Development of a capsid based competitive inhibition enzyme-linked immunosorbent assay for detection of bovine immunodeficiency virus antibodies in cattle and buffalo serum

The aim of this study was to develop a more specific and sensitive competitive inhibition ELISA (CI-ELISA) than the currently used indirect ELISA for detection of antibodies to bovine immunodeficiency virus (BIV) in cattle and buffaloes. Murine monoclonal antibodies (MAbs) were generated against a recombinant capsid (CA) protein of bovine immunodeficiency virus. Of the 13 anti-CA MAbs developed, MAb-9G10 was selected for CI-ELISA based on the maximum inhibition (98%) obtained with reference BIV antibody positive serum. Based on the distribution of percent inhibition of known negative sera (n=50), a cut-off value was set at 40% inhibition. The MAb-based CI-ELISA showed much higher agreement (concordance: 95.4%) than the indirect ELISA (concordance: 77.8%) with Western blot. Out of 672 sera of cattle and buffaloes tested by CI-ELISA from four states of India, 22% (113/516) of cattle and 19% (30/156) of buffalo were sero-positive for BIV with an overall seroprevalence of 21% (143/672) in India.     

Methylphenidate for treating tobacco dependence in non-attention deficit hyperactivity disorder smokers: A pilot randomized placebo-controlled trial

Background  

Methylphenidate blocks the re-uptake of dopamine by binding to the dopamine transporter in the presynaptic cell membrane and increases extracellular dopamine levels. Similarities in neuropsychologic effects between nicotine and methylphenidate make it an intriguing potential therapeutic option. Previous research of methylphenidate in smokers has suggested a possible beneficial effect for the relief of nicotine withdrawal symptoms, but showed no efficacy in helping smokers with attention deficit hyperactivity disorder (ADHD) to stop smoking.     

Iron food supplement

OBJECTIVE: To develop an iron rich supplement with locally available foods and to test its feasibility in school going children (7-9 Years) belonging to low income families. METHODS: From the upper primary school in Rajendranagar 7-9 year-old children were screened for hemoglobin (Hb) levels and 36 children having Hb levels below 11 g/dl were selected. Based on their Hb levels, age, and gender, 24 children were grouped as experimental and the rest as control. A supplement food (laddoo) was developed using locally available foods like jaggery, processed rice flakes, graden cress seeds and amaranth seeds (45:40:10.5). In the experimental group, children were given one laddoo per day for a period of 60 days. Effect of supplement on Hb levels, height and weight was assessed. RESULT: Anthropometric parameters showed that 97% of these children were undernourished, majority (50%) were in grade II malnutrition followed by grade (25%) and grade II (22%). A significant increase in Hb levels was observed in both the boys and girls after 30 days of supplementation only. The increase was comparatively more in the first 30 days than the second 30 days. The overall increase in Hb levels was more in 7-8 years than the 8-9 years age group. In majority of the subjects progression from one Hb level to the next higher level was observed. There was no significant improvement in their height and weight. CONCLUSION: The product developed contributed around 39-mg% iron. Thus its efficacy as an iron rich supplement in combating iron deficiency anemia is reflected in the results obtained.     

Distinctive DNA methylation patterns of cell-free plasma DNA in women with malignant ovarian tumors

Objective

Epithelial ovarian carcinoma (OvCa) is rarely detected early, and it is also difficult to determine whether an adnexal mass is benign or malignant. Previously, we noted differences in methylation patterns of cell-free plasma DNA (cfpDNA) in women without disease compared to patients with OvCa. In this work, we investigated whether methylation patterns of cfpDNA can differentiate between benign and malignant tumors.

Methods

Methylation patterns in cfpDNA were determined in three cohorts (30 samples each) using a microarray-based assay (MethDet 56). Principal component analysis, supervised clustering, linear discrimination analysis, and 25 rounds of 5-fold cross-validation were used to determine informative genes and assess the sensitivity and specificity of differentiating between OvCa vs. healthy control (HC), benign ovarian disease (mostly serous cystadenoma, BOD) vs. HC, and OvCa vs. BOD samples.

