EIA Kit Research Articles

Interlude of cGMP and cGMP/protein kinase G type 1 in pancreatic adenocarcinoma cells.

cAMP and cGMP signaling is important both for normal and cancer cells. This signaling is controlled by adenylyl and guanylyl cyclases and cyclic nucleotide phosphodiesterases. One of the direct targets for cGMP is protein kinase G (PKG). The main aim of this work was to investigate cGMP and PKG signaling in pancreatic adenocarcinoma (PDAC) cells. The PKG activity, cGMP, and calcium level were measured with the CycLex Cyclic GMP dependent protein kinase (cGK) Assay Kit, the DetectX Cyclic GMP Colorimetric EIA Kit, and the Fluo-4 NW Calcium Assay Kit, respectively. The Proteome Profiler Array was done using Human Phospho-Kinase Array and Human Phospho-MAPK Array Kits. This study shows for the first time that functional PKG1 is expressed in PDAC cells. It demonstrates that the specific PKG1 inhibitor, DT3, induces cytotoxicity through necrosis and reduces proliferation and migration of PDAC cells. Moreover, ERK1/2 and p38 can be considered as potential targets for PKG1 in PDAC cells. In addition, the study shows that phosphodiesterases and nitric oxide-guanylyl cyclases regulate the cGMP level in PDAC cells, affecting the proliferation of the cells. The cGMP and PKG signaling may be a target for developing new therapeutic approaches for PDAC.

Complete feminization of catfish by feeding Limnodilus, an annelid worm collected in contaminated streams

BackgroundFeminization of animals derived from areas polluted by endocrine disrupting chemicals (EDCs) has been observed in all classes of vertebrates. However, feminization of artificially reared offspring by feeding of specific living organisms has never been reported. MethodsDifferent food (including Limnodilus spp collected from the wild) and time treatment were applied to southern catfish. In addition, EDCs in Limnodilus spp., an annelid worm collected from wild contaminated small streams, was detected by LC–MS (Liquid chromatography-mass spectrometry). Serum estradiol-17β and vitellogenin (VTG) levels and gonadal Sf1, Dmrt1, Foxl2, Cyp19a1a expression levels in the catfish were measured through Estradiol/VTG EIA Kit and real-time PCR. ResultsHere we report that feeding of Limnodilus spp. resulted in complete feminization of southern catfish, which has a 1:1 sex ratio in wild conditions. Furthermore, HPLC analysis showed that the extraction of Limnodilus spp. contained EDCs, including bisphenol A (BPA), diethylstilbestrol (DES), 4-tert-octylphenol (4-t-OP) and 4-nonylphenol (4-NP), which were further confirmed by LC–MS. Feeding southern catfish using commercial diets sprayed with EDCs cocktail also resulted in 100% female, whereas the control fish displayed approximate 1:1 sex ratio. Limnodilus spp. fed fish displayed similar serum estradiol-17β and VTG levels and gonadal Sf1, Dmrt1, Foxl2, Cyp19a1a expression levels to those of female control. ConclusionThese results demonstrated that EDCs in Limnodilus spp. cause southern catfish feminization by affecting aromatase expression and endogenous estrogen level. This is the first report showing that feeding of any living organism resulted in complete feminization of a vertebrate.

Significant correlation between salivary and serum Ca 15-3 in healthy women and breast cancer patients.

The tumor marker CA 15-3 is one of the most import reliable for metastatic breast cancer monitoring. While it is generally assessed in serum of patients, blood sampling is an invasive method compared to saliva sampling which is simple and could be an alternative to blood according to many studies. The aim of this investigation was to assess the relationship between serum and salivary concentrations of the protein CA 15-3 in patients with breast cancer and healthy asymptomatic volunteers. A case-control study was conducted with 60 women: 29 breast cancer patients from the Maternity Hospital Souissi Rabat (Morocco) and 31 healthy asymptomatic women. The CA 15-3 concentrations in saliva and serum samples were assessed using an enzyme immune assay (EIA kits) and comparison between cases and controls was made by the Mann-Whitney test. The correlation between serum and saliva CA 15-3 concentration was tested using Pearson correlation. The comparison result of CA15-3 concentration in saliva and serum level in cases and controls was not statistically significant (p>0.05). However, the correlation between salivary and serum CA 15-3 concentration was positive and statistically significant (r=0.27, p=0.03). In conclusion, the positive correlation between salivary and serum expression found in our study suggests that saliva could be an alternative to blood sampling to help breast cancer monitoring.

