Role of internalin proteins in the pathogenesis of Listeria monocytogenes.

Listeria monocytogenes is a food-borne bacterium that causes gastroenteritis, meningitis, or abortion. L. monocytogenes induces its internalization (entry) into human cells and either spreads laterally in tissues or transcytoses to traverse anatomical barriers. In this review, we discuss mechanisms by which five structurally related proteins of the "internalin" family of L. monocytogenes (InlA, InlB, InlC, InlF, and InlP) interact with distinct host receptors to promote infection of human cells and/or crossing of the intestinal, blood-brain, or placental barriers. We focus on recent results demonstrating that the internalin proteins InlA, InlB, and InlC exploit exocytic pathways to stimulate transcytosis, entry, or cell-to-cell spread, respectively. We also discuss evidence that InlA-mediated transcytosis contributes to traversal of the intestinal barrier, whereas InlF promotes entry into endothelial cells to breach the blood-brain barrier. InlB also facilitates the crossing of the blood-brain barrier, but does so by extending the longevity of infected monocytes that may subsequently act as a "Trojan horse" to transfer bacteria to the brain. InlA, InlB, and InlP each contribute to fetoplacental infection by targeting syncytiotrophoblast or cytotrophoblast layers of the placenta. This work highlights the diverse functions of internalins and the complex mechanisms by which these structurally related proteins contribute to disease.

Knockout of Dicer-2 in the Sf9 cell line enhances the replication of Spodoptera frugiperda rhabdovirus and conditionally increases baculovirus replication.

The Sf9 cell line, originally isolated from the ovarian tissue of Spodoptera frugiperda larvae, is widely used in academia and industry for the baculovirus-mediated production of recombinant proteins and virus-like particles. RNA interference (RNAi) is a conserved antiviral pathway present in eukaryotic organisms and is the primary antiviral defence mechanism in insects. Recent evidence has implicated RNAi as an antiviral response to baculovirus infection in Sf9 cells. To test this hypothesis, CRISPR/Cas9 technology was used to disable the RNAi pathway in Sf9 cells by knocking out Dicer-2, the protein responsible for cleaving viral double-stranded RNA precursors into short interfering RNAs. Infection of Dicer-2 knockout Sf9 cells with either the wild-type baculovirus Autographa californica nucleopolyhedrovirus (AcMNPV), recombinant AcMNPV (rAcMNPV) expressing ß-galactosidase (ß-gal), or rAcMNPV expressing a wasp venom protein (Vn50) at a multiplicity of infection (m.o.i.) of 1 resulted in a modest increase in virus replication compared to control Sf9 cells under adherent culture conditions. In contrast, Dicer-2 knockout Sf9 monolayer or suspension cultures infected by the rAcMNPV expressing ß-gal at higher m.o.i.s (3.5 and 20) did not exhibit increases in either viral DNA replication or ß-gal production. Intriguingly, during long-term passaging in suspension, Dicer-2 knockout Sf9 cultures underwent transient crashes in cell proliferation and viability. It was discovered that these periods of low growth and viability coincided with a dramatic increase in the RNA levels of S. frugiperda rhabdovirus, a recently identified adventitious virus that persistently infects the Sf9 cell line, suggesting a role for Dicer-2 in managing chronic viral infections in this industrially relevant insect cell line.

A Study on the Correlation between Change in the Geometrical Dimension of a Free-Falling Molten Glass Gob and Its Viscosity.

To produce flawless glass containers, continuous monitoring of the glass gob is required. It is essential to ensure production of molten glass gobs with the right shape, temperature, viscosity and weight. At present, manual monitoring is common practice in the glass container industry, which heavily depends on previous experience, operator knowledge and trial and error. This results in inconsistent measurements and consequently loss of production. In this article, a multi-camera based setup is used as a non-invasive real-time monitoring system. We have shown that under certain conditions, such as keeping the glass composition constant, it is possible to do in-line measurement of viscosity using sensor fusion to correlate the rate of geometrical change in the gob and its temperature. The correlation models presented in this article show that there is a strong correlation, i.e., 0.65, between our measurements and the projected viscosity.

