Parental perception of child vulnerability among mothers of very low birth weight infants: psychological predictors and neurodevelopmental sequelae at 2 years.

OBJECTIVE: The objective of this study was to examine the impact of maternal psychological distress on the development of parental perception of child vulnerability (PPCV) in mothers of very low birth weight (VLBW) infants; and to examine the impact of PPCV on neurodevelopmental outcome in VLBW infants in the second year of life. STUDY DESIGN: This is a prospective study of 69 mothers and their VLBW infants recruited from 2011 to 2012 for whom maternal psychological data were collected during the neonatal intensive care unit (NICU) hospitalization. Maternal PPCV was assessed at 4 months corrected age (CA). Neurodevelopmental outcome was assessed at 20 months CA. Regression analyses modeled the development of PPCV and the impact of PPCV on neurodevelopmental outcome. RESULTS: PPCV at 4 months CA was predicted by maternal anxiety and history of previous fetal loss reported during the NICU stay. Higher PPCV at 4 months CA was associated with lower language scores at 20 months CA. CONCLUSION: Targeted interventions aimed at reducing PPCV in the NICU are supported.

Neurodevelopmental impact of hydrocortisone exposure in extremely low birth weight infants: outcomes at 1 and 2 years.

OBJECTIVE: Postnatal steroids are used in neonatal intensive care units despite known side effects. Hydrocortisone (HC) use persists as it is believed to have less deleterious effects on neurodevelopmental (ND) outcome compared to other steroids. The literature is sparse with respect to the ND impact of HC use in recent years. Hence, we sought to examine the effect of HC use on ND outcome in a contemporary cohort of extremely low birth weight (ELBW) infants. STUDY DESIGN: A total of 175 ELBW infants (86 HC exposed, 89 steroid naive) born in 2008 to 2010 were compared for mortality, morbidity and ND outcome at 8 and 20 months corrected age. Outcome measures included neurologic exam and results of the Bayley Scales of Infant and Toddler Development-III (BSITD-III). Multiple regression analyses adjusted for the effect of other risk factors on outcome. RESULT: Overall, 65 (75%) of the HC and 74 (83%) of the no-HC groups survived to discharge. HC infants were smaller (mean birth weight (BW) 719 ± 127 g vs 837 ± 99 g) and of lower gestational age (GA) (mean GA 26.0 ± 1.7 weeks vs 27.5 ± 1.8 weeks) compared to the no-HC group. Patients in the HC group were more likely to be a multiple, have a severely abnormal head ultrasound, bronchopulmonary dysplasia, retinopathy of prematurity, necrotizing enterocolitis and receive treatment for patent ductus arteriosus and hypotension than those in the no-HC group. Of the HC group, the mean age at treatment was 20 ± 19 days, mean duration of treatment 49 ± 37 days. At 8 months, the HC group had lower mean motor (87 ± 18 vs 95 ± 15, P = 0.028) and fine motor (9 ± 2.9 vs 10.5 ± 2.6, P = 0.005) and higher rate of subnormal motor (44 vs 15%, P = 0.002) and fine motor scores (24 vs 6.5%, P = 0.017). In regression analyses, HC exposure >7 days was significantly related to worse outcome on fine motor scores at 8 months while cumulative days of HC exposure was a predictor of worse outcome on language at 8 months and motor outcome at 20 months. Each additional day of HC exposure increased the odds of subnormal receptive and expressive language in the first year of life by 4 and 2%, respectively, and increased odds of subnormal motor function by 2% in the 2nd year of life. CONCLUSION: HC exposure for >7 days is associated with worse performance in fine motor skills in the first year of life, while cumulative HC exposure negatively impacts receptive and expressive language skills in the first year and motor skills in the second year of life after adjusting for neonatal and social risk factors.

Cardiorespiratory events in extremely low birth weight infants: neurodevelopmental outcome at 1 and 2 years.

OBJECTIVE: To examine the association between cardiorespiratory events (CRE) and neurodevelopmental (ND) outcome at 8 and 20 months corrected age (CA) in a contemporary extremely low birth weight (ELBW )cohort. STUDY DESIGN: Retrospective chart review of 98 ELBW infants born in 2009 to 2010 who completed ND assessments at 8 and 20 months CA. Neonatal, sociodemographic, CRE and ND data were collected. ND outcome measures included neurologic examination and results from the Bayley Scales of Infant and Toddler Development-III. Multiple regression analyses adjusted for the impact of neonatal risk factors on ND outcome. RESULT: After adjusting for neonatal and social variables, greater frequency of CRE was related to worse language scores at 8 months, while CRE of greater severity were related to worse language at 20 months CA. CONCLUSION: CRE in ELBW infants have impact on language development in the first two years of life.

