroxyprogesterone acetate daily. 20 women were included in the control group and given placebo. Fasting venous blood samples were analyzed for extended lipid profile and calculated atherogenic indices before starting the therapy and after 1,3 and 6 months. LDL and Apolipoprotein B increased (p<0.05) and those of Apolipoprotein A1 and HDL decreased in the control groups. In both the study groups levels of serum cholesterol and LDL decreased (p<0.05) and those of HDL and Apolipoprotein A1 increased (p<0.01). LDL/HDL, Apo B/ApoA1, Total Cholesterol/HDL decreased significantly (p<0.05) in both the study groups compared to the control groups. The effect of estrogen alone was more significant as compared to combination therapy. Log Triglycerides (TG)/HDL ratio showed a decrease in women on estrogen alone but the difference was not significant. Our study confirms that short term HRT has a favorable effect on atherogenic indices in Indian postmenopausal women.     

Elisa based diagnostic test for Aspergillosis

Detection of lgE and lgG antibodies in Aspergillosis is of diagnostic significance. The serological methods, such as agglutination, gel diffusion and counter immuno electrophoresis that are commonly used in the laboratories for diagnosis of Aspergillus induced infections, are less sensitive and high cross reactivity is often encountered. We carried out work on characterization and identification of diagnostically relevant antigens ofA. fumigatus. Well characterized antigens were used to develop an ELISA with 92% sensitivity and 89% specificity for detection of specific lgE and lgG in the sera of patients of allergic bronchopulmonary aspergillosis (ABPA), aspergilloma and invasive aspergillosis. Subsequently, a sample kit having “ready to use type” of dry reagents (powder/tableted buffers and lyophilized antigen, conjugate and reference sera) was formulated. The kit was validated with sera from patients of ABPA, related allergic disorders, tuberculosis, post-Kochs cases and thalassemic children receiving repeated blood transfusions. The performance of the kit was found to be satisfactory with coded sera.     

Urinary enzymes in nephrotic syndrome

Tubular damage is a complication associated with nephrotic syndrome and increased levels of urinary enzymes are of significant value in detection of the same. The aim of our study was to evaluate the use of urinary lysozyme and trehalase as markers of tubular dysfunction in nephrotic syndrome. This study assessed 35 nephrotic syndrome patients and 30 healthy controls matched for age and sex. Urine samples were examined at pretreatment and post treatment (8 weeks) stages for proteinuria, lysozyme and trehalase. At pretreatmant stage there was significant increase in urinary lysozyme and trehalase as compared to controls (p<0.001). A good correlation was observed between degree of proteinuria and urinary lysozyme (p<0.001;r=0.80) and trehalase (p<0.001; r=0.74). At the end of 8 weeks of treatment, the patients showed significant decrease in their urinary lysozyme and trehalase activity (p<0.001) but no correlation with degree of proteinuria was observed. Our results indicate that enzymes like lysozyme and trehalase can be used as markers of tubular dysfunction.     

Antinuclear Antibody Profiling in Patients of a Tertiary Care Centre in Central India

The incidence of autoimmune disorders that includes the connective tissue diseases has seen a rise in India in recent times. Antinuclear antibodies, the telltale sign of systemic autoimmune response, thus can be used as a screening tool and also to support the diagnosis of systemic autoimmune disease. The present retrospective cross- sectional analysis aimed to study the antinuclear antibodies profile (patterns and specific antibody reactivity) amongst suspected cases of auto-immune disorders at a tertiary care teaching hospital. The study retrieved and reviewed reports of 644 patients sent for ANA testing by indirect immunofluorescence assay over a period of 1 year by different specialty departments. Positive samples were further processed for anti-ds-DNA antibody and antibodies to extractable nuclear antigen. Data collected was statistically analysed. ANA pattern positivity was observed in 31% of cases and a positive antibody reactivity was seen in 66% of them. Female predominance (82%) was noted in both pattern positivity and antibody reactivity. High levels of pattern positivity and antibody reactivity was found in the young adults (45.9%). Amongst the ANA patterns, the nuclear homogenous pattern was found the commonest. The common antibodies associated with this pattern were anti-dsDNA and U1 Sm/RNP antibodies. A stronger fluorescence intensity on initial screening showed a higher confirmation rate for specific antibodies on immunoassay. High occurrence of positive ANA patterns in autoimmune disorders suggests its utilization as a screening tool for them and would also play an adjuvant to the diagnosis. Early knowledge about future autoimmunity will earn better prognostic achievements through better treatment interventions.     

Association of Obesity with Hormonal Imbalance in Infertility: A Cross-Sectional Study in North Indian Women

Hormones play an important role in the development and regulation of reproductive function and the menstrual cycle of women. Extremes of body weight tend to affect the homeostasis of the hypothalamo–pituitary–gonadal axis. This cross-sectional study was carried out in 113 women (57 with primary infertility and 56 with secondary infertility) in the age group 20–35 years, presenting for hormonal evaluation of infertility in a tertiary care hospital. After preliminary clinical evaluation, anthropometric indices (height, weight, BMI, waist circumference and waist hip ratio) were measured in all subjects. Fasting blood sample drawn on second/third day of menstrual cycle was analysed for serum luteinizing hormone, follicle stimulating hormone (FSH), prolactin and thyroid stimulating hormone (TSH). Serum FSH levels showed a significant positive correlation with indicators of central obesity (waist circumference and waist hip ratio in both the study groups). In primary infertility, significant positive correlation was also observed between serum FSH levels and other markers of obesity like body weight, hip circumference and BMI. In secondary infertility, serum prolactin and serum TSH levels demonstrated a significant positive correlation with body weight and BMI. Obesity is associated with hormonal derangements which are responsible for infertility. In overweight women with infertility, weight loss should be considered as a first line treatment.     

Estimation of Upper Limit of Normal for Serum Alanine Transaminase in Healthy South Indian Population

Alanine aminotransferase (ALT) is the most common and cost effective screening test for asymptomatic liver disease. There is paucity of data on normal ALT among healthy individuals in India. An observational cross sectional study was conducted from January to July 2013 to estimate the upper limit of normal for ALT in healthy south Indian population. Adults undergoing voluntary or pre-employment health screening were included. Those with current and past alcoholism or smoking, acute illness or hospitalization during preceding 12 months, non-steroidal anti-inflammatory or over the counter medication use within a month, current or past intake of herbal medications, any chronic medical illness, abnormal body mass index (BMI), fatty liver in ultrasound, abnormality in haemoglobin, platelet count, blood sugar, creatinine, lipid profile and thyroid function test and positive serology (Hepatitis B, C or HIV) were excluded. A total of 2600 subjects were screened. 344 were included for analysis. Mean age was 35 years in men and 34.83 years in women, with a mean BMI of 22.2 kg/m² in men and 21.8 kg/m² in women. The mean ALT in men and women were 21.87 ± 2.9 (97.5th percentile 28 U/L) and 19.35 ± 3.3 (97.5th percentile 24 U/L) respectively. In conclusion, mean and upper limit of ALT (97.5th percentile) in south Indian men was 21.87 and 28 IU/L and women were 19.35 and 24 IU/L respectively. There is a need to re-consider ALT levels in our population for better detection of individuals at risk for liver disease.     

Antibodies against free radical modified DNA in cancer patients

In this study we have modified DNA by exposing it to ultraviolet light in the presence of hydrogen peroxide. The modified DNA was probed for binding to the antibodies present in the sera of patients suffering from various types of cancer. Higher recognition of modified DNA, as compared to native DNA, by antibodies from cancer patients has got far reaching significance.