Multi-Camera Based Setup for Geometrical Measurement of Free-Falling Molten Glass Gob.

High temperatures complicate the direct measurements needed for continuous characterization of the properties of molten materials such as glass. However, the assumption that geometrical changes when the molten material is in free-fall can be correlated with material characteristics such as viscosity opens the door to a highly accurate contactless method characterizing small dynamic changes. This paper proposes multi-camera setup to achieve accuracy close to the segmentation error associated with the resolution of the images. The experimental setup presented shows that the geometrical parameters can be characterized dynamically through the whole free-fall process at a frame rate of 600 frames per second. The results achieved show the proposed multi-camera setup is suitable for estimating the length of free-falling molten objects.

Antioxidant and Anti-Inflammatory Effects of Peganum harmala Extracts: An In Vitro and In Vivo Study.

Peganum harmala (P. harmala) belongs to the family Zygophyllaceae, and is utilized in the traditional medicinal systems of Pakistan, China, Morocco, Algeria, and Spain to treat several chronic health disorders. The aim of the present study was to identify the chemical constituents and to evaluate the antioxidant, anti-inflammatory, and toxicity effects of P. harmala extracts both in vitro and in vivo. Sequential crude extracts including 100% dichloromethane, 100% methanol, and 70% aqueous methanol were obtained and their antioxidant and anti-inflammatory effects evaluated both in vitro and in vivo. The anti-inflammatory effect of the extract was investigated using the carrageenan-induced paw edema method in mice, whereas the toxicity of the most active extract was evaluated using an acute and subacute toxicity rat model. In addition, we have used the bioassay-guided approach to obtain potent fractions, using solvent-solvent partitioning and reversed phase high performance liquid chromatography from active crude extracts; identification and quantification of compounds from the active fractions was achieved using electrospray ionization mass spectrometry and high performance liquid chromatography techniques. Results revealed that the 100% methanol extract of P. harmala exhibits significant in vitro antioxidant activity in DPPH assay with an IC50 of 49 µg/mL as compared to the standard quercetin with an IC50 of 25.4 µg/mL. The same extract exhibited 63.0% inhibition against serum albumin denaturation as compared to 97% inhibition by the standard diclofenac sodium in an in vitro anti-inflammatory assay, and in vivo anti-inflammatory against carrageenan-induced paw edema (75.14% inhibition) as compared to 86.1% inhibition caused by the standard indomethacin. Furthermore, this extract was not toxic during a 14 day trial of acute toxicity when given at a dose of 3 g/kg, indicating that the lethal dose (LD50) of P. harmala methanol extract was greater than 3 g/kg. P. harmala methanolic fraction 2 obtained using bioassay-guided fractionation showed the presence of quinic acid, peganine, harmol, harmaline, and harmine, confirmed by electrospray ionization mass spectrometry and quantified using external standards on high performance liquid chromatography. Taken all together, the current investigation further confirms the antioxidant, anti-inflammatory, and safety aspects of P. harmala, which justifies its use in folk medicine.

Effect of nebivolol beneficial on lipid profile and glycemic control in comparison with Atenolol in patients with type 2 DM with concomitant hypertension.

