'Hand inside glove': Useful method of burn dressing in children

The use of sterile surgical gloves in wound dressing is not new. It has been used previously in dressing of fresh wounds and in adjunct to the negative pressure wound management. Herein we describe an interesting case of burn wound dressing of hand in a child. Low cost, easy availability, better patient compliance and lesser chances of wound infection are special attributes of glove dressing

<span style="font-size: 20.0pt;mso-bidi-font-size:14.0pt;font-family:"Times New Roman","serif"">Evaluation of phytotoxicity and genotoxicity of uranyl nitrate in <i>Allium </i>assay system </span>

57-62<span style="font-size: 14.0pt;mso-bidi-font-size:8.0pt;font-family:" times="" new="" roman","serif""="">Uranyl nitrate inhibited root growth of <span style="font-size:15.0pt; mso-bidi-font-size:9.0pt;font-family:" times="" new="" roman","serif""="">Allium cepa at ≥ 25 <span style="font-size:15.0pt;mso-bidi-font-size:9.0pt; font-family:" times="" new="" roman","serif""="">µM <span style="font-size: 14.0pt;mso-bidi-font-size:8.0pt;font-family:" times="" new="" roman","serif""="">concentration. Fluorimetric analysis of metal uptake indicated the entry and accumulation of uranium into the root cell. Uranyl nitrate was neither clastogenic nor aneugenic as it failed to induce micronuclei significantly, but between 25 and 100<span style="font-size:15.0pt;mso-bidi-font-size:9.0pt;font-family: " times="" new="" roman","serif""="">µM <span style="font-size:14.0pt; mso-bidi-font-size:8.0pt;font-family:" times="" new="" roman","serif""="">concentration, it increased significantly the frequency of sister chromatid exchange over that of control, implying its genotoxicity that possibly interfered with DNA replication and <span style="font-size:14.0pt;mso-bidi-font-size: 8.0pt;font-family:" arial","sans-serif""="">/ <span style="font-size: 14.0pt;mso-bidi-font-size:8.0pt;font-family:" times="" new="" roman","serif""="">or repair process. </span

A Novel Anticancer Agent, 8-Methoxypyrimido4 `,5 `: 4,5]thieno(2,3-b) Quinoline-4(3H)-One Induces Neuro 2a Neuroblastoma Cell Death through p53-Dependent, Caspase-Dependent and Independent Apoptotic Pathways

Neuroblastoma is the most common cancer in infants and fourth most common cancer in children. Despite recent advances in cancer treatments, the prognosis of stage-IV neuroblastoma patients continues to be dismal which warrant new pharmacotherapy. A novel tetracyclic condensed quinoline compound, 8-methoxypyrimido 4 `,5 `: 4,5] thieno(2,3-b) quinoline-4(3H)-one (MPTQ) is a structural analogue of an anticancer drug ellipticine and has been reported to posses anticancer property. Study on MPTQ on neuroblastoma cells is very limited and mechanisms related to its cytotoxicity on neuroblastoma cells are completely unknown. Here, we evaluated the anticancer property of MPTQ on mouse neuro 2a and human SH-SY5Y neuroblastoma cells and investigated the mechanisms underlying MPTQ-mediated neuro 2a cell death. MPTQ-mediated neuro 2a and SH-SY5Y cell deaths were found to be dose and time dependent. Moreover, MPTQ induced cell death reached approximately 99.8% and 90% in neuro 2a and SH-SY5Y cells respectively. Nuclear oligonucleosomal DNA fragmentation and Terminal dUTP Nick End Labelling assays indicated MPTQ-mediated neuro 2a cell death involved apoptosis. MPTQ-mediated apoptosis is associated with increased phosphorylation of p53 at Ser15 and Ser20 which correlates with the hyperphosphorylation of Ataxia-Telangiectasia mutated protein (ATM). Immunocytochemical analysis demonstrated the increased level of Bax protein in MPTQ treated neuro 2a cells. MPTQ-mediated apoptosis is also associated with increased activation of caspase-9, -3 and -7 but not caspase-2 and -8. Furthermore, increased level of caspase-3 and cleaved Poly ( ADP Ribose) polymerase were observed in the nucleus of MPTQ treated neuro 2a cells, suggesting the involvement of caspase-dependent intrinsic but not extrinsic apoptotic pathway. Increased nuclear translocation of apoptosis inducing factor suggests additional involvement of caspase-independent apoptosis pathway in MPTQ treated neuro 2a cells. Collectively, MPTQ-induced neuro 2a cell death is mediated by ATM and p53 activation, and Bax-mediated activation of caspase-dependent and caspase-independent mitochondrial apoptosis pathways

