Dose-dependent Manner Research Articles

Doxycycline-Mediated Inhibition of Snake Venom Phospholipase and Metalloproteinase.

Warfighters are exposed to life-threatening injuries daily and according to the Joint Trauma System Military Clinical Practice Guideline-Global Snake Envenomation Management snakebites are a concerning threat in all theaters of operation. Snake venom is a complex mixture of toxins including phospholipases A2 (PLA2) and snake venom metalloproteinases (SVMP) that produce myotoxic, hemotoxic, and cytotoxic injuries. Antibody-based antivenom is the standard of care but new approaches including small-molecule inhibitors have gained attention in recent years. Doxycycline is an effective inhibitor of human metalloproteinases and PLA2. The enzymatic activities of 3 phylogenetically distinct snakes: Agkistrodon piscivorus, Naja kaouthia, and Daboia russelii were tested under inhibitory conditions using doxycycline. Enzymatic activity of PLA2 and SVMP was measured in N. kaouthia, D. russelii, and A. piscivorus venom alone and with doxycycline using EnzChek Phospholipase A2 and Gelatinase Assay Kits. A 1-way ANOVA with Tukey's post-hoc test was used to conduct comparative analysis. The median lethal dose of the venoms, the effective dose of doxycycline, and creatine kinase (CK) inhibition levels were measured in a murine model with adult Bagg Albino (BALB/c) mice using intramuscular injections. Median lethal and effective doses were determined using Spearman-Karber's method and a 1-way ANOVA with Tukey's post-hoc test was used to compare CK inhibition levels. Phospholipases A2 activity was reduced to 1.5% to 44.0% in all 3 venoms in a dose-dependent manner using 0.32, 0.16, and 0.08 mg/mL doxycycline when compared to venom-only controls (P < .0001) (Fig.1A). Snake venom metalloproteinases activity was reduced to 4% to 62% in all 3 venoms in a dose-dependent manner using 0.32, 0.16, and 0.08 mg/mL doxycycline (P < .0001) (Fig.1B). The lethal dose (LD50) values of the venoms in the murine model were calculated as follows: A. piscivorus = 20.29 mg/kg (Fig.2A), N. kaouthia = 0.38 mg/kg (Fig.2B), and D. russelii = 7.92 mg/kg (Fig.2C). The effective dose (ED50) of doxycycline in A. piscivorus was calculated to be 20.82 mg/kg and 72.07 mg/kg when treating D. russelii venom. No ED50 could be calculated when treating N. kaouthia venom (Fig.3). Creatine kinase activity was significantly decreased in all 3 venoms treated with doxycycline (P < .0001) (Fig.4). Doxycycline reduced PLA2- and SVMP-related lethality, particularly in A. piscivorus envenomings and in a limited capacity with D. russelii revealing its promise as a treatment for snakebites. In addition, CK activity, a common indicator of muscle damage was inhibited in mice that received doxycycline-treated venom. The doxycycline concentrations identified in the ED50 studies correspond to 1,456 to 5,061 mg dosages for a 70 kg human. Factors including venom yield and snake species would affect the actual dosage needed. Studies into high-dose doxycycline safety and its effectiveness against several snake species is needed to fully translate its use into humans. Based on this work, doxycycline could be used as a treatment en route to higher echelons of care, providing protection from muscle damage and reducing lethality in different snake species.

Fillable Magnetic Microrobots for Drug Delivery to Cardiac Tissues in Vitro.

Many cardiac diseases, such as arrhythmia or cardiogenic shock, cause irregular beating patterns that must be regulated to prevent disease progression towards heart failure. Treatments can include invasive surgery or high systemic drug dosages, which lack precision, localization, and control. Drug delivery systems (DDSs) that can deliver cargo to the cardiac injury site could address these unmet clinical challenges. Here, we present a microrobotic DDS that can be mobilized to specific sites via magnetic control. Our DDS incorporates an internal chamber that can protect drug cargo. Furthermore, the DDS contains a tunable thermosensitive sealing layer that gradually degrades upon exposure to body temperature, enabling prolonged drug release. Once loaded with the small molecule drug norepinephrine, our microrobotic DDS modulated beating frequency in induced pluripotent stem-cell derived cardiomyocytes (iPSC-CMs) in a dose-dependent manner, thus simulating drug delivery to cardiac cells in vitro. The DDS also navigated several maze-like structures seeded with cardiomyocytes to demonstrate precise locomotion under a rotating low-intensity magnetic field and on-site drug delivery. This work demonstrates the utility of a magnetically actuating DDS for precise, localized, and controlled drug delivery which is of interest for a myriad of future opportunities such as in treating cardiac diseases. This article is protected by copyright. All rights reserved.