Blood pressure control in hypertensive patients with metabolic abnormalities is challenging because many antihypertensive drugs adversely affect metabolism. Nebivolol a 3rd generation vasodilatory ß-blocker offer neutral or beneficial effects on insulin sensitivity and lipid metabolism. The purpose of this study was to evaluate the effect of nebivolol and atenolol on glycemic control and lipid profile in type 2 diabetes patients with concomitant hypertension. We conducted a 12 weeks double blind randomized clinical trial at Sheik Zayed Hospital Rahim Yar Khan. Patients were randomly divided in to two groups. Patients in group were given tablet nebivolol 5-10mg while patients in group B were given tablet Atenolol 25-50mg/daily for a period of 12 weeks. Pre and post data were analyzed by SPSS 20. After 12 weeks, Both drugs lowered blood pressure significantly i.e. nebivolol (SBP from152±12 to130±14 with p=0.004, DBP from 95±12 to78±8.5 with p=0.002) Atenolol (SBP from148±16.5 to 128±15.5 with p=0.006, DBP from 90±10.5 to 82±12 with p=0.003).Similarly both Nebivolol and Atenolol did not any significant effect on glycemic control and lipid profile at 12 week with in groups. However when comparison was done between two groups, Nebivolol significantly reduced blood sugar (p=0.001), HbA1c (p=0.0032), total Cholesterol (p=0.002), triglycerides (p=0.012), LDL-Cholesterol (p=0.007) and HDL-Cholesterol (p=0.001) as compared to atenolol. In comparison with atenolol, Nebivolol has a beneficial effect on glycemic control and serum lipid profile.

The Host GTPase Arf1 and Its Effectors AP1 and PICK1 Stimulate Actin Polymerization and Exocytosis To Promote Entry of Listeria monocytogenes.

Listeria monocytogenes is a foodborne bacterium that causes gastroenteritis, meningitis, or abortion. Listeria induces its internalization (entry) into some human cells through interaction of the bacterial surface protein InlB with its host receptor, the Met tyrosine kinase. InlB and Met promote entry through stimulation of localized actin polymerization and exocytosis. How actin cytoskeletal changes and exocytosis are controlled during entry is not well understood. Here, we demonstrate important roles for the host GTPase Arf1 and its effectors AP1 and PICK1 in actin polymerization and exocytosis during InlB-dependent uptake. Depletion of Arf1 by RNA interference (RNAi) or inhibition of Arf1 activity using a dominant-negative allele impaired InlB-dependent internalization, indicating an important role for Arf1 in this process. InlB stimulated an increase in the GTP-bound form of Arf1, demonstrating that this bacterial protein activates Arf1. RNAi and immunolocalization studies indicated that Arf1 controls exocytosis and actin polymerization during entry by recruiting the effectors AP1 and PICK1 to the plasma membrane. In turn, AP1 and PICK1 promoted plasma membrane translocation of both Filamin A (FlnA) and Exo70, two host proteins previously found to mediate exocytosis during InlB-dependent internalization (M. Bhalla, H. Van Ngo, G. C. Gyanwali, and K. Ireton, Infect Immun 87:e00689-18, 2018, https://doi.org/10.1128/IAI.00689-18). PICK1 mediated recruitment of Exo70 but not FlnA. Collectively, these results indicate that Arf1, AP1, and PICK1 stimulate exocytosis by redistributing FlnA and Exo70 to the plasma membrane. We propose that Arf1, AP1, and PICK1 are key coordinators of actin polymerization and exocytosis during infection of host cells by Listeria.

Keys for Action: An Efficient Keyframe-Based Approach for 3D Action Recognition Using a Deep Neural Network.

In this paper, we propose a novel and efficient framework for 3D action recognition using a deep learning architecture. First, we develop a 3D normalized pose space that consists of only 3D normalized poses, which are generated by discarding translation and orientation information. From these poses, we extract joint features and employ them further in a Deep Neural Network (DNN) in order to learn the action model. The architecture of our DNN consists of two hidden layers with the sigmoid activation function and an output layer with the softmax function. Furthermore, we propose a keyframe extraction methodology through which, from a motion sequence of 3D frames, we efficiently extract the keyframes that contribute substantially to the performance of the action. In this way, we eliminate redundant frames and reduce the length of the motion. More precisely, we ultimately summarize the motion sequence, while preserving the original motion semantics. We only consider the remaining essential informative frames in the process of action recognition, and the proposed pipeline is sufficiently fast and robust as a result. Finally, we evaluate our proposed framework intensively on publicly available benchmark Motion Capture (MoCap) datasets, namely HDM05 and CMU. From our experiments, we reveal that our proposed scheme significantly outperforms other state-of-the-art approaches.

