Multi-omics analysis identifies potential microbial and metabolite diagnostic biomarkers of bacterial vaginosis.

BACKGROUND: Bacterial vaginosis (BV) is a common clinical manifestation of a perturbed vaginal ecology associated with adverse sexual and reproductive health outcomes if left untreated. The existing diagnostic modalities are either cumbersome or require skilled expertise, warranting alternate tests. Application of machine-learning tools to heterogeneous and high-dimensional multi-omics datasets finds promising potential in data integration and may aid biomarker discovery. OBJECTIVES: The present study aimed to evaluate the potential of the microbiome and metabolome-derived biomarkers in BV diagnosis. Interpretable machine-learning algorithms were used to evaluate the utility of an integrated-omics-derived classification model. METHODS: Vaginal samples obtained from reproductive-age group women with (n = 40) and without BV (n = 40) were subjected to 16S rRNA amplicon sequencing and LC-MS-based metabolomics. The vaginal microbiome and metabolome were characterized, and machine-learning analysis was performed to build a classification model using biomarkers with the highest diagnostic accuracy. RESULTS: Microbiome-based diagnostic model exhibited a ROC-AUC (10-fold CV) of 0.84 ± 0.21 and accuracy of 0.79 ± 0.18, and important features were Aerococcus spp., Mycoplasma hominis, Sneathia spp., Lactobacillus spp., Prevotella spp., Gardnerella spp. and Fannyhessea vaginae. The metabolome-derived model displayed superior performance with a ROC-AUC of 0.97 ± 0.07 and an accuracy of 0.92 ± 0.08. Beta-leucine, methylimidazole acetaldehyde, dimethylethanolamine, L-arginine and beta cortol were among key predictive metabolites for BV. A predictive model combining both microbial and metabolite features exhibited a high ROC-AUC of 0.97 ± 0.07 and accuracy of 0.94 ± 0.08 with diagnostic performance only slightly superior to the metabolite-based model. CONCLUSION: Application of machine-learning tools to multi-omics datasets aid biomarker discovery with high predictive performance. Metabolome-derived classification models were observed to have superior diagnostic performance in predicting BV than microbiome-based biomarkers.

A Comparison of the Reliability of the Patellar Tendon-Trochlear Groove (PTTG) Distance and the Tibial Tuberosity-Trochlear Groove (TTTG) Distance Measured on MRI.

INTRODUCTION: An increased tibial tuberosity-trochlear groove (TTTG) distance is used for deciding a treatment plan in patello-femoral instability (PFI). The centre of the patellar tendon and the chondral trochlear groove can be directly visualised on MRI, and measured, giving the patellar tendon-trochlear groove (PTTG) distance. A study was designed to compare the inter-rater and the test-retest reliabilities of PTTG and TTTG measurements in MRI of patients without PFI and in a group with PFI. MATERIALS AND METHODS: This cross-sectional reliability study was done on archival MRI films of 50 patients without patellar instability and 20 patients with patellar instability. TTTG and PTTG distances were independently measured by two orthopaedic surgeons and two radiologists. A hybrid PTTG measurement with bony landmarks on the femoral side and the patellar tendon landmark on the tibial side, was used to estimate the influence of the differences in the femoral and tibial landmarks on the difference in reliabilities. The intra-class correlation coefficient (ICC) was calculated for all four raters, as well as separately for each rater. RESULTS: The PTTG distance had a higher inter-rater reliability (ICC=0.86, 95% CI=0.79-0.92) compared to the TTTG distance (ICC=0.70, 95% CI=0.59-0.80) in patients without PFI. Similar trends were seen in patients with PFI (0.83 vs 0.66). The inter-rater reliability for the hybrid PTTG distance was found to lie in between the TTTG and PTTG. CONCLUSIONS: The MRI-based PTTG distance had better inter-rater reliability compared with the MRI-based TTTG distance.

