The p53 and Calcium Regulated Actin Rearrangement in Model Cells.

Long-term cellular stress maintains high intracellular Ca2+ concentrations which ultimately initiates apoptosis. Our interest is focused on how the gelsolin (GSN) and junctional mediating and regulating Y protein (JMY) play important roles in stress response. Both of these proteins can bind p53 and actin. We investigated using in vitro fluorescence spectroscopy and found that the p53 competes with actin in GSN to inhibit p53-JMY complex formation. A high Ca2+ level initializes p53 dimerization; the dimer competes with actin on JMY, which can lead to p53-JMY cotransport into the nucleus. Here we investigated how the motility and division rate of HeLa cells changes due to low-voltage electroporation of GSN or JMY in scratching assays. We revealed that JMY inhibits their motion, but that it can accelerate the cell division. GSN treatment slows down cell division but does not affect cell motility. HeLa cells fully recovered the gap 20 h after the electroporation with JMY and then started to release from the glass slides. Taken together, our in vitro results indicate that GSN and JMY may play an important role in the cellular stress response.

Evaluation of p21 expression and related autism-like behavior in Bisphenol-A exposed offspring of Wistar albino rats.

BACKGROUND: Bisphenol A (BPA), an endocrine disruptor, may be involved in the etiology of autism spectrum disorders (ASD); however, the mechanism of neuronal and astrocytic damage remains ambiguous. A possible role of altered expression of p21 in autistic-like behavior in rat offspring was examined with prenatal and postnatal BPA exposure. METHODS: Wistar albino dams were exposed to BPA (5 mg/kg) intraperitoneally throughout pregnancy and until the third postnatal day (PND). Pups were examined on 21st PND for behavioral test. Blood samples were collected for serum lactate levels and pups were sacrificed. Right frontal cortices were dissected out and processed for H&E, immunohistochemical analysis, and gene expression. RESULTS: Anxiety like behavior and thigmotaxis along with reduction in serum lactate concentrations were observed in pups exposed to BPA. Decline in neuronal number and decreased astrocytic population with reduced dendritic spines were revealed by H&E and immunohistochemical analysis, respectively, in right frontal cortices. Over expression of p21 was also detected in BPA-exposed offspring. CONCLUSIONS: Over expression of p21 may be associated with autistic behavior. Further studies are recommended to explore the structural alterations in other white matter pathways in frontal cortices.

Cr2O3-TiO2-Modified Filter Paper-Based Portable Nanosensors for Optical and Colorimetric Detection of Hydrogen Peroxide.

In the present approach, a Cr2O3-TiO2-modified, portable, and biomimetic nanosensor was designed to meet the requirement of a robust and colorimetric sensing of hydrogen peroxide. Cr2O3-TiO2 nanocomposites prepared via the hydrothermal method were fabricated as a transducer surface on the filter paper using the sol-gel matrix. The color on the filter paper sensor changed from green to blue upon the addition of hydrogen peroxide in the presence of TMB. This change in the color intensity was linear with the concentration of H2O2. RGB software was used as a color analyzing model to evaluate the optical signals. This paper-based colorimetric platform provided us with an improved analytical figure of merit with a linear range of 0.005-100 µM with 0.003 µM limit of detection. The real sample analysis and excellent anti-interference potential results proved the good analytical performance of the proposed design, providing a more promising tool for colorimetric H2O2 detection. Introducing Cr2O3-TiO2 nanocomposite-based paper sensors, being a novel method for optical and colorimetric detection, can pave the way for the development of other sensing devices for the detection of different analytes.

Epigenetic effects of in utero bisphenol A administration: Diabetogenic and atherogenic changes in mice offspring.

OBJECTIVES: Bisphenol A (BPA) that is a monomer of plastic products may possibly interfere with epigenetics and be involved in onset and progression of several diseases. This study was aimed to detect the epigenetic effects of in utero BPA exposure in mice offspring. MATERIALS AND METHODS: All experiments were performed according to the national guidelines for laboratory animals and after ethical approval. Thirty adult BALB/c female mice were divided into 3 equal groups, G1 (controls), G2 (ethanol 0.10 ml/100ml of PBS so that final concentration would be 0.01%) vehicle control and G3 (BPA 10 mg/kg). Chemicals were given twice a week throughout the pregnancy. Once delivered at term, female offspring were observed for body weight, behavior and movements. Blood glucose, serum insulin, cholesterol and high-density lipoprotein cholesterol (HDLc) were measured at 5 and 15 months postnatal. Animals were sacrificed at 15 months and pancreas, kidney, adipose tissue and uterine tissue were taken and stained with either Hematoxylin and eosin (H & E) or immunostaining and examined under light microscope. RESULTS: Offspring of group G3 revealed abnormal changes of body weight, behavior and movements. Blood glucose, serum insulin, cholesterol and HDLc were high in group G3 offspring compared to controls. H & E staining showed changes in the parenchyma of pancreas, kidneys and uterus, which were confirmed by staining with anti- islet-1, kidney-specific (Ksp) cadherin, and anti- MLH antibody. CONCLUSION: In utero exposure of BPA exerts diabetogenic and atherogenic effects with less parenchymal tissue in endocrine pancreas, kidney and uterus.