Effect of sitagliptin and glimepiride on C-reactive protein (CRP) in overweight Type-2 diabetic patients.

OBJECTIVES: To compare the anti-inflammatory effect of sitagliptin and glimepiride by measuring CRP in overweight Type-2 diabetic patients. METHODS: This clinical trial was conducted at diabetic clinic of Islam Central Hospital, Sialkot over a period of six months from June to November 2017. A total of 110 overweight Type-2 diabetic patients were divided in to two groups. Group-A was given tablet sitagliptin 50mg while Group-B was given tablet glimepiride 2mg for a period of 12 weeks. The dose was titrated according to blood sugar level. The primary outcome was measuring changes in CRP while secondary outcomes was changes in BMI, blood sugar, HbA1C, lipid profile and CRP from baseline in both study group using SPSS 16. RESULTS: After 12 weeks treatment, body weight increased in glimepiride but slightly reduced in sitagliptin, however comparison between them was non significant (p=0.07). Although both groups reduced blood sugar and HbA1c but comparison between them was non significant (p=0.59 and p=0.17 respectively) value. However lipid profile improved significantly in sitagliptin vs. glimepiride group i.e total cholesterol (-25±32.5 vs +1.5±45.4 P=0.02) triglycerides (-19±44.6 vs-1.8±48.7 P=0.001) LDL- cholesterol (-10±22.4 vs-0.8±18.7 P=0.001) HDL-cholesterol (-2.6±6.2 vs 1.2±5.2 P=0.03).Sitagliptin significantly reduced CRP in comparison to glimepiride (-2.3±1.8 vs0.8±1.5 P=0.001). CONCLUSION: Sitagliptin has strong anti inflammatory effect marked by reduction in CRP level in comparison to glimepiride in overweight type-2 diabetic patients. It also exerted beneficial effect on glycemic and lipid profiles.

Effect of vitamin D supplementation on various parameters in non-alcoholic fatty liver disease patients.

There are still no FDA approved drugs for NAFLD so far. Vitamin D may be a good therapeutic option for NAFLD patients due to its insulin sensitizing and anti-inflammatory properties. The purpose is to investigate the effect of oral vitamin D supplementation on various parameters in NAFLD patients. In this double blind randomized placebo controlled trial, 109 patients of NAFLD diagnosed by abdominal ultrasound and liver enzymes were divided into two groups for treatment with oral capsule of vitamin D3 50,000 IU and capsule placebo weekly for a period of 12 weeks. Anthropometric, chemical, metabolic and inflammatory parameters were assessed pre and post treatment by using SPSS 16. After 12 weeks oral treatment with vitamin D , its level increased significantly in vitamin D group from 12.5±4.2 to 24.5±3.8 ng/ml p =0.003 vs placebo group. This rise was further accompanied by decrease in HOMA-IR (4.56±1.6 to 3.26± 1.8 p=0.003) liver enzymes (i.e. ALT: 72.±17.6 to 54.5±14.5 IU/L p=0.04; AST: 68±14.5 to 46.± 10.5 p =0.002) serum CRP 3.25±0.68 to 2.28±0.44 mg/L p =0.06 and increase in serum adiponectin 8.56 ±1.12 to 10.44±2.35 mg/L p =0.03 as compared to placebo group. However non significant changes were observed in both groups in terms of body weight, BMI, and serum lipid profiles. Vitamin D supplementation not only improved its own status but also caused a significant amelioration in metabolic, chemical and inflammatory parameters in NAFLD patients. So it should be consider as an adjunctive therapy in NAFLD patients.

In vitro efficacy of linezolid against vancomycin resistant Enterococci.