SAT0440 Serum Sclerostin is Higher in Men with Severe Osteophytes at the Spine – the Minos Study

BackgroundSclerostin is a negative regulator of bone formation that might be involved in osteoarthritis (OA) pathophysiology.ObjectivesOur aim was to analyze the association between spine OA severity and serum sclerostin concentration...

Alditols and monosaccharides from sorghum vinegar can attenuate platelet aggregation by inhibiting cyclooxygenase-1 and thromboxane-A2 synthase

Ethnopharmacological relevanceVinegar has been used as both a common seasoning and a traditional Chinese medicine. Sorghum vinegar is an excellent source of physiological substances with multiple health benefits. Aim of this studyTo evaluate the antiplatelet aggregation activity of alditols and monosaccharides extracted from sorghum vinegar and analysis its mechanism. Materials and methodsAlditol and monosaccharide extract (AME) from sorghum vinegar was first evaluated for antiplatelet activity using the turbidimetric method. Blood was collected from healthy volunteer donors. The platelet aggregation was induced by arachidonic acid (AA), collagen, adenosine diphosphate (ADP) and thrombin in vitro. AME was divided into three experimental groups with the concentration were 0.10, 0.25 and 0.50mg/mL. In order to determine the inhibitory activity of AME on COX1, TXS and TXA2 production experiments were conducted using the COX1, TXS and TXB2 EIA kit. Computational docking was used to find the docking pose of monosaccharides and alditols with COX1. ResultsAME showed significant induction of antiplatelet activity by arachidonic acid (AA), collagen, adenosine diphosphate (ADP) and thrombin in a concentration-dependent manner (p<0.05). AME (0.50mg/mL) reduced the AA-induced aggregation rate to 10.35%±0.46%, which was comparable to acetylsalicylic acid (aspirin, ASA) (0.50mg/mL, 6.35%±0.58%), a medical standard. Furthermore, AME strongly inhibited cyclooxygenase-1 (COX1) and thromboxane-A2 synthase (TXS), and subsequently attenuated thromboxane-A2 (TXA2) production. These findings indicated that AME attenuates platelet aggregation through the AA metabolism pathway. Computational docking showed that alditols (L-erythritol, L-arabitol, xylitol and D-sorbitol), monosaccharides (D-glucopyranose, D-fructofuranonse, D-xylopyranose, D-galactopyranose and D-ribose), ethyl glucoside and 3,4-(methylenedioxy) mandelic acid could dock directly into the active site of COX1. ConclusionAlditols and monosaccharides from sorghum vinegar inhibit multiple steps in the platelet aggregation pathway, and may be beneficial for the treatment of cardiovascular diseases.

Mo1783 Class III PI3K (HVPS34) Is Involved in AMPK-Regulated Neurotensin Secretion Independent of mTORC1 Signaling