NF-κB factors cooperate with Su(Hw)/E4F1 to balance Drosophila/human immune responses via modulating dynamic expression of miR-210.

MicroRNAs (miRNAs) play crucial regulatory roles in controlling immune responses, but their dynamic expression mechanisms are poorly understood. Here, we firstly confirm that the conserved miRNA miR-210 negatively regulates innate immune responses of Drosophila and human via targeting Toll and TLR6, respectively. Secondly, our findings demonstrate that the expression of miR-210 is dynamically regulated by NF-κB factor Dorsal in immune response of Drosophila Toll pathway. Thirdly, we find that Dorsal-mediated transcriptional inhibition of miR-210 is dependent on the transcriptional repressor Su(Hw). Mechanistically, Dorsal interacts with Su(Hw) to modulate cooperatively the dynamic expression of miR-210 in a time- and dose-dependent manner, thereby controlling the strength of Drosophila Toll immune response and maintaining immune homeostasis. Fourthly, we reveal a similar mechanism in human cells, where NF-κB/RelA cooperates with E4F1 to regulate the dynamic expression of hsa-miR-210 in the TLR immune response. Overall, our study reveals a conservative regulatory mechanism that maintains animal innate immune homeostasis and provides new insights into the dynamic regulation of miRNA expression in immune response.

Chemical Derivatization Leads to the Discovery Of Novel Analogs of Azotochelin, a Natural Siderophore, as Promising Anticancer Agents.

Siderophores are structurally unique medicinal natural products and exhibit considerable therapeutic potential. Herein, we report the design and synthesis of azotochelin, a natural siderophore, and an extensive library of azotochelin analogs and their anticancer properties. We modified the carboxylic acid and the aromatic ring of azotochelin using various chemical motifs. We evaluated the cytotoxicity of the compounds against six different cancer cell lines (KB-3-1, SNB-19, MCF-7, K-562, SW-620, and NCI-H460) and a non-cancerous cell line (HEK-293). Among the twenty compounds tested, the IC50 values of nine compounds (14, 32, 35-40, and 54) were between 0.7 and 2.0 μM against a lung cancer cell line (NCI-H460). Moreover, several compounds showed good cytotoxicity profile (IC50 <10 μM) against the tested cancer cell lines. The flow cytometry analysis showed that compounds 36 and 38 induced apoptosis in NCI-H460 in a dose-dependent manner. The cell cycle analysis indicated that compounds 36 and 38 significantly arrested the cell cycle at the S phase to block cancer cell proliferation in the NCI-H460 cell line. The study has produced various novel azotochelin analogs that are potentially effective anticancer agents and lead compounds for further synthetic and medicinal chemistry exploration.

A mutation in the PRKAR1B gene drives pathological mechanisms of neurodegeneration across species.

Protein Kinase A (PKA) neuronal function is controlled by the interaction of a regulatory (R) subunit dimer to two catalytic (C) subunits. Recently, the L50R variant in the gene encoding the RIβ subunit was identified in individuals with a novel neurodegenerative disease. However, the mechanisms driving the disease phenotype remained unknown. In this study, we generated a mouse model carrying the RIβ-L50R mutation to replicate the human disease phenotype and study its progression with age. We examined postmortem brains of affected individuals as well as live cell cultures. Employing biochemical assays, immunohistochemistry, and behavioral assessments, we investigated the impact of the mutation on PKA complex assembly, protein aggregation and neuronal degeneration. We reveal that RIβ is an aggregation-prone protein that progressively accumulates in wildtype and Alzheimer's mouse models with age, while aggregation is accelerated in the RIβ-L50R mouse model. We define RIβ-L50R as a causal mutation driving an age-dependent behavioral and disease phenotype in human and mouse models. Mechanistically, this mutation disrupts RIβ dimerization, leading to aggregation of its monomers. Intriguingly, interaction with the C-subunit protects the RIβ-L50R from self-aggregating, in a dose-dependent manner. Furthermore, cAMP signaling induces RIβ-L50R aggregation. The pathophysiological mechanism elucidated here for a newly recognized neurodegenerative disease, in which protein aggregation is the result of disrupted homodimerization, sheds light on a remarkably under-appreciated but potentially common mechanism across several neurodegenerative diseases.

Serotonin Promotes Vesicular Association and Fusion by Modifying Lipid Bilayers.

