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The purpose of this study is to assess the effects of early high-carbohydrate stimulus on glucose metabolism in zebrafish (Danio rerio) over two generations and explore the mechanisms that explain those nutritional programming effects via epigenetic modifications. The larvae were delivered a high-carbohydrate diet (53.66%) that was used as an early nutritional stimulus from the first feeding to the end of the yolk sac (FF) and 5 days after yolk-sac exhaustion (YE). The larvae (F0) and their offspring (F1) were then both fed the control diet (22.69%) until adulthood (15 weeks), and they were challenged with a high-carbohydrate diet (35.36%) at the 16th week. The results indicated that early stimulus immediately raised the mRNA levels of genes involved in glycolysis and gluconeogenesis. At the end of F0 challenge, both treatment groups decreased the plasma glucose levels, increased the expression levels of glucokinase (gck), and inhibited the mRNA during gluconeogenesis. When challenged in F1, the glucose levels were lower in FF (F1), and the mRNA levels of phosphoenolpyruvate carboxykinase 1 (pck1) were decreased in FF (F1) and YE (F1). Besides, in both experimental groups (F0 and F1), the CpG island of pck1 maintained lower levels of hypermethylated expression from F0 adult, 24 h post-fertilization embryo, to F1 adult. In conclusion, these results indicated that an early high-carbohydrate stimulus could significantly reprogram glucose metabolism in adult zebrafish, that those modifications could be partially transmitted to the next generation, and that the DNA methylation of pck1 might work as a stable epigenetic marker to contribute to those processes.

Malnutrition is one of the major causes in low and middle-income families’ children’s deaths under the age of 5 years. Nine times as many children with severe acute malnutrition (SAM) will pass away than healthy youngsters. In Pakistan, 17.7% of children under five are wasted, and 4 out of 10 infants under five are stunted. With almost 1/3 children (28.9%) underweight, the double burden of malnutrition is increasing. Objectives: To assess and identify the factors influencing malnutrition among under five years children of district Gwadar. Methods: A community-based cross-sectional research was carried out in randomly chosen clusters in several union councils in the Pakistani district of Gwadar. Mothers and other caregivers of children under five were interviewed as part of the house-to-house survey, using a practical sample approach. Results: The children age mean was 11.56 ± 12.4 months and the mean age of mothers was 27.29 ± 6.31 years. Overall stunting prevalence of severe acute malnutrition (SAM) cases was 21.2% and moderate acute malnutrition (MAM) cases were 46.2%. Conclusions: According to the study's results, under the age of five the malnutrition was widespread. The three types of nutritional outcomes were all strongly correlated with household income. Particularly in Baluchistan's underdeveloped regions, more labor and studies are needed

Introduction: It is vital to diagnose pneumothorax at early stage for treatment purpose. In trauma cases usually it is ignored and remained undiagnosed which can become cause of cardiac arrest and tension. Ultrasound has helped the trauma cases to diagnose the pneumothorax by performing chest ultrasound and can manage the treatment plan and resuscitation. Objective: The purpose of the study is to analyze the diagnostic accuracy of chest ultrasound in identifying the pneumothorax from trauma cases while CT remain as a gold standard. Methodology: The trauma patients who were referred to Radiology Department of Lahore General Hospital from the period of January 2019 to January 2020 were included in the study. Total 152 patients were included in the study. The ultrasound was performed in supine position by using high frequency linear probe by scanning and focusing on both lungs and identified the parietal and visceral pleura, comet tail artifact and lung sliding which shows normal lungs. The gold standard set was CT results of pneumothorax. Results: Total of 152 patients who have chest trauma were included and the mean age of the selected sample was 31.6 ± 13.4 years (range: 5-68).Male patients were more in number (76%).Pneumothorax was found in 55 cases which counts 36.2% of the total cases. The sensitivity of the ultrasound was 83.6 %. Specificity of the ultrasound was 97.9%. Positive predictive value of ultrasound was 95.8% and negative predictive value 91.3%. Conclusion: Chest ultrasound is accurate, no harm of radiation and quick technique to identify the pneumothorax in chest trauma patients in medical emergency. Key Words: Chest Trauma, Pneumothorax, CT scan, Ultrasonography