Class III PI3K, also known as hVps34, along with its associated regulatory subunit, hVps15 kinase, are critical components of nutrient sensing, and act as upstream factors required for activation of S6K1, a downstream effector of mTOR complex 1 (mTORC1). The hVps34/ hVps15 complex regulates a variety of cellular functions such as intracellular vesicle trafficking. In addition, AMPK regulates the hVps34-containing complex in response to changes in energy state. Neurotensin (NT), a gut hormone produced and stored in N cells of the distal small bowel, has multiple physiologic effects, including facilitation of fatty acid translocation from the intestinal lumen, coordination of gut motility and stimulation of pancreaticobiliary secretion. Since we have previously found that AMPK and mTORC1 positively and negatively regulate NT release, respectively, the purpose of the present study was to investigate whether hVps34 and hVps15 are required in AMPK or mTORC1-mediated secretion. METHODS. i) The human endocrine cell line, BON, was used for all experiments. BON cells synthesize and secrete NT peptide and process the NT peptide in a manner analogous to that of N cells in the small bowel, thus serving as a useful model for enteroendocrine cell secretion. AMPK activation was induced by Aicar or 2-DG, and NT secretion measured using an NT EIA kit. ii) To determine the involvement of hVps34 and hVps15 in AMPK-mediated effects on NT secretion, endogenous expression of hVps34 and hVps15 was inhibited by siRNA-mediated knockdown. Stable BON cell lines overexpressing either the control vector or hVps34/hVps15 (hVps34 and hVps15 are expressed in one vector but regulated by individual promoters) were established. iii) The physical association of hVps34 with AMPK was evaluated by co-immunoprecipitation (co-IP). RESULTS. NT secretion (induced by Aicar or 2-DG) was attenuated in BON cells transfected with siRNAs targeting hVps34 and hVps15 compared to non-targeting control (NTC) siRNA. Overexpression of hVps34/hVps15 increased both basal and Aicarand 2-DG-stimulated NT secretion. Moreover, we detected the physical association of hVps34 with AMPK by co-IP. hVps34 was detected in both AMPKα1and α2-precipitated complexes with increased hVps34 bound in the AMPKα2 complex. CONCLUSIONS. These results identify a novel regulatory mechanism for the hVps34/hVps15 complex in NT peptide secretion, providing a direct connection between AMPK and hVps34 which is independent of mTORC1/S6K1 signaling.

Mo1787 Valproic Acid (VPA), a Histone Deacetylase Inhibitor That Reduces Intraabdominal Adhesions Modulates Peritoneal Plasma Extravasation and Genes That Regulate Fibrin Deposition and Stability

Class III PI3K, also known as hVps34, along with its associated regulatory subunit, hVps15 kinase, are critical components of nutrient sensing, and act as upstream factors required for activation of S6K1, a downstream effector of mTOR complex 1 (mTORC1). The hVps34/ hVps15 complex regulates a variety of cellular functions such as intracellular vesicle trafficking. In addition, AMPK regulates the hVps34-containing complex in response to changes in energy state. Neurotensin (NT), a gut hormone produced and stored in N cells of the distal small bowel, has multiple physiologic effects, including facilitation of fatty acid translocation from the intestinal lumen, coordination of gut motility and stimulation of pancreaticobiliary secretion. Since we have previously found that AMPK and mTORC1 positively and negatively regulate NT release, respectively, the purpose of the present study was to investigate whether hVps34 and hVps15 are required in AMPK or mTORC1-mediated secretion. METHODS. i) The human endocrine cell line, BON, was used for all experiments. BON cells synthesize and secrete NT peptide and process the NT peptide in a manner analogous to that of N cells in the small bowel, thus serving as a useful model for enteroendocrine cell secretion. AMPK activation was induced by Aicar or 2-DG, and NT secretion measured using an NT EIA kit. ii) To determine the involvement of hVps34 and hVps15 in AMPK-mediated effects on NT secretion, endogenous expression of hVps34 and hVps15 was inhibited by siRNA-mediated knockdown. Stable BON cell lines overexpressing either the control vector or hVps34/hVps15 (hVps34 and hVps15 are expressed in one vector but regulated by individual promoters) were established. iii) The physical association of hVps34 with AMPK was evaluated by co-immunoprecipitation (co-IP). RESULTS. NT secretion (induced by Aicar or 2-DG) was attenuated in BON cells transfected with siRNAs targeting hVps34 and hVps15 compared to non-targeting control (NTC) siRNA. Overexpression of hVps34/hVps15 increased both basal and Aicarand 2-DG-stimulated NT secretion. Moreover, we detected the physical association of hVps34 with AMPK by co-IP. hVps34 was detected in both AMPKα1and α2-precipitated complexes with increased hVps34 bound in the AMPKα2 complex. CONCLUSIONS. These results identify a novel regulatory mechanism for the hVps34/hVps15 complex in NT peptide secretion, providing a direct connection between AMPK and hVps34 which is independent of mTORC1/S6K1 signaling.

