Mitochondrial dysfunction reactivates α-fetoprotein expression that drives copper-dependent immunosuppression in mitochondrial disease models.

Signaling circuits crucial to systemic physiology are widespread, yet uncovering their molecular underpinnings remains a barrier to understanding the etiology of many metabolic disorders. Here, we identified a copper-linked signaling circuit activated by disruption of mitochondrial function in the murine liver or heart that resulted in atrophy of the spleen and thymus and caused a peripheral white blood cell deficiency. We demonstrated that the leukopenia was caused by α-fetoprotein, which required copper and the cell surface receptor CCR5 to promote white blood cell death. We further showed that α-fetoprotein expression was upregulated in several cell types upon inhibition of oxidative phosphorylation. Collectively, our data argue that α-fetoprotein may be secreted by bioenergetically stressed tissue to suppress the immune system, an effect that may explain the recurrent or chronic infections that are observed in a subset of mitochondrial diseases or in other disorders with secondary mitochondrial dysfunction.

Distal hypospadias repair: Comparative study between snodgrass and transverse preputial onlay flap.

INTRODUCTION: Distal hypospadias repair has several operative strategies such as tubularized incised plate (TIP); Transverse preputial onlay flap (TPOF) and Modified Mathieu Procedure. The aim of our work is to compare between TIP and TPOF techniques in the outcome. MATERIAL AND METHOD: We conducted this comparative study on 66 patients with different types of distal hypospadias divided in two groups A and B for TIP and TPOF respectively. We reported outcome and complications for both TIP and TPOF repair. Our results revealed mean operative time was significantly higher in group B (123.1 ± 6.8 min) than in group A (93.73 ± 3.9 min, P value < 0.001). Postoperative urinary fistula was reported in group A (15.2%) while no fistula in group B (p value = 0.05). The vertical slit appearance of meatus was better in group A (54.5%) than in group B (24.2%, P value 0.023). As regard to hypospadias objective scoring evaluation (HOSE) (Summary Figure) the meatal shape score was higher in group A (1.55 ± 0.51) than in group B (1.24 ± 0.44, p value = 0.0 l), the urinary stream score was significantly higher in group A (1.82 ± 0.39) than in group B (1.97 ± 0.17, p value = 0.04) and the urinary fistula score was significantly higher in group A (3.79 ± 0.60) than in group B (4 ± 0, p value = 0.04). DISCUSSION: Complications are more common after TIP than in TPOF. As regard the meatus shape, the naturally looking vertical slit like meatus was achieved more in the TIP than in TPOF. We advocate further studies with a large number of patients with a long follow up period. CONCLUSION: TPOF is a great option to consider in cases of distal hypospadias because it is an effective technique for primary distal hypospadias repair with a good cosmetic outcome and a low associated complication.

Death of a Protein: The Role of E3 Ubiquitin Ligases in Circadian Rhythms of Mice and Flies.

Circadian clocks evolved to enable organisms to anticipate and prepare for periodic environmental changes driven by the day-night cycle. This internal timekeeping mechanism is built on autoregulatory transcription-translation feedback loops that control the rhythmic expression of core clock genes and their protein products. The levels of clock proteins rise and ebb throughout a 24-h period through their rhythmic synthesis and destruction. In the ubiquitin-proteasome system, the process of polyubiquitination, or the covalent attachment of a ubiquitin chain, marks a protein for degradation by the 26S proteasome. The process is regulated by E3 ubiquitin ligases, which recognize specific substrates for ubiquitination. In this review, we summarize the roles that known E3 ubiquitin ligases play in the circadian clocks of two popular model organisms: mice and fruit flies. We also discuss emerging evidence that implicates the N-degron pathway, an alternative proteolytic system, in the regulation of circadian rhythms. We conclude the review with our perspectives on the potential for the proteolytic and non-proteolytic functions of E3 ubiquitin ligases within the circadian clock system.

Vitamin D3 enhances the effects of omega-3 oils against metabolic dysfunction-associated fatty liver disease in rat.