The synaptic protein encoded by the gene Slc10A4 suppresses epileptiform activity and regulates sensitivity to cholinergic chemoconvulsants.

The expanding number of disease-causing dysfunctions of synaptic proteins illustrates the importance of investigating newly discovered proteins involved in neuronal transmission. The gene Slc10A4 encodes a recently described carrier protein present in pre-synaptic terminals of cholinergic and monoaminergic neurons. The biological significance of this recently described transporter protein is currently unknown. We here investigated whether absence of the Slc10a4 protein has any impact on function of the cholinergic system. We first investigated the sensitivity of Slc10a4 null mice to cholinergic stimulus in vitro. In contrast to wild type mice, gamma oscillations occurred spontaneously in hippocampal slices from Slc10a4 null mice. Furthermore, moderate treatment of Slc10a4 null slices with the cholinergic agonist carbachol induced epileptiform activity. In vivo, 3-channel EEG measurements in freely behaving mice revealed that Slc10a4 null mice had frequent epileptiform spike-activity before treatment, and developed epileptic seizures, detected by EEG and accompanied by observable behavioral components, more rapidly after injection of the cholinergic agonist pilocarpine. Similar results were obtained on non-operated mice, as evaluated by behavioral seizures and post mortem c-Fos immunohistochemistry. Importantly, Slc10a4 null mice and wild type control mice were equally sensitive to the glutamatergic chemoconvulsant kainic acid, demonstrating that absence of Slc10a4 led to a selective cholinergic hypersensitivity. In summary, we report that absence of the recently discovered synaptic vesicle protein Slc10a4 results in increased sensitivity to cholinergic stimulation.

Analysis of grains grown on fly ash treated soils.

One time application of fly ash was done at the rate of 200 t/hectare in the crop lands in Odisha, India. The increase in yield of maize grains was 28% and 34% over the control values whereas the increase in yield of rice grains was 40% and 13% at Malud and Dhenkanal, respectively. The results indicated that fly ash is an amendment for soil which can improve the physical and chemical properties of deficient soil and improves the soil fertility and crop yield.

Elemental analysis of coal and coal ASH by PIXE technique.

Coal and coal ash samples were characterized by particle induced X-ray emission spectroscopic technique. Sixteen elements, namely K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Ga, As, Rb, Sr, Y and Pb were quantified in this study. Elements like K, Ca, Ti and Fe were present as major elements, whereas, other elements like V, Cr, Mn, Co, Ni, Cu, Zn, As, Se, Rb, Sr and Pb were present in trace level. The enrichment ratio of different ash samples with respect to coal were also estimated and discussed.

A randomised comparative study between low-dose intravenous magnesium sulphate and standard intramuscular regimen for treatment of eclampsia.

In a randomised control study, we evaluated the efficacy of intravenous low-dose magnesium sulphate for the management of eclampsia. A total of 144 women with eclampsia were divided into a study group and a control group of 72 women each. The study group received 0.75 g/h of magnesium sulphate intravenously after a loading intravenous dose of 4 g and the control group was given the standard intramuscular regimen as advocated by Pritchard. The primary outcome measure was recurrence rate of the seizures. The secondary outcome measures were development of magnesium toxicity if any, and maternal and perinatal outcomes. The difference in the incidence of fit recurrence was statistically insignificant when both groups were compared (7.46% vs 8.57%, p = 0.939). The total dose of magnesium sulphate was significantly lower in the intravenous group (p < 0.0001), in which no patient developed magnesium toxicity. Low-dose intravenous magnesium sulphate was found to be as effective as the standard intramuscular regimen, while maintaining a high safety margin.

Effectiveness and safety of neuraminidase inhibitors in reducing influenza complications: a meta-analysis of randomized controlled trials.