Vancomycin-resistance among Enterococci has been escalating in recent years in many countries. Linezolid, an oxazolidinone is one of the novel drug that promised effective therapy of infections caused by vancomycin-resistant Enterococci (VRE).However, like with most of the previous antimicrobial agents, the clinical benefit of linezolid is being threatened by the emergence of resistant strains of MRSA and vancomycin resistant enterococci VRE, being reported in many countries. This study was conducted to establish linezolid susceptibility of VRE isolates by determining the in vitro activity of linezolid against vancomycin resistant Enterococci, isolated in our setup using fifty VRE isolates. The data collected from this study conclude that the vancomycin resistant Enterococci are 100% linezolid susceptible and presently there is no significant resistance against this unique oxazolidinone in our setup.

MicroRNA-like viral small RNA from Dengue virus 2 autoregulates its replication in mosquito cells.

MicroRNAs (miRNAs) are small regulatory RNAs that play significant roles in most cellular processes. In the seemingly endless arms race between hosts and pathogens, viruses also encode miRNAs that facilitate successful infection. In search of functional miRNAs or viral small RNAs (vsRNAs) encoded by Dengue virus (DENV), deep sequencing data of virus-infected Aedes aegypti mosquitoes were used. From six vsRNAs, with candidate stem-loop structures in the 5' and 3' untranslated regions of the viral genomic RNA, inhibition of DENV-vsRNA-5 led to significant increases in viral replication. Silencing of RNA interference (RNAi)/miRNA pathways' associated proteins showed that Argonaute 2 is mainly involved in DENV-vsRNA-5 biogenesis. Cloning of the precursor stem loop, immunoprecipitations, ectopic expression and detection in RNAi-deficient C6/36, and the mammalian Vero cell lines further confirmed DENV-vsRNA-5 production. Furthermore, significant impact of synthetic mimic and inhibitor of DENV-vsRNA-5 on DENV RNA levels revealed DENV-vsRNA-5's role in virus autoregulation by targeting the virus nonstructural protein 1 gene. Notably, DENV-vsRNA-5 homologous mimics from DENV serotypes 1 and 4, but not 3, inhibited DENV-2 replication. The results revealed that DENV is able to encode functional vsRNAs, and one of those, which resembles miRNAs, specifically targets a viral gene, opening an avenue for possible utilization of the small RNA to limit DENV replication.

Wolbachia small noncoding RNAs and their role in cross-kingdom communications.

In prokaryotes, small noncoding RNAs (snRNAs) of 50-500 nt are produced that are important in bacterial virulence and response to environmental stimuli. Here, we identified and characterized snRNAs from the endosymbiotic bacteria, Wolbachia, which are widespread in invertebrates and cause reproductive manipulations. Most importantly, some strains of Wolbachia inhibit replication of several vector-borne pathogens in insects. We demonstrate that two abundant snRNAs, WsnRNA-46 and WsnRNA-49, are expressed in Wolbachia from noncoding RNA transcripts that contain precursors with stem-loop structures. WsnRNAs were detected in Aedes aegypti mosquitoes infected with the wMelPop-CLA strain of Wolbachia and in Drosophila melanogaster and Drosophila simulans infected with wMelPop and wAu strains, respectively, indicating that the WsnRNAs are conserved across species and strains. In addition, we show that the WsnRNAs may potentially regulate host genes and Wolbachia genes. Our findings provide evidence for the production of functional snRNAs by Wolbachia that play roles in cross-kingdom communication between the endosymbiont and the host.

Pioglitazone attenuates cardiometabolic risk factors in non-diabetic patients with dyslipidemia.

OBJECTIVE: To determine the effect of pioglitazone on cardiometabolic risk factors in non-diabetic patients with dyslipidaemia. METHODS: This prospective, randomised clinical trial was conducted at Sheikh Zayed Medical College and Hospital, Rahim Yar Khan, Pakistan, from August to October 2016, and comprised non-diabetic patients with dyslipidaemia. They were randomly divided into three groups. First and second groups were given a daily dose of tab pioglitazone 30mg and gemfibrozil 600mg, respectively, while the third group served as the healthy control. Body weight, body mass index and serum lipid profile were analysed pre- and post-treatment. SPSS 16 was used for data analysis. RESULTS: Of the 135 participants, there were 45(33.3%) in each group. After 12 weeks' treatment, the pioglitazone group showed a highly significant reduction in body weight (83±10.5 to 76±13.5kg) and body mass index (27.7±4.4 to 25.5±6.4kg/m2) (p<0.01) compared to the gemfibrozil group. The pioglitazone group showed a significant improvement in serum lipid profile after 12 weeks (p<0.05) while the gemfibrozil group showed a highly significant improvement in serum lipid profile (p<0.01). CONCLUSIONS: Pioglitazone independently improved cardiometabolic risk factors, even in non-diabetics.