Maternal Infection Is a Risk Factor for Early Childhood Infection in Filariasis

<div><p>Background</p><p>Global Program to Eliminate Lymphatic Filariasis (GPELF) launched by WHO aims to eliminate the disease by 2020. To achieve the goal annual mass drug administration (MDA) with diethylcarbamazine (DEC) plus albendazole (ABZ) has been introduced in all endemic countries. The current policy however excludes pregnant mothers and children below two years of age from MDA. Since pregnancy and early childhood are critical periods in determining the disease outcome in older age, the present study was undertaken to find out the influence of maternal filarial infection at the time of pregnancy on the susceptibility outcome of children born in a community after implementation of MDA for the first time.</p><p>Methodology and Principal Findings</p><p>The participants in this cohort consists of pregnant mothers and their subsequently born children living in eight adjacent villages endemic for filarial infections, in Khurda District, Odisha, India, where MDA has reduced microfilariae (Mf) rate from 12% to 0.34%. Infection status of mother and their children were assessed by detection of Mf as well as circulating filarial antigen (CFA) assay. The present study reveals a high rate of acquiring filarial infection by the children born to infected mother than uninfected mothers even though Mf rate has come down to < 1% after implementation of ten rounds of MDA.</p><p>Significance</p><p>To attain the target of eliminating lymphatic filariasis the current MDA programme should give emphasis on covering the women of child bearing age. Our study recommends incorporating supervised MDA to Adolescent Reproductive and Sexual Health Programme (ARSH) to make the adolescent girls free from infection by the time of pregnancy so as to achieve the goal.</p></div

The flow diagram of the cohort study.

<p>The flow diagram of the cohort study.</p

Status of filarial infection among the children born to filarial infected and uninfected mother in a MDA ongoing area of Odisha.

<p>*GM: Geometric mean</p><p>Status of filarial infection among the children born to filarial infected and uninfected mother in a MDA ongoing area of Odisha.</p

The flow diagram of the cohort study.

<p>The flow diagram of the cohort study.</p

MPTQ-mediated neuro 2a neuroblastoma cell death is associated with increased phosphorylation of ATM.

<p>A) 30 µM of MPTQ treated or untreated cells were fixed after 24 hours of treatment and immunolabelled with an antibody specific for phosphorylated ATM (ser1983). Detection was done using Alexa fluor 594 labelled secondary antibodies. Nuclei were stained by DAPI. Z-stack images were captured from multiple random fields, processed and displayed as described in methods. B) Intensity of phospho-ATM was measured in cytoplasmic and nuclear compartment. Histograms represent mean±standard deviation of three independent images from two independent experiments. C) Western blot analysis of phosphorylated ATM on cytosolic and nuclear fraction of MPTQ treated or untreated neuro 2a cells. Blots were also immunoblotted with anti-GAPDH and anti histone H3 antibodies for normalization. The results indicate increased phosphorylated ATM level in nuclear compartment of MPTQ treated cells than untreated cells. D) Densitometric analysis of phosphorylated ATM bands was performed and the values are plotted as mean±standard deviation of three independent isolates. Statistical analysis was made by Student’s t-test and p value is displayed. p value ≤0.05 is considered significant.</p

MPTQ induces nuclear DNA fragments of oligonucleosomal size in neuro 2a cells.

<p>A) Agarose gel electrophoresis of genomic DNA isolated from untreated and 2.5, 5, 10, 20 or 30 µM MPTQ treated cells for 48 hours indicated dose-dependent increased DNA fragmentation. B) Intensity of fragmented DNA was calculated and mean with standard deviation was plotted. Data represents three independent experiments. p value displayed for each duration of treatment was calculated by comparing with control sample using Student’s t-test. p value ≤5×10⁻² is considered significant. AU = Arbitrary Units.</p

MPTQ-mediated cell death is associated with activation of caspases of intrinsic apoptosis pathway but not of extrinsic pathway.

<p>A) Neuro 2a cells were cultured and treated with 30 µM of MPTQ for 24 hours and lysates were prepared. 60 µg of total proteins were resolved in 12% SDS-PAGE and immunoblotted with anti-caspase-8 or anti-caspase-2 or anti-caspase-9 or anti-caspase-3 or anti-caspase-7 antibody. Blots were stripped and immunoblotted with anti-GAPDH antibody. The results clearly indicate the activation of caspase-9, -3 and-7 but not caspase-8 and -2 in MPTQ treated cells. B) Immunocytochemistry of caspase-3 protein was performed as described earlier. Increased caspase-3 level was observed in the nucleus of MPTQ treated neuro 2a cells but not in control cells. C) Nuclear level of caspase-3 immunosignal was obtained using multi-cell scoring module and mean of three random images of two independent experiments were displayed as histograms. Error bar indicates standard deviation. D) Western blot analysis of cleaved caspase-3 level in cytosolic and nuclear fraction of MPTQ treated or untreated neuro 2a cells. Blots were also immunoblotted with anti-GAPDH and anti-histone H3 antibodies for normalization. E) Densitometric analysis of procaspase-3 and cleaved caspase-3 bands were made from cytosolic as well as from nuclear fractions. Cleaved caspase-3 to procaspase-3 ratio was obtained. Mean and standard deviation from three independent isolates were obtained and plotted as histograms. p value was calculated by Student’s t-test and is displayed which indicates significant increased mobilization of cleaved caspase-3 from cytoplasm to nucleus in MPTQ treated neuro 2a cells.</p