Mo1786 HIF-1α Plays a Gut-Injurious Role in Interferon-γ Induced Intestinal Epithelial Barrier Dysfunction

Class III PI3K, also known as hVps34, along with its associated regulatory subunit, hVps15 kinase, are critical components of nutrient sensing, and act as upstream factors required for activation of S6K1, a downstream effector of mTOR complex 1 (mTORC1). The hVps34/ hVps15 complex regulates a variety of cellular functions such as intracellular vesicle trafficking. In addition, AMPK regulates the hVps34-containing complex in response to changes in energy state. Neurotensin (NT), a gut hormone produced and stored in N cells of the distal small bowel, has multiple physiologic effects, including facilitation of fatty acid translocation from the intestinal lumen, coordination of gut motility and stimulation of pancreaticobiliary secretion. Since we have previously found that AMPK and mTORC1 positively and negatively regulate NT release, respectively, the purpose of the present study was to investigate whether hVps34 and hVps15 are required in AMPK or mTORC1-mediated secretion. METHODS. i) The human endocrine cell line, BON, was used for all experiments. BON cells synthesize and secrete NT peptide and process the NT peptide in a manner analogous to that of N cells in the small bowel, thus serving as a useful model for enteroendocrine cell secretion. AMPK activation was induced by Aicar or 2-DG, and NT secretion measured using an NT EIA kit. ii) To determine the involvement of hVps34 and hVps15 in AMPK-mediated effects on NT secretion, endogenous expression of hVps34 and hVps15 was inhibited by siRNA-mediated knockdown. Stable BON cell lines overexpressing either the control vector or hVps34/hVps15 (hVps34 and hVps15 are expressed in one vector but regulated by individual promoters) were established. iii) The physical association of hVps34 with AMPK was evaluated by co-immunoprecipitation (co-IP). RESULTS. NT secretion (induced by Aicar or 2-DG) was attenuated in BON cells transfected with siRNAs targeting hVps34 and hVps15 compared to non-targeting control (NTC) siRNA. Overexpression of hVps34/hVps15 increased both basal and Aicarand 2-DG-stimulated NT secretion. Moreover, we detected the physical association of hVps34 with AMPK by co-IP. hVps34 was detected in both AMPKα1and α2-precipitated complexes with increased hVps34 bound in the AMPKα2 complex. CONCLUSIONS. These results identify a novel regulatory mechanism for the hVps34/hVps15 complex in NT peptide secretion, providing a direct connection between AMPK and hVps34 which is independent of mTORC1/S6K1 signaling.

Public Health Implications of Cryptococcal Infection among HIV Patients on Antiretroviral Therapy in Hospital in Shika, Nigeria

Cryptococcal meningitis is a leading cause of death in AIDS patients and contributes substantially to the high early mortality in antiretroviral treatment (ART) programs in low-resource settings. A hospital-based seroprevalence of cryptococcal antigen among HIV patients on ART program was determined using the Alpha cryptococccal Antigen EIA KIT. The prevalence was low in the study area probably because the ART monitoring program used helped in reducing the incidence of cryptococccal infection. Routine screening of HIV patients is needed for early detection of the infection before unset of complicating clinical signs.

Expression of SDF-1 and CXCR4 transcript variants and CXCR7 in epithelial ovarian cancer.