This study investigated the effects of omega-3 oils (OM) and/or vitamin D3 (VD) against metabolic dysfunction-associated fatty liver disease (MAFLD). Forty rats were divided into negative (NC) and positive (PC) controls, OM, VD, and OM + VD groups, and MAFLD was induced by high-fat/high-fructose diet (12 weeks). Oral OM (415 mg/kg/day) and/or intramuscular VD (290 IU/kg/day) were given for 4 weeks (5 times/week). The PC animals were markedly obese and had hyperglycemia, insulin resistance, dyslipidemia, elevated liver enzymes, abnormal hepatic histology, and increased caspase-3 with apoptosis than the NC group. The expression of hepatic peroxisome proliferator-activated receptor-α (PPAR-α; 5.3-fold), insulin induced gene-1 (INSIG1; 7.8-fold), adiponectin receptor-1 (AdipoR1; 4.4-fold), and leptin receptor (LEPR; 6-fold) declined, while PPAR-γ (3.7-fold) and sterol regulatory element-binding protein-1 (SREBP1; 2.4-fold) increased, in the PC than the NC group. Leptin (2.2-fold), malondialdehyde (2.1-fold), protein carbonyl groups (17.3-fold), IL-1ß (4.4-fold), IL-6 (2.1-fold), TNF-α (1.8-fold) also increased, whereas adiponectin (2.8-fold) glutathione (2.1-fold), glutathione peroxidase-1 (2.4-fold), glutathione reductase (2.2-fold), catalase (1.4-fold), and IL-10 (2.8-fold) decreased, in the PC livers. Both monotherapies attenuated obesity, metabolic profiles, and PPAR-γ/SREBP1/leptin/Caspase-3/apoptosis, while induced PPAR-α/adiponectin/AdipoR1/LEPR/INSIG1. The monotherapies also reduced the oxidative stress and pro-inflammatory markers and increased the antioxidant and anti-inflammatory molecules. However, the OM effects were better than VD monotherapy. Alternatively, the co-therapy group showed the greatest ameliorations in liver functions, lipid-regulatory molecules, oxidative stress, inflammation, and apoptosis. In conclusion, while OM monotherapy was superior to VD, the co-therapy protocol displayed the best alleviations against MAFLD, possibly by enhanced modulation of metabolic, antioxidant, and anti-inflammatory pathways.

Genetic Variability and Evidence of a New Subgroup in Watermelon Mosaic Virus Isolates.

Watermelon mosaic virus (WMV) is one of the important Potyviruses that infect cucurbits worldwide. To better understand the population structure of WMV in the United States (U.S.), 57 isolates were collected from cucurbit fields located in nine southern states. The complete coat protein gene of all WMV isolates was cloned, sequenced and compared with 89 reported WMV isolates. The nucleotide and amino acid sequence identities among the U.S. WMV isolates ranged from 88.9 to 99.7% and from 91.5 to 100%, respectively. Phylogenetic analysis revealed that all the U.S. WMV isolates irrespective of their geographic origin or hosts belonged to Group 3. However, the fifty-seven isolates made three clusters in G3, where two clusters were similar to previously reported subgroups EM1 and EM2, and the third cluster, containing nine WMV isolates, formed a distinct subgroup named EM5 in this study. The ratio of non-synonymous to synonymous nucleotide substitution was low indicating the occurrence of negative purifying selection in the CP gene of WMV. Phylogenetic analysis of selected 37 complete genome sequences of WMV isolates also supported the above major grouping. Recombination analysis in the CP genes confirmed various recombinant events, indicating that purifying selection and recombination are the two dominant forces for the evolution of WMV isolates in the U.S.

Effects of dapagliflozin in combination with insulin on cytochrome P450 activities in a diabetes type 1 rat model.

Previous studies reported that diabetes alters the activities of hepatic cytochrome P450 (CYP) enzymes, which, in turn, affects the disposition of some drugs. We herein examined and compared the effects of the combination of dapagliflozin with a low insulin dose, a full dose of insulin alone, and dapagliflozin alone for 3 and 8 weeks on CYP activities in a diabetes type 1 rat model. We induced type 1 diabetes in rats using a single intraperitoneal injection of 60 mg/kg streptozotocin (STZ). Daily treatment with the full dose of insulin alone, dapagliflozin alone, or dapagliflozin in combination with a low dose of insulin was then initiated. STZ-induced rats developed marked hyperglycemia and altered CYP2E activities. Dapagliflozin in combination with a low dose of insulin stabilized hyperglycemia and CYP1A, 2D, 2E and 3A activities. However, dapagliflozin alone did not improve blood glucose levels or CYP activities. These results suggest that the effects of dapagliflozin in combination with a low dose of insulin are similar to those of a full dose of insulin, and stabilize CYP activities in type 1 diabetes.

Failure Mechanism of Hybrid Steel Beams with Trapezoidal Corrugated-Web Non-Welded Inclined Folds.