BACKGROUND: Several studies suggest that neuraminidase inhibitors (NIs) can reduce the duration of influenza symptoms. However, data regarding their effectiveness in reducing influenza complications are scarce. METHODS: We evaluated the effectiveness of NIs in reducing influenza complications and mortality of patients with seasonal influenza, by performing a meta-analysis of randomized controlled trials (RCTs), retrieved from PubMed, Cochrane Central Register of Controlled Trials and Scopus databases, comparing NIs with placebo. RESULTS: Eleven RCTs (10 double-blind) were included; 8 involved adults/adolescents. In total, 5315 patients were included; 3491 (65.7%) with confirmed infection. Total influenza-related complications were significantly less likely in otherwise healthy patients with confirmed influenza infection that were treated with NIs versus placebo [7 RCTs, 2621 patients, risk ratio (RR) = 0.74, 95% confidence interval (CI) = 0.58-0.95] This finding was more pronounced in high-risk patients [4 RCTs, 475 patients, RR = 0.37, 95% CI = 0.24-0.59]; P < 0.01 for the chi(2) test for subgroup differences. In the comparisons regarding individual complications, a trend in favour of NIs was observed. Acute otitis media was significantly less likely in patients with confirmed influenza infection treated with NIs versus placebo (3 RCTs, 1124 patients, RR = 0.50, 95% CI = 0.30-0.85). No differences were found in the comparisons regarding the safety outcomes. No deaths were observed in trials that reported on mortality. CONCLUSIONS: NIs seem to be effective in reducing total influenza-related complications in otherwise healthy and high-risk patients, and have an acceptable safety profile. However, RCTs providing separate data for mild to serious complications and detailed reporting of adverse events and mortality are needed.

Prevalence and significance of antiphospholipid antibodies in selected at-risk obstetrics cases: a comparative prospective study.

In a prospective comparative study we screened 112 women with a past history either of pre-eclampsia, eclampsia, recurrent abortion, IUGR, IUFD or abruptio placentae, with no apparent aetiology and a demographically matched cohort of 106 women having a past history of uncomplicated pregnancy outcome for the presence of antiphospholipid antibodies (aPL) and their significance. In the former group, the prevalence of aPL ranged from 10-46.87% compared with 8.49% in the later group. In women with the presence of aPL, the incidence of pre-eclampsia, early onset pre-eclampsia and abruptio placentae were 25%, 14.58% and 18.75%, respectively. In the same group, the abortion rate was 25% and live-birth rate was 64.58% with IUFD rate of 10.42%. Fetal morbidity rates were also higher in the mothers with aPL positivity, the incidence of IUGR was 27.08% and oligohydramnios was 33.33% in them. All these complications were statistically significant when compared with those of aPL negative mothers.

RT-PCR assay for the detection of infective (L3) larvae of lymphatic filarial parasite, Wuchereria bancrofti, in vector mosquito Culex quinquefasciatus.

BACKGROUND & OBJECTIVES: Periodic monitoring of vector population for infection and infectivity rates is central to the evaluation of the filariasis elimination strategies in endemic areas to monitor the success of MDA and also to establish endpoints for intervention. The main objective of this study was to develop a RT-PCR assay, based on L3 stage-specific primers to detect the presence of infective stage larvae of filarial parasite, Wuchereria bancrofti in the vector Culex quinquefasciatus. MATERIAL & METHODS: Subtracted probe development technique was employed for the identification of infective stage (L3) specific genes. The subtracted cDNA was labeled by non-radioisotopic method and used for screening cDNA library of L3 stage larvae of W. bancrofti constructed in UniZap XR. Recombinants were probed and identified from the library. The inserts of the recombinant clones were purified and sequenced. Primers were designed based on the sequence information of three recombinant clones for detecting L3 larvae of W. bancrofti in the vector by RT-PCR assay. Preliminary laboratory evaluation was carried out to assess the sensitivity and specificity of WbL31 RT-PCR assay. RESULTS: cDNA library of L3 stage of W. bancrofti constructed in UniZap XR vector, constituted 5 x 10(5) phages with 80-90% recombinant phages and the size of inserts varied from 0.1 to 1.0 kb. When subtracted cDNA was random prime labeled and used for screening cDNA library of L3 stage of W. bancrofti constructed in UniZap XR, 18 clones were identified from the library. Three genes were found up-regulated in the L3 stage, out of which WbL31 (cuticular collagen) was found to be useful in detecting L3 larvae of W. bancrofti in the vector by RT-PCR assay with high specificity and sensitivity (98-100%). CONCLUSION: Present paper marks first report on the development of an infective stage-specific RT-PCR assay (WbL31 RT-PCR assay) to detect L3 stage W. bancrofti in the vector. This assay will have potential application in assessing the transmission of infection and hence in decision-making related to elimination programme.

Exposure to quinolones is associated with carbapenem resistance among colistin-susceptible Acinetobacter baumannii blood isolates.