Chemokine stromal cell-derived factor-1 (SDF-1) and its receptors, CXCR4 and CXCR7, have been implicated in epithelial ovarian cancer progression and metastasis. However, limited data are available on the expression levels of SDF-1 and CXCR4 variants and CXCR7 in human epithelial ovarian cancer. The present study aimed to characterize the expression pattern and levels of SDF-1, CXCR4 and CXCR7 in normal human ovaries and epithelial ovarian cancer. The expression of SDF-1 and CXCR4 transcript variants and CXCR7 was determined by quantitative polymerase chain reaction (qPCR). Plasma SDF-1α levels were determined by commercially available EIA kits and cancer antigen 125 (CA 125) levels were quantified by automated microparticle enzyme immunosorbent assay. High expression levels of SDF-1 transcript variant 1 were identified in ovarian cancer and control ovaries. By contrast, in both groups the expression levels of SDF-1 transcript variants 3 and 4 were extremely low. Furthermore, SDF-1 variant 1 levels were notably higher in epithelial ovarian cancer than in control ovaries, while data for the remaining transcripts were similar in both groups. CXCR4 transcript variant 2 and CXCR7 expression levels in normal and neoplastic ovaries were similar. In both groups, CXCR4 transcript variant 2 was not detected. Plasma SDF-1α levels were notably higher in females with epithelial ovarian cancer than in the control ovaries. Elevated levels of blood SDF-1α were found prior to surgery, 6 days after surgery and following completion of the first chemotherapy course. These increases were independent of the type of epithelial ovarian cancer. Our results suggest that the expression of SDF-1 and the genes controlling alternative splicing are elevated in epithelial ovarian cancer, leading to an increased formation of SDF-1 variant 1. Elevated plasma SDF-1α levels in epithelial ovarian cancer patients are not associated with the presence of tumors and/or metastases, however reflect a general response to the disease.

Mechanisms of human eosinophil migration induced by the combination of IL-5 and the endocannabinoid 2-arachidonoyl-glycerol

Mechanisms of human eosinophil migration induced by the combination of IL-5 and the endocannabinoid 2-arachidonoyl-glycerol

Serum concentration of bone morphogenetic proteins (BMPs) is not linked to serum anti-mullerian hormone (AMH) level

Serum concentration of anti-mullerian hormone (AMH) is used as a biomarker in practical clinics. As bone morphogenetic protein (BMP) cytokines induce AMH expression in human granulosa cells (GC) in vitro, serum concentrations of BMP cytokines and AMH were evaluated whether there would be a relationship between BMP cytokines and AMH. Serum concentrations of BMP-2, -6 -7, AMH and FSH were measured in 25 infertility patients using EIA or ELISA kits. Among 25 infertile patients, serum BMP-2, -6, and -7 were detected only in 10, 3, 7 patients respectively, while AMH was detected in 24 out of 25 patients. There was no relation between BMP cytokines and AMH concentration in serum. The detection rate of these BMPs in serum was much lower than that of AMH. Serum concentration of BMP-2, -6, -7 could not estimate serum AMH level.

Vitamin D serum levels and vitamin D receptor FokI polymorphism on tuberculosis children in Palembang, Indonesia

Background Deficiency vitamin D and vitamin D receptor (VDR) polymorphism are strongly associated with the susceptibility of tuberculosis in four seasons countries. As a country with sufficient sunlight, the incidence of tuberculosis in Indonesia remains high. Objective To asses the association between vitamin D serum levels and VDR FokI polymorphisms with the incidence of tuberculosis in children at Palembang. Methods A case control study was conducted at Department of Child Health Mohammad Hoesin Hospital Palembang on November 2011 to April 2012. Group samples consisted of children with tuberculosis based on inclusion criteria. Vitamin D (1.25(OH)2D3/calcitriol) serum level was measured by using IDS 1,25-Dihydroxy Vitamin D EIA kit and VDR FokI polymorphism was identified through RFLP analysis. Results Sixty subjects was divided equally to case and control groups. The mean of calcitriol serum level in case group was lower compared to control which are 105.5 (SD 66.9) pmol/L versus 162.9 (SD 52.9) pmol/L (P=0.001). We found nine subjects with calcitriol deficiency, eight out of thirty in children with tuberculosis and one out of thirty in healthy contacts (OR 10.5; 95% CI 1.2 to 90.7) pmol/L. The incidence of VDR FokI polymorphism was 28 out of 30 in case group and 22 out of 30 in control group (OR 5,0; 95% CI 0,9 to 26,4). Conclusion Vitamin D (calcitriol) deficiency and lower serum level are associated with higher risk of tuberculosis in children at Palembang. Polymorphism FokI in VDR gene also contributes to the susceptibility of tuberculosis.