Results

Differential methylation of three promoters (RASSF1A, CALCA, and EP300) differentiated between OvCa vs. HC with a sensitivity of 90.0% and a specificity of 86.7%. Three different promoters (BRCA1, CALCA, and CDKN1C) were informative for differentiating between BOD vs. HC, with a sensitivity of 90.0% and a specificity of 76.7%. Finally, two promoters (RASSF1A and PGR-PROX) were informative for differentiating between OvCa vs. BOD, with a sensitivity of 80.0% and a specificity of 73.3%.

Conclusions

This proof-of-principle data show that differential methylation of promoters in cfpDNA may be a useful biomarker to differentiate between certain benign and malignant ovarian tumors.

Research Highlights

? Methylation patterns in cell-free DNA from blood can detect ovarian tumors ? These patterns are different in patients with benign and malignant tumors ? Methylation of blood DNA can be used for differential diagnosis of ovarian cancer     

Alveolar soft part sarcoma: A case report with emphasis on some unusual cytological features

Alveolar soft part sarcoma is a very rare, slow growing highly angiogenic tumor with poor prognosis. Most common site in children and infants is head and neck region and in adults it most commonly occurs in extremities especially thigh. In our case study, an 8 years old female patient presented with a gradually progressive left shoulder lump. FNAC from the lesion showed cellular smears with polyhedral and spindly cells showing abundant finely vacuolated cytoplasm, nuclear pleomorphism, intranuclear pseudoinclusions, and few bare nuclei. Perivascular arrangement of cells was peculiar in addition to the presence of intracytoplasmic metachromatic PAS positive diastase resistant granules. A presumptive diagnosis of alveolar soft part sarcoma with differentials of granular cell tumor and PEComa was considered and the lesion was excised. Although the histopathological features were not characteristic (ie, showing mainly solid pattern without classic alveolar pattern), immunohistochemistry were diagnostic (negative for S 100, Desmin, Cytokeratin, EMA, and moderate to strong nuclear positivity for TFE3). Thus, the diagnosis of alveolar soft part sarcoma was established. This case is being presented for its rarity and unique cytological and histopathological features.     

Bartonellosis and human immunodeficiency disease (AIDS): L-forms as persisters, activating factors, and mechanism of disease

Bartonella, genus Proteus, can cause immunodepressive disease. The organisms, in parasitized red blood cells, may invade the brain and every other system and space in the human body. Bartonella henselae is proposed to have a role in the pathogenesis of acquired immunodeficiency syndrome (AIDS) encephalopathy. Bartonella bacilliformis produces two known toxins that can induce spasm and angiomatosis, respectively, and manifest as diseases associated with symptomatic AIDS. The skin lesions of bartonellosis may be mistaken clinically and histologically for Kaposi's sarcoma. Bacteria of the genus Proteus produce L-forms: their elementary bodies may be mistaken for what are called the ‘human immunodeficiency viruses’ (HIV). Antibiotics, especially penicillin, induce bacteria to produce L-forms. Air pollution and high sugar, salt and fat diets are factors that may increase the lipid content of microbes that produce toxins and L-forms that may persist or revert to bacterial form.     

Azithromycin resistance and its molecular characteristics in Neisseria gonorrhoeae isolates from a tertiary care centre in North India

Treatment guidelines for management of uncomplicated gonorrhoeae have been recently modified owing to alarming upsurge in azithromycin resistance. This study investigated the prevalence and genetic determinants of gonococcal azithromycin resistance in India. Four (5.7%) of 70 gonococcal isolates were resistant to azithromycin. Of 16 isolates investigated for molecular mechanisms of resistance, 13 (81.3%) and 6 (37.5%) isolates exhibited mutations in coding and promoter regions of mtrR gene, respectively. However, ermA, ermB and ermC genes or mutations in rrl gene were absent in all isolates. Azithromycin resistance is low in India posing no immediate threat to use of dual-therapy for syndromic management.