In this study, we explored risk factors associated with bacteraemia caused by colistin-susceptible/carbapenem-resistant (Co(S)/Ca(R)) Acinetobacter baumannii. A retrospective cohort study of hospitalised patients with A. baumannii bacteraemia was performed at a tertiary care hospital over a 44-month period. Thirty-nine patients with bacteraemia due to A. baumannii (35 Intensive Care Unit and 4 ward patients) were included in the analysis. Twenty-five patients (64%) had bacteraemia due to Co(S)/Ca(R)A. baumannii and 14 patients (36%) had bacteraemia due to colistin-susceptible/carbapenem-susceptible A. baumannii. Mortality was 56% (14/25) and 35.7% (5/14) for patients in the two groups, respectively (P=0.22). Bivariate analysis showed that prior exposure to fluoroquinolones (P=0.01) and antipseudomonal penicillins (P=0.004) as well as a higher number of antibiotics in use on the day of bacteraemia (P=0.02) were associated with isolation of a Co(S)/Ca(R) strain among patients with A. baumannii bacteraemia. Multivariate analysis using a backward logistic regression model showed that only exposure to fluoroquinolones was associated with development of Co(S)/Ca(R)A. baumannii bacteraemia (odds ratio=11.6; 95% confidence interval 2.4-55.9; P=0.02). The appearance of Co(S)/Ca(R)A. baumannii infections represents a major threat to critically ill hospitalised patients. Exposure to fluoroquinolones is an independent risk factor for development of Co(S)/Ca(R)A. baumannii bacteraemia.

Identification of a molecular marker for genotyping human lymphatic filarial nematode parasite Wuchereria bancrofti.

In India, Mass Drug Administration is on going towards elimination of lymphatic filariasis in many areas, which might lead to intense selection pressure on the parasite populations and their genetic restructuring. This calls for molecular finger printing of Wuchereria bancrofti parasite populations at national level and monitoring genetic changes in the future. For this purpose a reliable, less expensive, rapid, and reproducible molecular tool is necessary, which is not available for W. bancrofti at this time. We identified robust molecular markers based on the comparison of random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) profiles and the genetic data generated from parasite populations collected from areas in Northern (Varanasi, Uttar Pradesh state), Southern (Kozhikode, Kerala State) and Central regions (Jagdalpur, Chattisgarh state) of India, where lymphatic filariasis is endemic for many decades. RAPD profiles for these parasite populations were generated using three different primers and the dendrograms constructed using the profiles were all different. In order to identify appropriate RAPD primer(s), we compared the results of RAPD with the fingerprint profile and genetic data obtained by the more reliable AFLP technique, using the parasite populations from the same areas. RAPD marker (OP8) primer produced phylogenetic data almost similar to that of AFLP analysis. The marker was able to reveal variations between the parasite populations collected from Varanasi, Kozhikode, and Jagdalpur. Most importantly, RAPD primer OP8 produced reproducible results, when tested in three different trials. In view of the limited availability of W. bancrofti parasite DNA, along with a lower cost and ease of performance, RAPD appears to be more suitable compared to AFLP at the present juncture, since complete genome information of this parasite is still not available. Thus, RAPD primer OP8 can be a very useful molecular maker for DNA finger printing of W. bancrofti populations at present.

Polymorphism of gp15/400 allergen gene of W. bancrofti from different regions of India endemic for lymphatic filariasis.

Nematode polyprotein allergens (NPA) are lipid binding/transport molecules that elicit elevated levels of IgE response in the infected host, leading to Th2 type of immune response. They also transport arachidonic acid and its metabolites that are known to be involved in the action of antifilarial drug, Diethylcarbamazine and hence are of great significance for the control of lymphatic filariasis. We investigated the polymorphism of gp15/400 polyprotein of 35 isolates of lymphatic filarial parasite Wuchereria bancrofti collected from different geographic locations of India. The repeat sub-unit of the gene was found to be highly conserved in all the isolates with only two nucleotide synonymous changes at positions 286 (A-G) and 337 (C-T). Since this molecule is highly conserved and has multifarious roles in the survival and pathogenesis of the parasite it has good potential as a target for drug, immunodulation tool and immunotherapy development.

Polymorphism of gp15/400 allergen gene of Wuchereria bancrofti from different regions of India endemic for lymphatic filariasis.

Nematode polyprotein allergens (NPA) are lipid-binding/transport molecules that elicit elevated levels of IgE response in the infected host, leading to Th2 type of immune response. They also transport arachidonic acid and metabolites that is known to be the action of antifilarial drug, diethylcarbamazine, and hence are of great significance to the control of lymphatic filariasis. We investigated the polymorphism of gp15/400 polyprotein of 35 isolates of lymphatic filarial parasite Wuchereria bancrofti collected from different geographic locations of India. The repeat subunit of the gene was found to be highly conserved in all the isolates with only two nucleotide synonymous changes at positions 369 (A-G) and 375 (C-T). As this molecule is highly conserved and has multifarious roles in the survival and pathogenesis of the parasite, it has good potential as a target for drug, immunodulation tools and immunotherapy development.