Literature of Steel Beams with a thin-walled trapezoidal Corrugated Web (SBCWs) shows that the capacity of SBCWs is affected by both the fatigue cracks initiated along the inclined folds (IFs) and the maximal additional stress located in the middle of the IFs. An experimental investigation on the behaviour of hybrid SBCWs under flexure is presented in this paper. This study focuses on the effect of the welding IF between the web and flanges (IFs welded or non-welded), the horizontal-fold length (200, 260, and 350 mm), and transversal flange stiffeners on the failure mechanism of the SBCW under three line load. Accordingly, six hybrid specimens were fabricated, instrumented, and tested (five SBCW specimens and one specimen with a flat web). The test setup was designed to generate shear and a moment in the testing zone via three-point bending. The results indicated that non-welded IFs specimens with or without flange stiffeners failed owing to web tearing after web and flange local buckling. The failure mode of the specimen with continuous welding between the web and flanges was local flange buckling. Finally, the paper presents a comparison between the experimental results and the European Code to predict the capacity of the flange towards local buckling. It was concluded that the non-welding the IFs affected the inelastic behaviour and the capacity of the SBCWs. In addition, the bending resistance equations presented by EN 1993-1-5 can safely predict the test results of the non-welded inclined fold and yield a high safe variation.

SOX2 Regulates Neuronal Differentiation of the Suprachiasmatic Nucleus.

In mammals, the hypothalamic suprachiasmatic nucleus (SCN) functions as the central circadian pacemaker, orchestrating behavioral and physiological rhythms in alignment to the environmental light/dark cycle. The neurons that comprise the SCN are anatomically and functionally heterogeneous, but despite their physiological importance, little is known about the pathways that guide their specification and differentiation. Here, we report that the stem/progenitor cell transcription factor, Sex determining region Y-box 2 (Sox2), is required in the embryonic SCN to control the expression of SCN-enriched neuropeptides and transcription factors. Ablation of Sox2 in the developing SCN leads to downregulation of circadian neuropeptides as early as embryonic day (E) 15.5, followed by a decrease in the expression of two transcription factors involved in SCN development, Lhx1 and Six6, in neonates. Thymidine analog-retention assays revealed that Sox2 deficiency contributed to reduced survival of SCN neurons during the postnatal period of cell clearance, but did not affect progenitor cell proliferation or SCN specification. Our results identify SOX2 as an essential transcription factor for the proper differentiation and survival of neurons within the developing SCN.

Effect of dapagliflozin alone and in combination with insulin in a rat model of type 1 diabetes.

Dapagliflozin is a selective sodium-glucose cotransporter 2 (SGLT2) inhibitor; it reduces glucose reabsorption via the kidney and increases the glucose excretion in urine. This inhibitor functions through a unique insulin-independent mechanism, and is therefore a potential new approach for the treatment of hyperglycemia in patients with diabetes. In this study, we evaluated the effectiveness of the SGLT2 inhibitor, dapagliflozin, by using a rat model of type 1 diabetes. Type 1 diabetes was induced by a single intraperitoneal injection of 60 mg/kg streptozotocin (STZ). The STZ-induced rats showed marked hyperglycemia and other metabolic abnormalities. We clarified the hypoglycemic effect of the combination treatment of dapagliflozin with a low dose of insulin compared with dapagliflozin alone and insulin alone in 3-week and 8-week studies. Our results showed that dapagliflozin in combination with a low dose of insulin significantly lowered hyperglycemia, hypercholesterolemia, and hypertriglyceridemia. Furthermore, the antioxidant status and body weight were improved. In contrast, treatment with dapagliflozin alone did not improve the blood glucose levels, lipid profile, antioxidant status, or body weight. These findings suggested that in type 1 diabetes, dapagliflozin was effective in combination with a low dose of insulin; however, the administration of dapagliflozin alone did not achieve a significant effect.

Predictors of In-Hospital Mortality in Isolated Total Anomalous Pulmonary Venous Connection.