Introduction: Antenatal care needs frequent examination to analyze the fetal growth. As the time progress the weight of the neonate increases. Measuring weight for identifying the fetal growth pattern in three terms of the pregnancy is therefore very important especially in high risk pregnancies. Ultrasound helps to measure the fetal weight in order to estimate the birth weight. Estimated birth weight helps to plan the delivery options and is a useful parameter for fetal growth. Objective: The aim of the study is to identify the difference between the actual birth weight of neonates and the estimated birth weight calculated by the ultrasound scan in primipara and multipara. Method: The present study was conducted in Lahore General Hospital Radiology Department from June 20th 2019 to July 10th 2019. Pregnant women were included in the study that was falling in their last trimester of pregnancy especially after 37th week of pregnancy and above. Fetus weight from the ultrasound estimation was recorded and the actual birth weight also recorded at the time of delivery to make actual comparison. Sample size of the study was 282 pregnant women. Results: Advancement in medical sciences has helped to reduce the mortality rate among pregnant women and in neonates. The estimation of birth weight with the help of ultrasound has helped the women to diagnose any medical and growth issue before time and from the study there was found no significant gap between the estimated birth weight and actual birth weight. Conclusion: Ultrasound is noninvasive and safe tool to estimate and assess the fetal birth weight for better decision about mode of delivery

The current study aimed at azo dye degradation using a facultative anaerobe. A bacterial strain capable of decolorization and degradation of RB5 under static conditions was isolated from root nodules of Cactus. The strain efficiently degraded Reactive Black 5 (RB5) with metabolites detected by GC–MS. The strain was identified as Proteus mirabilis and placed in phylogenetic position through 16S rRNA gene analysis. The degradation increased with time to achieve up to 90 % after 72 h incubation at 37 oC and pH value of 7. Variations of C source did not affect the process while yeast extract was effective. The effective degradation was achieved at 100 ppm of dye. The GC–MS detected metabolites included; acetaldoxime, oxirane trimethyl, acetic acid-2-propenyl ester, acetaldehyde semi carbazone, 2-pentanone, 4-hydroxy-4-methyl, hydrazine, 1-5-hexenyl-1-methyl and enanthamide Protox and phytotoxicity results showed fewer toxic ranges of these metabolites as compared to products of benzene and amines. The isolated strain can be useful in the bioremediation of azo dyes contaminated environments.

There was an error published in J. Cell Sci. (2013) 126, [3649–3663][1] ([doi:10.1242/jcs.125088][2]). The affiliations for Marwa Eltoweissy were incorrect. The correct affiliations are as given below. Hassan Dihazi1, Gry Helene Dihazi1, Asima Bibi1, Marwa Eltoweissy1,2, Claudia A. Mueller3

Due to the exponential increase in the human population of this bio-sphere, energy resources are becoming scarce. Because of the traditional methods, most of the generated energy is wasted every year in the distribution network and demand side. Therefore, researchers all over the world have taken a keen interest in this issue and finally introduced the concept of the smart grid. Smart grid is an ultimate solution to all of the energy related problems of today&rsquo, s modern world. In this paper, we have proposed a meta-heuristic optimization technique called the dragonfly algorithm (DA). The proposed algorithm is to a real-world problem of single and multiple smart homes. In our system model, two classes of appliances are considered, Shiftable appliances and Non-shiftable appliances. Shiftable appliances play a significant role in demand side load management because they can be scheduled according to real time pricing (RTP) signal from utility, while non-shiftable appliances are not much important in load management, as these appliances are fixed and cannot be scheduled according to RTP. On behalf of our simulation results, it can be concluded that our proposed algorithm DA has achieved minimum electricity cost with a tolerable waiting time. There is a trade-off between electricity cost and waiting time because, with a decrease in electricity cost, waiting time increases and vice versa. This trade-off is also obtained by our proposed algorithm DA. The stability of the grid is also maintained by our proposed algorithm DA because stability of the grid depends on peak-to-average ratio (PAR), while PAR is reduced by DA in comparison with an unscheduled case.