The diversity of definitions of multidrug-resistant (MDR) and pandrug-resistant (PDR) Acinetobacter baumannii and Pseudomonas aeruginosa.

Different definitions of the terms multidrug-resistant (MDR) and pandrug-resistant (PDR) Acinetobacter baumannii and Pseudomonas aeruginosa have been used in the biomedical literature. The authors searched for relevant studies indexed in the PubMed database (01/2000-09/2005) to systematically examine the various definitions of MDR and PDR for these bacteria. Initially 107 retrieved relevant studies were reviewed. Ninety-two studies were further analysed, 50 of which focused on A. baumannii and 42 on P. aeruginosa. A considerable diversity of definitions of the terms MDR and PDR A. baumannii and P. aeruginosa was found. Of note, the term PDR was inappropriately used in all five studies that used it. The review reveals that various definitions have been used for the terms MDR and PDR A. baumannii and P. aeruginosa, a fact that causes confusion to researchers and clinicians. The authors believe that at least a widely accepted definition for PDR A. baumannii and P. aeruginosa should be uniformly used worldwide.

Risk factors for isolation of strains susceptible only to polymyxin among patients with Pseudomonas aeruginosa bacteremia.

We conducted a case-control study to identify risk factors associated with the isolation of Pseudomonas aeruginosa strains susceptible only to polymyxin from blood by comparing data between 16 patients with blood isolates that were susceptible only to polymyxins and 40 patients with blood isolates that were susceptible to carbapenems. The multivariable analysis showed that exposure to carbapenems was associated with the development of P. aeruginosa bacteremia susceptible only to polymyxin (odds ratio, 9.0; 95% confidence interval, 2.4 to 34.3; P = 0.001).

Genetic variability of the human filarial parasite, Wuchereria bancrofti in South India.

The genetic variability of the lymphatic filarial parasite Wuchereria bancrofti, from three localities (one urban and two rural areas) in southern India, endemic for filariasis was studied using random amplified polymorphic DNA (RAPD) markers. The RAPD profiles were generated for 21 parasite populations (7 populations from each area), using a 10-mer random primer. The analysis of profiles indicated the existence of considerable genetic variability among parasite populations. The Nei's gene diversity between the individual populations in the 2 areas (one urban and another rural) was comparatively greater (0.3372+/-0.1462 & 0.2830+/-0.1764) than that of populations in another village (0.0490+/-0.1373). The greater genetic diversity among the former areas may be due to human migration, endemicity for long time and drug (diethyl-carbamazine citrate) pressure unlike the populations of latter village where the filariasis is relatively a recent introduction and which was never under active chemotherapy. The Nei's genetic distance was estimated and the phylogenetic tree was constructed using 'UPGMA'. These analyses indicated the prevalence of at least two genetically distinct clusters, among the populations studied, their maximum genetic distance being 0.2444. The finding of two genetic 'variants' of W. bancrofti, in the present study, may have important implications in filariasis epidemiology and control/elimination programmes.

A rapid and simplified method of DNA extraction for the detection of Brugia malayi infection in mosquitoes by PCR assay.

Currently used protocols for the extraction of filarial parasite DNA from mosquito samples are tedious and involve extensive use of expensive and hazardous chemicals. Therefore, in order to arrive at a simple procedure, four different methods (A, B, C and D) were tried for the extraction of DNA from mosquitoes infected with filarial parasite, Brugia malayi. Method D was found to be as efficient as the current procedure for the extraction of DNA from a single microfilaria in pools of 25 mosquitoes and the DNA was suitable for polymerase chain reaction (PCR) amplification, yielding a band of 322 base pairs with primers specific for B. malayi. Method D involved drying and crushing the mosquitoes to a powder, which was homogenized in 100 microl TE buffer, vortexed, boiled for 10 min, centrifuged at 14000 r.p.m. for 10 min, and the supernatant used for the PCR assay. Dot-blot hybridization confirmed the specificity of the PCR amplified fragment. The DNA extracted by this method was stable for about 1 year. When comparing with the standard method, the cost of a single PCR reaction, inclusive of DNA extraction, was reduced by 50% and the hands on time was minimized fivefold. Hence, this simple TE-based method is rapid, safe and also cost-effective in assessing the B. malayi infection in pools of vector mosquitoes.