BACKGROUND: We assessed risk factors for early mortality in isolated total anomalous pulmonary venous connection over a modern era excluding emergent cases to eliminate the influence of associated factors on surgical outcome.  Methods: 70 patients with isolated total anomalous pulmonary venous connection who were repaired electively between January 2013 and February 2018 were included. RESULTS: In-hospital mortality was encountered in 4 patients (5.7%). Upon univariate analysis, low age (P = .003) and weight (P = .001) at surgery, preoperative pulmonary venous obstruction (P = .010), preoperative low oxygen saturation (P = .031), long cardiopulmonary bypass (P = .001) and aortic cross clamp (P = .003) times, long duration of mechanical ventilation (P = .001), chest infection (P = 0.041), postoperative low CO syndrome (P < .001) and long postoperative inotropic support (P = .015) were significant predictors of in-hospital mortality. In multivariate analysis postoperative low cardiac output syndrome (OR: 1.060; 95% CI: 1.008-1.116) and prolonged postoperative mechanical ventilation (OR: 1.772; 95% CI: 1.141-2.751) were independent factors of in-hospital mortality. CONCLUSION: Surgical repair of TAPVC is now performed with acceptable results. According to our study, postoperative low cardiac output syndrome and prolonged postoperative mechanical ventilation were the most significant predictors for early mortality.

Protective effects of garlic extract against hematological alterations, immunosuppression, hepatic oxidative stress, and renal damage induced by cyclophosphamide in rats.

Cyclophosphamide is an alkylating agent widely used as anticancer drug, reported to exert cytotoxic effects attributed to oxidative stress. Therefore, this study aimed to explore the protective effect of ethanolic extract of garlic (EEG) against cyclophosphamide (Cyp)-induced hematological disturbance and immunosuppressive and hepatotoxic effects. Forty male Wistar albino rats were randomized into four equal groups: the normal control one, the Cyp-treated group (50 mg/kg BW/IM, once weekly), the EEG-treated group (300 mg/kg BW, orally, daily), and the Cyp & EEG group. All rats received their relevant treatments for four consecutive weeks. This study revealed that Cyp significantly decreased erythrocyte count, hemoglobin (Hb), packed cell volume (PCV), and total leukocyte and lymphocyte counts. However, the counts of neutrophils, eosinophils, and toxic neutrophils were elevated. Additionally, hepatic malondialdehyde (MDA) and levels of liver and renal biomarkers were significantly elevated in the Cyp-treated group. Otherwise, hepatic catalase (CAT), reduced glutathione (GSH), superoxide dismutase (SOD), and serum total antioxidant capacity (TAC) were significantly lower than the control rats. Furthermore, Cyp significantly reduced whole blood respiratory burst activity (NBT), serum lysozyme and bactericidal activities, interlukin-12 (IL-12), and interferon-γ. In contrast, the levels of nitric oxide (NO), tumor necrosis factor-α (TNF-α), and interlukin-1ß (IL-1ß) were elevated. Additionally, Cyp induced hepatic and renal histopathological alterations. Data in the present study demonstrated that EEG has immunomodulatory and antioxidant effects and has the ability to diminish the alterations induced by Cyp.

Enhancement of immune response and resistance against Vibrio parahaemolyticus in kuruma shrimp (Marsupenaeus japonicus) by dietary supplementation of ß-1,4-mannobiose.

Supplementation of prebiotic carbohydrates can act as a potent immunomodulator and have the efficacy to induce immune-related genes which are involved in host defense. Pure ß-1,4-mannobiose (MNB) showed activation of prophenoloxidase system of shrimp hemocytes in vitro. The resistance of kuruma shrimp Marsupenaeus japonicus against Vibrio parahaemolyticus was examined after the shrimp were fed with 0 (control), 0.02, 0.2, and 2% MNB supplemented diets. The results showed significantly higher survival rates in MNB supplemented shrimp than those of the control one from 2 to 12 days post challenge. In another experiment, the hemocyte count, ROS production, phagocytic, phenoloxidase and bactericidal activities, and expression of immune-related genes were investigated in the control and MNB supplemented groups at day 1, 4, 6, 8 and 11 of the feeding. These immune parameters were significantly enhanced in MNB supplemented groups. Furthermore, the gene expression analysis showed that transcripts of lysozyme, crustin, penaeidin and TNF were significantly up-regulated in hemolymph, lymphoid organs and intestines of MNB treated shrimp. Overall, the results provided evidence that MNB supplementation could improve the immune response and increase shrimp resistance against V. parahaemolyticus infection.

Molecular characterization reveals that squash chlorosis mottling virus and zucchini tigré mosaic virus are the same newly emerging potyvirus.