Effects of nigella sativa on various parameters in Patients of non-alcoholic fatty liver disease.

BACKGROUND: Non-alcoholic Fatty Liver disease (NAFLD) is the most common cause of progressive liver disorders worldwide. Drug options are limited with varying results. Nigella sativa in the form of herbal medicine could be another option because of its strong historical background. The objective of the study was to evaluate the effect Nigella sativa on various parameters in patients of NAFLD. METHODS: A randomized controlled trial was conducted at outpatient clinic of medical unit-1 of Sheikh Zayed Medical College/Hospital, Rahim Yar Khan, in which seventy patients of NAFLD were divided in to interventional and non-interventional groups. The interventional group was given cap Nigella sativa 1g twice a day while noninterventional group was given cap placebo in a same way for three months. Body weight, BMI, liver enzymes and ultrasound finding of fatty liver were assayed before and after treatment. RESULTS: After 12 weeks treatment with Nigella sativa body weight decreased significantly from 86±13.8 to76±12.6 kg vs placebo (p=0.041). BMI also reduced significantly from 29.06±4.6 to 26.25±6.2kg/m2 vs placebo (p=0.012). There is remarkable reduction in aminotransferases level after treatment with Nigella sativa vs placebo (ALT: 78.05±5.52 to 52.6±5.65 IU/L vs 76.48±4.95-74.32±5.58 IU/L (p=0.036). AST: 65.54±4.56-44.56±5.52 IU/L vs 63.25±5.43- 59.43±3.39 IU/L (p=0.021). There was overall 57.14 % patient had normal fatty liver grading on ultrasound after 12 weeks treatment with Nigella sativa as compared to placebo (p=0.002). CONCLUSIONS: Nigella sativa improves bio chemical and fatty liver changes in NAFLD patients. Its use in early stages of NAFLD is recommended in order to prevent its life-threatening complication.

Therapeutic benefits of green tea extract on various parameters in non-alcoholic fatty liver disease patients.

BACKGROUND AND OBJECTIVE: NAFLD affecting up to 30% of the population globally. Drug treatment options are limited with disappointing results. The dietary supplementation in the form of green tea is another option. Our objective was toinvestigate the effect of Green tea extract (GTE) supplementation on various parameters innon-alcoholicfatty liver disease (NAFLD) patients. METHODS: This study was conducted Dept. of Medicineof Sheikh Zayed Medical College/Hospital, Rahim Yar Khan from 15 April 2016 to 15 July 2016. Eighty overweight, non diabeticand dyslipidemic patients of NAFLD, diagnosed on the basis of ultrasound and aminotransferases level were randomized for treatmentwith capsule GTE500mg (n=40)and capsule placebo (n=40) twice a day for twelve weeks. Anthropometric parameters, liver enzymes, inflammatory markers and liver ultrasound imaging were estimated by SPSS-16 pre and post treatment. RESULTS: As compared to placebo, GTE caused a significant improvement in body weight (29.5±3.8 to 27.2±3.2 kg/m2 p=0.03), BMI (86±10.5 to 80±12.4 kg p=0.026), HOMA-IR(4.32±2.25 to 3.16± 1.6 p=0.0081) lipid profile (i.e. TC: L242.5±20.5 to 215.4±18.6 mg/dl p=0.005; TG: 175±22.6 to145±18 mg/dlp=0.003; LDL-C:155±12.5 to 140±16.7 mg/dl p=0.011; HDL-C: 36.8±6.7 to46.4±5.8 mg/dl p =0.001, Aminotransferases (i.e. ALT: 70.4±15.8to52.8±12.2 IU/L p=0.04; AST: 65.8±12.4 to 44.3± 8.5U/L p =0.002) and Inflammatory markers (hs-CRP: 3.14±0.58 to 2.18±0.32 p =0.023 Adiponectin: 8.46±1.02 to 10.55±3.42µg/ml p =0.003)GTE also caused a 67.5% regression of fatty liver changes on ultrasound as compared to placebo which is 25%only. CONCLUSION: GTEtherapy resulted in significant improvement in metabolic, chemical, inflammatory and radiological parameters of non-alcoholic fatty liver disease patients who were non-diabetic anddyslipidemic.