CDK4 in lung, and head and neck cancers in old age: evaluation as a biomarker.

BACKGROUND: Cyclin dependent kinases (CDK) are key factors in promoting the initiation and development of tumors. These kinases are important for maintenance of mitochondrial biogenesis and imbalance in their expression in old age may lead to the oxidative stress. Lung cancer (LC), and head and neck squamous cell carcinoma (HNSCC) are two very prominent cancers in older Indians. Both the cancers are showing increasing trend in older population. The present study assessed serum concentration of one of the kinases; CDK4 in older LC and HNSCC patients. METHODS: The study included 100 subjects each of LC and HNSCC; and older subjects without cancer or any major health problems as controls. Serum CDK4 concentration was estimated using real-time label-free Surface plasmon resonance (SPR) and was verified by western blot. RESULTS: Significant elevation in serum CDK4 was observed in cases with LC and HNSCC compared to controls. HNSCC patients with higher CDK4 expression had distinctly shorter survival than patients with comparatively lower CDK4 expression. No such difference was observed in LC patients. The germ line mutation study of this gene in Exon-2 was performed and none was observed among cases and controls. CONCLUSION: It can be concluded that older patients with HNSCC and lung cancer have raised serums CDK4 levels, which has the potential to emerge as a biomarker in clinical practice.

Aspiration pneumonia related deaths in head and neck cancer patients: a retrospective analysis of risk factors from a tertiary care centre in North India.

BACKGROUND: Aspiration pneumonia is an important cause of death in head and neck cancer patients. This study therefore aimed to evaluate the risk factors associated with aspiration pneumonia in head and neck cancer patients. METHODS: Hospital death records from 12 years (2000-2012) were reviewed to obtain the number of deaths. Treatment details and cause of death were analysed. Statistical analysis was performed to identify the risk factors for aspiration pneumonia. RESULTS: The records revealed that aspiration pneumonia was the cause of death in 51 out of 85 patients. Primary tumour site (oropharynx and hypopharynx, odds ratio 3.3; 95 per cent confidence interval 1.17-9.4, p = 0.02) and advanced tumour stage (odds ratio 4.2, 95 per cent confidence interval 1.16-15.61, p = 0.02) had significant negative impacts on aspiration pneumonia related mortality. CONCLUSION: Advanced pharyngeal cancer patients are at an increased risk of aspiration pneumonia related death. Investigations for the early detection of this condition are recommended in these high-risk patients.

Correlation between lead and cadmium concentration and semen quality.

There are contrary reports of association of lead and cadmium with the decline in semen quality. This study evaluates whether seminal lead (Pb) and cadmium (Cd) at environmental concentration are associated with altered semen quality. We conducted a study of healthy fertile and infertile men 20-43 years of age attending the Andrology Laboratory of Reproductive Biology Department for semen analysis. The semen analysis was carried out according to the WHO 2010 guidelines. Seminal lead and cadmium were estimated by ICP-AES. The lead and cadmium values were significantly higher in infertile subjects. A negative association between seminal lead or cadmium concentration and sperm concentration, sperm motility and per cent abnormal spermatozoa was found. This study shows that exposure to Pb (5.29-7.25 µg dl(-1) ) and cadmium (4.07-5.92 µg dl(-1) ) might affect semen profile in men. Age, diet, smoking and tobacco chewing habits may have an influence on the increase in exposure to Pb and Cd in the individual subjects.

Reproductive toxicity of lead, cadmium, and phthalate exposure in men.