The thick ascending limb of Henle's loop (TALH) is normally exposed to variable and often very high osmotic stress and involves different mechanisms to counteract this stress. ER resident calcium binding proteins especially calreticulin (CALR) play an important role in different stress balance mechanisms. To investigate the role of CALR in renal epithelial cells adaptation and survival under osmotic stress, two-dimensional fluorescence difference gel electrophoresis combined with mass spectrometry and functional proteomics were performed. CALR expression was significantly altered in TALH cells exposed to osmotic stress, whereas renal inner medullary collecting duct cells and interstitial cells exposed to hyperosmotic stress showed no significant changes in CALR expression. Moreover, a time dependent downregulation of CALR was accompanied with continuous change in the level of free intracellular calcium. Inhibition of the calcium release, through IP3R antagonist, prevented CALR expression alteration under hyperosmotic stress, whereas the cell viability was significantly impaired. Overexpression of wild type CALR in TALH cells resulted in significant decrease in cell viability under hyperosmotic stress. In contrast, the hyperosmotic stress did not have any effect on cells overexpressing the CALR mutant, lacking the calcium-binding domain. Silencing CALR with siRNA significantly improved the cell survival under osmotic stress conditions. Taken together, our data clearly highlight the crucial role of CALR and its calcium-binding role in TALH adaptation and survival under osmotic stress.

Renal fibrosis is characterized by excessive accumulation of extracellular matrix (ECM), which compromises organ function by replacing normal organ tissue. The molecular mechanisms leading to renal fibrosis are not fully understood. Here we demonstrated that TGFβ1, AGT or PDGF stimulation of renal cells resulted in endoplasmic reticulum (ER) stress followed by activation of the protective unfolded protein response pathway and a high secretory level of protein disulfide isomerase ERP57 (also known as PDIA3). The TGFβ1-induced impairment of ER function could be reversed by treatment with BMP7, suggesting a specific involvement in renal fibrosis. A clear correlation between the degree of fibrosis, ER stress and the level of ERP57 could be seen in fibrosis animal models and in biopsies of renal fibrosis patients. Protein interaction studies revealed that secreted ERP57 exhibits a strong interaction with ECM proteins. Knockdown of ERP57 or antibody-targeted inhibition of the secreted form significantly impaired the secretion and accumulation of ECM. Moreover, ERP57 was excreted in the early stages of chronic kidney disease, and its level in urine correlated with the degree of renal fibrosis, suggesting that the secretion of ERP57 represents one of the first signs of renal fibrosis onset and progression.

AIM: To investigate the proteome changes of stem cells due to ciclopirox olamine (CPX) treatment compared to control and retinoic acid treated cells. METHODS: Stem cells (SCs) are cells, which have the ability to continuously divide and differentiate into various other kinds of cells. Murine embryonic stem cells (ESCs) and multipotent adult germline stem cells (maGSCs) were treated with CPX, which has been shown to have an antiproliferative effect on stem cells, and compared to stem cells treated with retinoic acid (RA), which is known to have a differentiating effect on stem cells. Classical proteomic techniques like 2-D gel electrophoresis and differential in-gel electrophoresis (DIGE) were used to generate 2D protein maps from stem cells treated with RA or CPX as well as from non-treated stem cells. The resulting 2D gels were scanned and the digitalized images were collated with the help of Delta 2D software. The differentially expressed proteins were analyzed by a MALDI-TOF-TOF mass spectrometer, and the identified proteins were investigated and categorized using bioinformatics. RESULTS: Treatment of stem cells with CPX, a synthetic antifungal clinically used to treat superficial mycoses, resulted in an antiproliferative effect in vitro, without impairment of pluripotency. To understand the mechanisms induced by CPX treatments which results in arrest of cell cycle without any marked effect on pluripotency, a comparative proteomics study was conducted. The obtained data revealed that the CPX impact on cell proliferation was accompanied with a significant alteration in stem cell proteome. By peptide mass fingerprinting and tandem mass spectrometry combined with searches of protein sequence databases, a set of 316 proteins was identified, corresponding to a library of 125 non-redundant proteins. With proteomic analysis of ESCs and maGSCs treated with CPX and RA, we could identify more than 90 single proteins, which were differently expressed in both cell lines. We could highlight, that CPX treatment of stem cells, with subsequent proliferation inhibition, resulted in an alteration of the expression of 56 proteins compared to non-treated cells, and 54 proteins compared to RA treated cells. Bioinformatics analysis of the regulated proteins demonstrated their involvement in various biological processes. To our interest, a number of proteins have potential roles in the regulation of cell proliferation either directly or indirectly. Furthermore the classification of the altered polypeptides according to their main known/postulated functions revealed that the majority of these proteins are involved in molecular functions like nucleotide binding and metal ion binding, and biological processes like nucleotide biosynthetic processes, gene expression, embryonic development, regulation of transcription, cell cycle processes, RNA and mRNA processing. Proteins, which are involved in nucleotide biosynthetic process and proteolysis, were downregulated in CPX treated cells compared to control, as well as in RA treated cells, which may explain the cell cycle arrest. Moreover, proteins which were involved in cell death, positive regulation of biosynthetic process, response to organic substance, glycolysis, anti-apoptosis, and phosphorylation were downregulated in RA treated cells compared to control and CPX treated cells. CONCLUSION: The CPX treatment of SCs results in downregulation of nucleotide binding proteins and leads to cell cycle stop without impairment of pluripotency. peerReviewed