The expression of angiotensin-converting enzyme 2–angiotensin-(1–7)–Mas receptor axis are upregulated after acute cerebral ischemic stroke in rats

There is now unequivocal evidence that the angiotensin-converting enzyme 2(ACE2)–Ang-(1–7)–Mas axis is a key component of the renin–angiotensin system (RAS) cascade, which is closely correlated with ischemic insult occurrence. Our previous studies demonstrated that the Ang-(1–7), was an active member of the brain RAS. However, the ACE2–Ang-(1–7)–Mas axis expression after cerebral ischemic injury are currently unclear. In the present study, we investigated the time course of ACE2–Ang-(1–7) and Mas receptor expression in the acute stage of cerebral ischemic stroke. The content of Ang-(1–7) in ischemic tissues and blood serum was measured by specific EIA kits. Real-time PCR and western blot were used to determine messenger RNA (mRNA) and protein levels of the ACE2 and Mas. The cerebral ischemic lesion resulted in a significant increase of regional cerebral and circulating Ang-(1–7) at 6–48h compared with sham operation group following focal ischemic stroke (12h: 7.276±0.320ng/ml vs. 2.466±0.410ng/ml, serum; 1.024±0.056ng/mg vs. 0.499±0.032, brain) (P<0.05). Both ACE2 and Mas expression were markedly enhanced compared to the control in the ischemic tissues (P<0.05). Mas immunopositive neurons were also seen stronger expression in the ischemic cortex (19.167±2.858 vs. 7.833±2.483) (P<0.05). The evidence collected in our present study will indicate that, ACE2–Ang-(1–7)–Mas axis are upregulated after acute ischemic stroke and would play a pivotal role in the regulation of acute neuron injury in ischemic cerebrovascular diseases.

Oxytocin precursor gene expression in bovine skeletal muscle is regulated by 17β-oestradiol and dexamethasone

Growth promoter administration, in livestock, potentially poses a major threat to public health, due to the potential endocrine and carcinogenic activity of residues, accumulating in edible tissues, such as skeletal muscle. Therefore, development of new screening tests and methods for the detection of illicit treatments of food animals would be useful. In this study the serum concentrations of oxytocin peptide were measured in beef cattle receiving 17β oestradiol, dexamethasone or placebo over a period of 40days. Changes in gene expression of oxytocin precursor in skeletal muscle were also examined in these animals. Serum analysis using an oxytocin EIA kit indicated a significant up-regulation of the biosynthesis of this nonapeptide only in cattle after 17β oestradiol, but not after dexamethasone or placebo treatment. Quantitative PCR (qPCR) analysis showed a significant overexpression of the oxytocin precursor gene by 33.5 and 13.3-fold in cattle treated with 17β oestradiol and dexamethasone, respectively, in comparison to placebo treated animals. Regulation of gene expression by some myogenic regulatory factors in skeletal muscle was also evaluated in these animal groups, confirming the activity of both growth promoters on this gene. To investigate the use of the oxytocin precursor gene as biomarker for 17β oestradiol and dexamethasone treatment in beef cattle, an absolute quantification of this gene by qPCR was developed. A standard curve was generated and developed with TaqMan® technology and optimal criterion value, sensitivity and specificity of this screening method were established through ROC analysis. This analysis suggested that the up-regulation of oxytocin precursor gene expression in skeletal muscle tissue is a valid marker for detection of illicit 17β oestradiol and/or dexamethasone use in beef cattle. This method may serve as a novel diagnostic tool in the screening phase, and, if introduced in routine testing, may significantly improve overall efficacy and success of the food screening process ordered by state authorities.