Environmental toxicants viz lead or cadmium and phthalate esters (di(2-ethylhexyl) phthalate [DEHP], dibutyl phthalate [DBP], and diethyl phthalate [DEP]) widely found in different environmental strata are linked to deteriorating male reproductive health. The objective was to assess the relationships between the seminal lead, cadmium, and phthalate (DEHP, DBP, DEP) concentrations at environmental level and serum hormone levels and semen quality in non-occupationally exposed men and specify the effect of individual and combined exposure of toxicants on semen quality. A study of 60 male partners of couples attending the Andrology Laboratory of the Reproductive Biology Department, All India Institute of Medical Sciences (AIIMS), New Delhi, India for semen analysis to assess their inability to achieve a pregnancy was selected for the study. The results of univariate and stepwise multiple regression analysis in the unadjusted model showed a significant correlation between lead or cadmium and phthalates DEHP/DBP/DEP and sperm motility, sperm concentration, and DNA damage. After adjusting for potential confounders, an association with lead or DEHP was only observed. The present data shows that lead (Pb) or cadmium (Cd) or phthalates might independently contribute to decline in semen quality and induce DNA damage. Phthalates might influence reproductive hormone testosterone. These findings are significant in light of the fact that men are exposed to a volley of chemicals; however, due to the small sample size, our finding needs to be confirmed in a larger population.

Association between environmental exposure to p, p'-DDE and lindane and semen quality.

Scientific concern exists about the toxic effect of dichlorodiphenyldichloroethylene (p, p'-DDE) and lindane on male infertility, and the mechanism underlying male reproductive toxicity of this pesticide remains unanswered. We investigated not only the possible association between the chlorinated pesticide levels and semen quality in nonoccupationally exposed men, but also the probable mode of action using mitochondrial membrane potential (MMP), reactive oxygen species (ROS), lipid peroxidation (LPO), and sperm chromatin structure assay (SCSA). A study in 278 men (21-40 years old) who visited Obstetrics and Gynecology Department, KGMU, Lucknow, for semen analysis was conducted. We performed semen analysis according to the WHO guidelines, while p, p'-DDE and lindane analysis was done by the GLC and LPO by the spectrophotometer, and the sperm mitochondrial status, ROS, and SCSA with the flow cytometer. The questionnaire data showed no significant difference in the demographic characteristics between the two groups, i.e., trying to conceive >1 year and proven fertility. However, a significant difference in the concentration of p, p'-DDE and lindane was observed between the groups. When the subjects were divided among four categories by quartile of exposure, the subjects in the highest quartile showed low sperm motility as compared to the subjects in the lowest quartile. Pearson's correlation showed a significant negative correlation between semen p, p'-DDE, lindane level, and sperm quality and positive association with the number of cells with depolarized mitochondria, elevation in ROS production and LPO, and DNA fragmentation index (DFI). The findings are suggestive that these toxicants might cause a decline in semen quality, and these effects might be ROS, LPO, and mitochondrial dysfunction mediated.

Percentage body fat in apparently healthy school children from northern India.

CONTEXT: Increased prevalence of obesity in childhood and adolescence, defined by the use of body mass index (BMI), has drawn attention towards direct measurement of body fat. OBJECTIVE: To develop age-and sex-specific reference distribution of body fat in apparently healthy North-Indian children in the age group of 7-17 years and to assess agreement between obesity (defined by BMI) and excess body fat. DESIGN: Study subjects for this cross sectional study included 1640 apparently healthy school children (825 boys; 815 girls) aged 7-17 years. Total body fat was measured by dual energy X-rays absorptiometry (DXA). The excess body fat by DXA was defined by two methods, prevalence matching and with the use of 85th and 95th centile cutoffs. RESULTS: The mean ± SD, 3rd, 10th, 25th, 50th, 75th, 90th and 97th centile values of percentage body fat (PBF) are presented. PBF was highly correlated with BMI in both boys and girls (all boys: r=0.76, P<0.0001; all girls r=0.81, P<0.0001). There was no significant difference noted in PBF between boys and girls at the age of 7-8 years. From 9 years onwards, girls had significantly higher PBF than boys. Moderate degree of agreement was observed between BMI and PBF by DXA by both methods. CONCLUSIONS: Smoothened reference distribution of PBF for North-Indian children and adolescents in Delhi are provided. Indian children accumulate more body fat during peri-pubertal years in comparison with US children.