In 2013, we published the first report of a novel potyvirus isolate, which was tentatively named squash chlorosis mottling virus (SqCMV), from an infected squash plant (Cucurbita pepo) collected in the Homestead area of Florida. The purpose of the current work was to further characterize the virus isolate based on molecular properties, and to determine its relationship to other potyviruses. The complete genome sequence of the virus was 10,292 nucleotides (nt), consisting of a 5'-UTR (182 nt), a complete ORF (10,098 nt) encoding a polyprotein of 3,365 amino acids, a motif for the PIPO protein and a 3'-UTR (112 nt), but excluding the poly-(A) tail. Phylogenetic analysis based on the complete genome and amino acid sequences revealed that the virus from Florida clustered with the type isolate of the newly described potyvirus zucchini tigré mosaic virus (ZTMV) sharing 82-90% nucleotide and 83-86% amino acid identities, respectively. Recombination analysis confirmed one major recombination event in the putative P1 coding region of the virus with the putative parental sequences predicted to resemble to ZTMV-Venezuela and France isolates. In addition, genetic diversity analysis indicated that the CP gene was under the highest selection pressure compared to other genes. Together, these results suggest that SqCMV and ZTMV are both representative isolates of the same species, Zucchini tiger mosaic virus. We present the first complete genome sequence of the ZTMV-HFL isolate from the USA.

Prevalence of schistosomiasis among patients attending Qena General Hospital over the years 2011 and 2012.

Schistosomiasis is a public health problem in Egypt. To detect the prevalence of schistosomiasis in Qena Governorate, 1601 urine and stool samples of patients attending Qena General Hospital were examined of whom 1601 patients had urinary symptoms and 893 patients had dysentery and bloody stool. Sheets were filled out on each patient. Also, a total of 7590 Bulinus truncatus and Biomphalaria alexandrina snails were collected from water bodies in Qena Governorate. The overall prevalence of S. haematobium was 13.9%; with maximum among 6-18 year-old age group and higher in males than in females. Risk factors for S. haematobium infection were this age group; particularly males. However, males swim and play in water bodies and women wash cloths and utensils there, and children swimming or playing in canals. The S. haematobium peak of infection was in winter correlated with the highest prevalence in Dandara City, Qena City, Awlad-Amr and El Hijarat. S. mansoni were negative in the examined individuals. The infective rate among snails was 1.82% in B. truncatus and 0.0% among B. alexandrina and M. tuberculata.

Effect of Withania somnifera Extracts on Some Selective Biochemical, Hematological, and Immunological Parameters in Guinea Pigs Experimental Infected with E. coli.

Fifty 1-2-month-old Guinea pigs were divided into 5 equal groups, 10 each. Control (Gp1) did receive neither viable bacteria nor treatment. Each animal from the other groups (Gp2-5) was challenged with (1-2 × 10(8)) viable E. coli in 200 µ L normal saline (0.9%) through IP route. GP2 infected group was treated with 200 µ L saline IP and kept as positive control group. Gp3-4 are infected and treated with Withania somnifera (ethanol root extract) with doses 50 and 100 mg/kg. BW, respectively. Gp5 infected treated group was treated with cefoperazone antibiotic at dose 35 mg/Kg BW. The treatment by drug or the extracted medicinal plant was started 72 h after infection for 7 successive days. Serum and whole blood sample were collected from all groups 14 days after treatment to evaluate some hematological and biochemical changes as well as immunomodulatory cytokine tumor necrosis factor-alpha (TNF- α ). Oral treatment of the plant extract caused significant benefit results in infected Guinea pig appeared in the correction of some hematological and biochemical parameters also try to suppressed inflammatory cytokine response represent in TNF- α . It could be concluded that W. somnifera extract has potent antibacterial activity, and this appears in the correction with hematological, biochemical, and immunological results.

Genetic diversity in the 3'-terminal region of papaya ringspot virus (PRSV-W) isolates from watermelon in Oklahoma.

The 3'-terminal region (1191 nt) containing part of the NIb gene, complete coat protein (CP) and poly-A tail of 64 papaya ringspot virus (PRSV-W) isolates collected during 2008-2009 from watermelon in commercial fields of four different counties of Oklahoma were cloned and sequenced. Nucleotide and amino acid sequence identities ranged from 95.2-100% and 97.1-100%, respectively, among the Oklahoman PRSV-W isolates. Phylogenetic analysis showed that PRSW-W isolates clustered according to the locations where they were collected within Oklahoma, and each cluster contained two subgroups. All subgroups of Oklahoman PRSV-W isolates were on separate branches when compared to 35 known isolates originating from other parts of the world, including the one reported previously from the USA. This study helps in our understanding about the genetic diversity of PRSV-W isolates infecting cucurbits in Oklahoma.