Renal fibrosis is characterized by excessive accumulation of extracellular matrix (ECM), which compromises organ function by replacing normal organ tissue. The molecular mechanisms leading to renal fibrosis are not fully understood. Here we demonstrated that TGFβ1, AGT or PDGF stimulation of renal cells resulted in endoplasmic reticulum (ER) stress followed by activation of the protective unfolded protein response pathway and a high secretory level of protein disulfide isomerase ERP57 (also known as PDIA3). The TGFβ1-induced impairment of ER function could be reversed by treatment with BMP7, suggesting a specific involvement in renal fibrosis. A clear correlation between the degree of fibrosis, ER stress and the level of ERP57 could be seen in fibrosis animal models and in biopsies of renal fibrosis patients. Protein interaction studies revealed that secreted ERP57 exhibits a strong interaction with ECM proteins. Knockdown of ERP57 or antibody-targeted inhibition of the secreted form significantly impaired the secretion and accumulation of ECM. Moreover, ERP57 was excreted in the early stages of chronic kidney disease, and its level in urine correlated with the degree of renal fibrosis, suggesting that the secretion of ERP57 represents one of the first signs of renal fibrosis onset and progression.

Renal fibrosis is a process that is characterized by declining excretory renal function. The molecular mechanisms of fibrosis are not fully understood. Oxidative stress pathways were reported to be involved in renal tissue deterioration and fibrosis progression. In order to identify new molecular targets associated with oxidative stress and renal fibrosis, differential proteomics analysis was performed with established renal cell lines (TK173 and HK-2). The cells were treated with oxidative stress triggering factor H(2)O(2) and the proteome alterations were investigated. Two dimensional protein maps were generated and differentially expressed proteins were processed and identified using mass spectrometry analysis combined with data base search. Interestingly the increase of ROS in the renal cell lines upon H(2)O(2) treatment was accompanied by alteration of a large number of proteins, which could be classified in three categories: the first category grouped the proteins that have been described to be involved in fibrogenesis (e.g. ACTA2, VIN, VIM, DES, KRT, COL1A1, COL4A1), the second category, which was more interesting involved proteins of the oxidative stress pathway (PRDX1, PRDX2, PRDX6, SOD, PARK7, HYOU1), which were highly up-regulated under oxidative stress, and the third category represented proteins, which are involved in different other metabolic pathways. Among the oxidative stress proteins the up-regulation of PARK7 was accompanied by a shift in the pI as a result of oxidation. Knockdown of PARK7 using siRNA led to significant reduction in renal cell viability under oxidative stress. Under H(2)O(2) treatment the PARK7 knockdown cells showed up to 80% decrease in cell viability and an increase in apoptosis compared to the controls. These results highlight for the first time the important role of PARK7 in oxidative stress resistance in renal cells. peerReviewed

Renal fibroblasts are thought to play a major role in the development of renal fibrosis (RF). The mechanisms leading to this renal alteration remain poorly understood. We performed differential proteomic analyses with two established fibroblast cell lines with RF phenotype to identify new molecular pathways associated with RF. Differential 2-DE combined with mass spectrometry analysis revealed the alteration of more than 30 proteins in fibrotic kidney fibroblasts (TK188) compared to normal kidney fibroblast (TK173). Among these proteins, markers of the endoplasmic reticulum (ER) stress- and the unfolded protein response (UPR) pathway (GRP78, GRP94, ERP57, ERP72, and CALR) and the oxidative stress pathway proteins (PRDX1, PRDX2, PRDX6, HSP70, HYOU1) were highly up-regulated in fibrotic cells. Activation of these stress pathways through long time exposition of TK173, to high NaCl or glucose concentrations resulted in TK188 like phenotype. Parallel to an increase in reactive oxygen species, the stressed cells showed significant alteration of fibrosis markers, ER-stress and oxidative stress proteins. Similar effects of osmotic stress could be also observed on renal proximal tubule cells. Our data suggest an important role of the ER-stress proteins in fibrosis and highlights the pro-fibrotic effect of osmotic stress through activation of oxidative stress and ER-stress pathways.