Vitamin D supplementation improves FEV1 in patients of Bronchial Asthma.

BACKGROUND AND OBJECTIVE: Vitamin D deficiency has strong association with various respiratory disorders in which bronchial asthma is one of them. The objective was to determine the efficacy of vitamin D supplementation in cases with bronchial asthma. METHODS: This case control study was conducted at private clinical set up of district, Rahim Yar Khan from August to October 2016 in which 100 cases of bronchial asthma were randomly divided into Group-A and Group-B each contained 50 patients. Group-A was given placebo and Group-B with vitamin D in a dose of 50,000 units per day orally. Both the groups were followed in terms of improvement in FEV1 at 1, 2 and 3 months. RESULTS: There was no significant difference in both groups in terms of BMI and duration of asthma at start of study. The mean pre treatment vitamin D level of Group-A was 14.23±1.66 and of Group-B, 15.30±2.05 ng/dl (p= 0.23). FEV1 in pre treatment Group-A was 64.35±3.16 and of Group-B was 62.35±2.16 with p= 0.95. There was no significant difference in terms of FEV1 in both the groups at one month (p= 0.32). While at two months it was significantly higher in Group-B with p= 0.04. At 3 months the final outcome was seen where the post treatment FEV1 in Group-A was 66.13±2.75 and in Group-B, 75.15±2.04 with p value of 0.001. CONCLUSION: Vitamin D supplementation improves FEV1 significantly at two months and these can be even highly significant if it is extended up to 3 months.

Neutrophil lymphocyte ratio (NLR): A well assessment tool of glycemic control in type 2 diabetic patients.

BACKGROUND AND OBJECTIVE: Increased neutrophil lymphocyte ratio (NLR) is a marker as well as predictor of various cardiac and non cardiac disorders. Our aim was to assess the relationship between NLR and different level of glycemic control in type 2 diabetic patients. METHODS: An observational study was conducted at diabetic clinic of Sheikh Zayed Medical College/Hospital, Rahim Yar Khan from September 2016 to February 2017 in which 330 type 2 diabetic patients were randomly divided in to three groups based upon diabetes control according to ADA criteria. Patients in group A with HbA1c ≤ 7% (excellent control), group B HbA1c 7.0-9.0 % (poor control) and group C HbA1c ≥ 9 %(worst control). Patients were assessed in terms of complete blood count and C - reactive protein. RESULTS: As compared to excellent control (Group A) patients with worst control (Group C)showed a high leukocyte count (p.001), high neutrophil count (P.003) and lower lymphocyte count (P 0.44) while patients in poor control (Group B)did not differ significantly. Similarly value of NLR was also significantly higher in worst control (Group C) as compared to poor control(Group B) and excellent control (Group A) diabetes (4.3±2.8, 2.7±1.0 and2.0±0.5(p.001). NLR were found independent predictor of worst diabetes control (OR: 1.809, 95% CI: 1.459-2.401) along with fasting blood sugar (OR: 0.938, 95% CI: 0.995-0.982) and CRP (OR: 1.020, 95% CI: 1.003-1.028). CONCLUSION: Increased NLR level is associated with elevated HbA1c and poor glycemic control in patients of type 2 diabetes mellitus. It can be used as a disease monitoring tool during the follow up of diabetic patients.