Relative efficacy of weekly and two differing doses of daily iron-folate supplementation in improving hemoglobin in mild and moderately anemic children between 3 and 5 years of age: a cluster randomized trial.

BACKGROUND/OBJECTIVES: In India, 75% of children <5 years of age have anemia. The National Nutritional Anemia Control Program (NNACP) recommends 20 mg iron and 100 µg folic acid (IFA) supplementation for 100 days/year, but still anemia prevalence has remained high. To accelerate the progress, suggestions include increase in IFA to therapeutic dose or supervised weekly supplementation to improve compliance. The objectives of this study was to compare the hemoglobin response with two dosages of daily (20 mg iron and 100 µg folic acid, or 40 mg iron and 200 µg folic acid) and weekly (40 mg iron and 200 µg folic acid) IFA supplementation in children of 3-5 years of age with mild or moderate anemia (hemoglobin 7-10 g/dl). SUBJECTS/METHODS: Community-based cluster randomized control trial in nine adjoining Anganwadi Centers. Four hundred twenty six enrolled participants received directly supervised IFA tablet supplementation as per the above three groups. After 100 days, the number of available subjects in the NNACP daily dose (A), daily dose doubled (B) and weekly dose (C) groups were 112, 114 and 110, respectively. Hemoglobin was estimated at baseline, 50 and 100 days by the Cynmeth hemoglobin method. RESULTS: At 50 days, there were no differences between the three groups, but at 100 days, adjusted hemoglobin was lowered with weekly supplementation. The mean (95% confidence interval) hemoglobin (g/dl) differences were: (i) A-B: -0.05 (-0.17, 0.05), (ii) A-C: -0.38 (-0.50, -0.27) and (iii) B-C: -0.33, (-0.45, -0.21). Anemia reduction was 18.8%, 18.4% and 10.9%, respectively, in the three groups. CONCLUSION: Directly supervised IFA supplementation at the NNACP or double dose is equally efficacious but superior to weekly regimen.

Hearing status in children with frequently relapsing and steroid resistant nephrotic syndrome.

BACKGROUND: Children with idiopathic nephrotic syndrome (INS) are at risk of hearing impairment due to nephrotoxic drugs and biochemical impairments. METHODS: Forty children with INS aged 5-16 years [20 patients with frequently relapsing nephrotic syndrome (FRNS)/steroid dependent nephrotic syndrome (SDNS) and 20 with steroid resistant nephrotic syndrome (SRNS)] and 20 normal healthy controls were enrolled in this study. Pure tone audiometry was done using the ALPS AD 2000 audiometer. Sensorineural hearing loss was diagnosed when the bone conduction level was >20 dB and the difference in air to bone gap was ≤15 dB. Based on the air conduction (AC) threshold, deafness was graded into the following categories: mild (26-40 dB), moderate (41-55 dB), moderately severe (56-70 dB), severe (71-91 dB) and profound (>91 dB). RESULTS: Children with FRNS/SDNS had a higher threshold for hearing at frequencies of 250 and 500 Hz, respectively, than the controls. Of the children in the FRNS/SDNS category, three (15 %) had mild sensorineural hearing impairment. These children had a low serum calcium level (P < 0.03) and received higher cumulative doses of furosemide (P < 0.04). Children with SRNS had a higher threshold for hearing at frequencies of 250, 500, 1,000, and 2,000 Hz, respectively, than the controls. Of the 20 children with SRNS, ten (50 %) had sensoineural hearing impairment (8 mild, 2 moderate). Children with SRNS with a hearing defect had received a higher cumulative dose of furosemide (P < 0.03). CONCLUSIONS: Children with FRNS/SDNS and SRNS are at risk of sensorineural hearing impairment. The risk factors associated with this impairment were higher cumulative doses of furosemide and hypocalcemia. Larger prospective cohort studies are required to evaluate this association.