Antioxidant capacity and fatty acids characterization of heat treated cow and buffalo milk.

BACKGROUND: Antioxidant capacity of milk is largely due to vitamins A, E, carotenoids, zinc, selenium, superoxide dismutase, catalase, glutathione peroxidase and enzyme systems. Cow milk has antioxidant capacity while the antioxidant capacity of buffalo milk has been studied in a limited way. The information regarding the effect of pasteurization and boiling on antioxidant capacity of cow and buffalo milk is also scared. METHODS: Cow and buffalo milk was exposed to two different heat treatments i.e. 65 °C for 30 min and boiling for 1 min. After heat treatments, milk samples were cooled down to 4 °C packaged in transparent 250 ml polyethylene PET bottles and stored at 4 °C for 6 days. Milk composition, total flavonoid content, total antioxidant capacity, reducing power, DPPH free radical scavenging activity, antioxidant activity in linoleic acid, vitamin C, A, E, selenium, Zinc, fatty acid profile, peroxide value and sensory characteristics were studied in raw, pasteurized and boiled cow and buffalo milk at 0, 3 and 6 days of storage period. RESULTS: Total antioxidant capacity (TAC) of raw, pasteurized and boiled milk for cow (42.1, 41.3 and 40.7%) and buffalo (58.4, 57.6 and 56.5%) samples was found, respectively. Reducing power (RP) of raw cow and buffalo milk was 6.74 and 13.7 while pasteurization and boiling did not showed significant effect on RP of both cow and buffalo milk. DPPH activity of raw, pasteurized and boiled milk for cow (24.3, 23.8 and 23.6%) and buffalo (31.8, 31.5 and 30.4%) samples was noted, respectively. Storage period up to 3 days was non-significant while DPPH assay after 6 days of storage period indicated significant decline in antioxidant activity of milk samples. Antioxidant activity in linoleic acid (AALA) of buffalo and cow milk were recorded 11.7 and 17.4%, respectively. Pasteurization and boiling did not showed any impact on antioxidant capacity of cow and buffalo milk. The Loss of vitamin C in pasteurization (40 and 42%) and boiling (82 and 61%) of cow and buffalo milk was recorded, respectively. Concentration of vitamin A and E in pasteurized cow and buffalo milk was not significantly different from raw milk samples of cow and buffalo. Concentration of selenium and zinc was not influenced by the heat treatment in both cow and buffalo milk samples. After 3 days of refrigerated storage, antioxidant capacity of both cow and buffalo milk decreased. Concentrations of short-chain and medium-chain fatty acids increased in pasteurized and boiled cow and buffalo milk, while long-chain fatty acids decreased in pasteurized and boiled cow and buffalo milk, with no effect on colour and flavor score. Peroxide value of pasteurized and boiled cow and buffalo milk was not influenced by the storage up to 3 days. CONCLUSIONS: These results suggest that buffalo milk had a higher antioxidant capacity than cow milk and pasteurized milk should be consumed within 3 days of refrigerated storage for better antioxidant perspectives.

Pulmonary Alveolar Proteinosis: A Rare Cause of Respiratory Failure.

Pulmonary Alveolar Proteinosis (PAP) is a rare syndrome in the paediatric age group and characterized by intra-alveolar accumulation of proteinaceous phospholipid-laden material called surfactant. The diagnosis is made by High Resolution Computed Tomography (HRCT) chest which shows characteristic crazy paving appearance and diagnosis confirmed by Bronchoalveolar Lavage (BAL). We report two cases. First was a 9-month old infant who presented with respiratory distress and peripheral cyanosis since birth. He was diagnosed on High Resolution Computed Tomography (HRCT) chest as a case of pulmonary alveolar proteinosis and broncho-alveolar lavage confirmed his diagnosis. Second case was a 10-year old female child who had a history of repeated chest infections for 5 years and now presented with cough and respiratory distress for 45 days. She was also diagnosed on HRCT chest but unfortunately she died before bronchoalveolar lavage.

Effectiveness of nebulized N-acetylcysteine solution in children with acute bronchiolitis.