The Baculovirus Antiapoptotic p35 Protein Functions as an Inhibitor of the Host RNA Interference Antiviral Response.

UNLABELLED: RNA interference (RNAi) is considered an ancient antiviral defense in diverse organisms, including insects. Virus infections generate double-strand RNAs (dsRNAs) that trigger the RNAi machinery to process dsRNAs into virus-derived short interfering RNAs (vsiRNAs), which target virus genomes, mRNAs, or replication intermediates. Viruses, in turn, have evolved viral suppressors of RNAi (VSRs) to counter host antiviral RNAi. Following recent discoveries that insects mount an RNAi response against DNA viruses, in this study, we found that Autographa californica multiple nucleopolyhedrovirus (AcMNPV) infection similarly induces an RNAi response in Spodoptera frugiperda cells by generating a large number of vsiRNAs postinfection. Interestingly, we found that AcMNPV expresses a potent VSR to counter RNAi. The viral p35 gene, which is well known as an inhibitor of apoptosis, was found to be responsible for the suppression of RNAi in diverse insect and mammalian cells. The VSR activity of p35 was further confirmed by a p35-null AcMNPV that did not suppress the response. In addition, our results showed that the VSR activity is not due to inhibition of dsRNA cleavage by Dicer-2 but acts downstream in the RNAi pathway. Furthermore, we found that the VSR activity is not linked to the antiapoptotic activity of the protein. Overall, our results provide evidence for the existence of VSR activity in a double-stranded DNA virus and identify the responsible gene, which is involved in the inhibition of RNAi as well as apoptosis. IMPORTANCE: Our findings demonstrate the occurrence of an insect RNAi response against a baculovirus (AcMNPV) that is highly utilized in microbial control, biological and biomedical research, and protein expression. Moreover, our investigations led to the identification of a viral suppressor of RNAi activity and the gene responsible for the activity. Notably, this gene is also a potent inhibitor of apoptosis. The outcomes signify the dual role of a virus-encoded protein in nullifying two key antiviral responses, apoptosis and RNAi.

Detection of Carbofuran-Protein Adducts in Serum of Occupationally Exposed Pesticide Factory Workers in Pakistan.

This study was conducted to investigate the protein adducts with pesticides in a cohort of 172 factory workers that were exposed to a mixture of pesticides. The 35 samples showing considerable variation in biochemical parameters, i.e., butyrylcholinestrase (BChE), serum glutamic pyruvic transaminase (SGPT), serum glutamic oxaloacetic transaminase (SGOT), gamma-glutamyl transferase (GGT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase (ALP/ALKP), lactate dehydrogenase (LDH), creatine phosphokinase (CPK) enzymes, and controls were analyzed by reversed-phase nanoscale liquid chromatography tandem mass spectrometry (nLC-MS/MS) on an Orbitrap mass spectrometer employing a shotgun proteomics approach. Only protein adducts with carbofuran were found on serum proteins of these workers. These adducts were of carbofuran labeled lysine (Lys-142, Lys-183, Lys-287, and Lys-467), arginine (Arg-210, Arg-242, and Arg-256) from serum albumin, and serine (Ser-07, Ser-54, and Ser-150) from immunoglobulin proteins. The arginine residues (Arg-210, Arg-242, Arg-246, and Arg-434) from albumin were also found to be glycated in serum of workers showing a high level of glucose who also had glycated arginine (Arg-1120) modified with carbofuran in their tankyrase-1-binding protein. The number of tandem mass spectra of modified peptides increased with increasing time of exposure. This is the first report to demonstrate the presence of carbofuran-labeled albumin, immunoglobulin, and glycated arginine, which shows that lysine and arginine of human albumin and serine of immunoglobulin are covalently modified in the serum of workers that were occupationally exposed to carbofuran, and the modification is detectable by tandem mass spectrometry. These peptides modified with carbofuran can potentially be used as a biomarker of carbofuran exposure.