The inhibitory effect of a synthetic compound, (Z)-5-(2,4-dihydroxybenzylidene) thiazolidine-2,4-dione (MHY498), on nitric oxide-induced melanogenesis

Nitric oxide (NO) and the NO/PKG signaling pathway play crucial roles in ultraviolet (UV)-induced melanogenesis, which is known to be related to the induction of tyrosinase. In an attempt to find a novel anti-melanogenic agent, we synthesized (Z)-5-(2,4-dihydroxybenzylidene)thiazolidine-2,4-dione (MHY498). The purpose of this study was to investigate the effect of MHY498 on NO levels and on the NO-mediated signaling pathway using an in vitro model of melanogenesis. MHY498 inhibited 200μM sodium nitroprusside (SNP, a NO donor)-induced NO generation, dose-dependently and suppressed tyrosinase activity and melanin synthesis induced by SNP in B16F10 melanoma cells. To investigate the effect of MHY498 on NO-mediated signaling pathway, guanosine cyclic 3′,5′-monophosphate (cGMP) activities were measured using a cGMP EIA Kit and western blotting was performed to determine the effects of MHY498 on the gene expressions of tyrosinase and microphthalmia-associated transcription factor (MITF). The increased activity of cGMP by SNP was reduced dose-dependently by pretreatment with MHY498. Furthermore, MHY498 suppressed the expressions of tyrosinase and MITF stimulated by SNP. This study shows that enhancement of tyrosinase gene expression via the cGMP pathway is a probable primary mechanism of NO-induced melanogenesis and that the NO-mediated signaling pathway with the expression of MITF enhances melanogenesis. In addition, MHY498 was found to scavenge NO and to suppress the activity of the NO-mediated signaling pathway, and thus, to subsequently down-regulate tyrosinase expression and melanogenesis. This study suggests that MHY498 is a promising anti-melanogenic agent that targets the NO-induced cGMP signaling pathway.

Adipocytokines during pregnancy and postpartum in women with gestational diabetes and healthy controls.

This study aims to explore the changes in maternal serum adipocytokines during pregnancy and post partum in normal and complicated with gestational diabetes (GDM) pregnancies and to investigate the relationship between serum adipocytokines and some of major metabolic parameters. 236 pregnant women (127 with GDM and 109 control group) and 50 postpartum women (30 with GDM during pregnancy and 20 controls). Using ELISA and EIA kits, serum levels of adipocytokines were tested during pregnancy and post partum. Maternal adipocytokines levels were correlated with some metabolic parameters. Women with GDM had lower values of adiponectin and higher values of leptin during pregnancy (p<0.001; 0.0001) and post partum (p<0.002; 0.0001). Serum apelin was significantly lower in GDM group (p<0.009). However, we did not find significance for resistin (p<0.317) and apelin (p<0.218). Positive correlation for leptin and negative for adiponectin was found for pre-pregnancy and pregnancy body mass index, glycated hemoglobin and homeostasis model assessment of insulin resistance index. Using cut point of 8.2 μg/ml for adiponectin and 28.7 ng/ml for leptin could exclude GDM with a sensitivity of 83.6%/81.2% and specificity of 56.6%/64.2% (area under the curve 0.702 and 0.827). There are constant differences in adiponectin and leptin levels between GDM and control group during pregnancy and post partum. Apelin was decreased in our GDM group and no differences were found for resistin and visfatin. Further studies are required to verify the mechanism of this alteration and whether the adipocytokines can be predictors for GDM at an early stage of pregnancy.