OBJECTIVE: To evaluate the effectiveness of nebulized mucolytic therapy (N-acetylcysteine) in treatment of children with acute bronchiolitis. STUDY DESIGN: Prospective randomized, controlled study. PLACE AND DURATION OF STUDY: Paediatric Medical Unit of the Children's Hospital and the Institute of Child Health, Lahore, from November 2011 to April 2012. METHODOLOGY: All the patients aged 2 months to 24 months, with the diagnosis of acute viral bronchiolitis were enrolled in the study. A predesigned proforma was completed after taking parental consent. All eligible patients were randomly assigned to one of the two groups: group-1 received N-acetylcysteine in nebulized form, and group-2 received salbutamol as a nebulized aerosol. Both groups were then compared regarding their clinical improvement and duration of stay in the hospital. The data obtained was analyzed on the statistical software SPSS version 16. Descriptive statistics were obtained by frequencies and percentages. Chi-square test was applied to find p-value. RESULTS: A total of 100 patients met the inclusion criteria. The mean age was 3 months. On the first day of treatment, the mean clinical severity score at baseline were 5.38 ± 2.62 in group-1 and 4.68 ± 2.2 in group-2. At day 3 and 5, it was 2.9 ± 1.48 and 3.30 ± 1.77 in group-1 and 0.88 ± 1.08 and 1.90 ± 1.32 in group-2 respectively. The clinical severity score after N-acetylcysteine inhalation therapy was better in group-1 on the third but more so on fifth day after hospital admission. The mean duration of hospitalization was 4.67 ± 2.2 days for the whole population and it differed between the 2 groups: 4.36 ± 1.66 days in group-1 versus 4.98 ± 2.6 days in group-2. CONCLUSION: On the basis of improvement in clinical severity score and early discharge from the hospital, N-acetylcysteine was found to be an effective therapy in acute bronchiolitis.

The novel heterozygous Thr377Arg MYOC mutation causes severe Juvenile Open Angle Glaucoma in a large Pakistani family.

Glaucoma is one of the major causes of blindness worldwide with characteristic optic disc changes and elevated intraocular pressure. It is subcategorized into Primary Open Angle Glaucoma (POAG) and Juvenile Open Angle Glaucoma (JOAG) depending upon age of the disease onset. Myocilin (MYOC) is the frequently mutated gene in familial cases of glaucoma. MYOC mutations show variable phenotype and penetrance. This study was aimed to identify disease causing mutation in 8 affected of a consanguineous family diagnosed with severe form of Juvenile Open Angle Glaucoma. Homozygosity mapping with four microsatellite markers and subsequent direct sequencing of MYOC revealed a novel heterozygous transition c.1130 C>G, substituting Threonine in to Arginine at codon 377 (p.Thr377Arg) of MYOC. This mutation was segregating with phenotype in all affected and was not found in control subjects. Ophthalmological findings revealed JOAG with severe and rapidly progressive phenotype. The age of onset was in the first decade of life and maximum Intra Ocular Pressure (IOP) recorded was 25mmHg. Bioinformatic tools predicted C to G transition at c.1130 as pathogenic and no structural changes were predicted in protein. This is the first report of novel MYOC mutation from Pakistan; segregating as autosomal dominant trait in large family diagnosed with JOAG. Identification of novel disease causing allele in MYOC indicates genetic heterogeneity of the population. This finding will help to provide genetic counseling to the affected family and carriers of this mutation may be advised for early therapeutic intervention to avoid irreversible visual loss.

Use of a pneumonia management tool to manage children with pneumonia at the first level health care facilities.

OBJECTIVE: To describe the application and evaluation of Pneumonia Management Tool (PMT) to manage children with non-severe pneumonia (NSP) at the first level health care (FLHC) facilities according to the standard case management (SCM) guidelines for acute respiratory infections (ARI). METHOD: The ARI SCM guidelines were simplified to a PMT and used by health workers at 14 FLHC facilities to assess, manage and monitor children with NSP and to educate caretakers on home care and follow-up visits. The district supervisors provided on the job support to various cadres of health workers of both public and private facilities. RESULTS: Of 949 children with NSP, 940 (99%) were successfully treated at FLHC facilities. Caretakers found PMT useful and of 1888 follow-up visits: 1872 (99.2%) brought PMT copy; 1627 (86.2%) brought their children to the facility; 1799 (95.3%) were on time and; 1857 (98.4%) had maintained antibiotic compliance. Using PMT, health workers adherence to SCM guidelines improved from 14% at baseline to 29% after training and 65% with on the job support. The practices remained similar among various cadres of health workers. CONCLUSIONS: Health workers used PMT in managing children with NSP, counselling caretakers on home care, follow-up visits and monitoring the treatment outcome. District level supervision helped to maintain a uniform skill enhancement.