Confirmation of Reported Aspirin Use in Community Studies

Aspirin (ASA) is recommended for the prevention of cardiovascular disease; however, the compliance is low. Reported use may not reflect actual use. Serum thromboxane B2 (STxB2) measurement was evaluated to validate reported ASA use. Males aged 45 to 79 years and females aged 55 to 79 years completed a survey and STxB2 measurement (Thromboxane B2 EIA Kit; Cayman Chemical, Ann Arbor, Michigan). The 107 patients were grouped by use of ASA (56 ASA+ and 51 ASA-) and possible interfering medications (INT) such as nonsteroidal anti-inflammatory drugs. The STxB2 levels (ng/mL) were significantly lower in ASA users: ASA+ INT- 3.0 (0.7, 8.4), ASA+ INT+ 2.0 (0.8, 4.9), ASA- INT+ 176 (75, 390), and ASA- INT- 271 (199, 366). The INT use did not cause a significant difference in STxB2 levels. A STxB2 cut point of 25 ng/mL had high sensitivity (94.1%) and specificity (91.1%) for ASA use. The STxB2 was a reliable marker of ASA use and could be used to confirm ASA exposure in population-based health studies.

Genetic analysis and epidemiology of Crimean Congo hemorrhagic fever viruses in Baluchistan province of Pakistan

BackgroundPakistan is considered as an endemic country for Crimean-Congo Hemorrhagic fever with numerous outbreaks and sporadic cases reported during the past two decades. Majority of cases are reported from Baluchistan province with subsequent transmissions to non-endemic regions mainly through infected animals directly or via infested ticks. We hereby describe the molecular investigations of CCHF cases reported during 2008 in Quetta city of Baluchistan province.MethodsSerum Samples from 44 patients, with clinical signs of hemorrhagic fever attending a tertiary care hospital in Quetta city, were collected and tested for CCHF virus antigen and genomic RNA, using capture IgM EIA kit and standard RT-PCR assay, respectively. The partial S-gene fragments were directly sequenced to get information related to the prevailing CCHFV genotypes and their molecular epidemiology in Pakistan.ResultsOut of the total forty four, sixteen (36%) samples were found positive for CCHF IgM. Similarly, viral RNA was detected in six (16%) samples. Phylogenetic analysis revealed that all study viruses belong to genotype Asia-1 with closest similarity (99-100%) to the previously reported strains from Pakistan, Afghanistan and Iran.ConclusionWe conclude that CCHF virus remains endemic within Baluchistan and its neighboring regions of Afghanistan warranting a need of incessant surveillance activities.

Ethanol Extract of Dioscorea batatas Stimulates Procollagen Production and Reduces UVB-induced MMPs Activity in Skin

Ultraviolet (UV) B irradiation induces the production of matrix metalloproteinases (MMPs), which are responsible for the degradation or synthesis inhibition of collagenous extracellular matrix in connective tissues, causing skin photoaging. In this study, we examined the inhibitory effect of MMP-1 expression of yam extract in tumor necrosis factor-alpha (TNF-α)-stimulated human dermal fibroblast neonatal (HDFn) cell and preventive effect of UVB-induced damage in hairless mice skin. The synthesis of procollagen and the release of MMP-1 in HDFn cells were measured by EIA kit and MMP-1 assay kit, respectively. UVB radiation was applied to the backs of the mice three times a week for 8 weeks. Mice were randomly divided into three groups, and were topical application with the Dioscorea batatas (DB, 6%) or vehicle. Reduction of TNF-α-induced procollagen synthesis was increased by DB (50 ug/ml), which was higher than positive control group (TGF-β). Also, pre-treatment of HDFn cells with DB inhibited TNF-α?induced release of MMP-1. In vivo study, we found that preventive effect of DB against UV-induced epidermal thickness. DB suppressed the expression of MMP-3 and MMP-13 induced by UVB irradiation. Our results show that DB have preventive effect of UV-induced skin damage in hairless mice.