The primary event in chemical neurotransmission involves the fusion of a membrane-limited vesicle at the plasma membrane and the subsequent release of its chemical neurotransmitter cargo. The cargo itself is not known to have any effect on the fusion event. However, amphiphilic monoamine neurotransmitters (e.g., serotonin and dopamine) are known to strongly interact with lipid bilayers and to affect their mechanical properties, which can in principle impact membrane-mediated processes. Here, we probe whether serotonin can enhance the association and fusion of artificial lipid vesicles in vitro. We employ fluorescence correlation spectroscopy and total internal reflection fluorescence microscopy to measure the attachment and fusion of vesicles whose lipid compositions mimic the major lipid components of synaptic vesicles. We find that the association between vesicles and supported lipid bilayers is strongly enhanced in a serotonin dose-dependent manner, and this drives an increase in the rate of spontaneous fusion. Molecular dynamics simulations and fluorescence spectroscopy data show that serotonin insertion increases the water content of the hydrophobic part of the bilayer. This suggests that the enhanced membrane association is likely driven by an energetically favorable drying transition. Other monoamines, such as dopamine and norepinephrine, but not other related species, such as tryptophan, show similar effects on membrane association. Our results reveal a lipid bilayer-mediated mechanism by which monoamines can themselves modulate vesicle fusion, potentially adding to the control toolbox for the tightly regulated process of neurotransmission in vivo.

The effect of Phyllanthus emblica (Amla) fruit supplementation on the rumen microbiota and its correlation with rumen fermentation in dairy cows

IntroductionMedicinal plants, rich in phytochemicals like phenolic acids, flavonoids, and tannins, offer potential benefits in enhancing productivity, quality, and animal health. Amla fruit (Phyllanthus emblica) is one such plant with promising attributes. This study aimed to investigate the impact of fresh Amla fruit (FAF) supplementation on ruminal microbial composition and its correlation with rumen fermentation in lactating dairy cows.MethodsThe study employed a repeated crossover design involving eight ruminally cannulated mid-lactation Holstein dairy cows. Animals received varying levels of fresh Amla fruit supplementation (0, 200, 400, and 600 g/d).ResultsWhen 400 g/d of FAF was added to the diet, there was a significant increase in the relative abundance of Firmicutes (p = 0.02). However, at 200 g/d, the relative abundance of ruminal Bacteroidota was higher than the 0 and 400 g/d FAF supplementation (p &amp;lt; 0.01). LEfSe analysis identified distinct taxa, such as Clostridia vadinBB60 in the 200 g/d group, Oscillospiraceae in the 400 g/d group, and Elusimicrobium in the 600 g/d group. Notably, the random forest species abundance statistics identified Oscillospiraceae V9D2013 as a biomarker related to milk yield. Oscillospiraceae, Bacilli RF39, norank_f Prevotellaceae, and Bifidobacterium were positively correlated with ruminal total VFA and molar proportion of propionate, while Rikenellaceae RC9 gut group and Clostridia vadinBB60 were negatively correlated.DiscussionFAF supplementation affects the abundance of beneficial microbes in a dose-dependent manner, which can improve milk yield, efficiency, rumen health, desirable fatty acids, and animal health.

Driving Mitochondrial Fission Improves Cognitive, but not Motor Deficits in a Mouse Model of Ataxia of Charlevoix-Saguenay.

Autosomal-recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) is caused by loss-of-function mutation in the SACS gene, which encodes sacsin, a putative HSP70-HSP90 co-chaperone. Previous studies with Sacs knock-out (KO) mice and patient-derived fibroblasts suggested that SACSIN mutations inhibit the function of the mitochondrial fission enzyme dynamin-related protein 1 (Drp1). This in turn resulted in mitochondrial hyperfusion and dysfunction. We experimentally tested this hypothesis by genetically manipulating the mitochondrial fission/fusion equilibrium, creating double KO (DKO) mice that also lack positive (PP2A/Bβ2) and negative (PKA/AKAP1) regulators of Drp1. Neither promoting mitochondrial fusion (Bβ2KO) nor fission (Akap1 KO) influenced progression of motor symptoms in Sacs KO mice. However, our studies identified profound learning and memory deficits in aged Sacs KO mice. Moreover, this cognitive impairment was rescued in a gene dose-dependent manner by deletion of the Drp1 inhibitor PKA/Akap1. Our results are inconsistent with mitochondrial dysfunction as a primary pathogenic mechanism in ARSACS. Instead, they imply that promoting mitochondrial fission may be beneficial at later stages of the disease when pathology extends to brain regions subserving learning and memory.

Aortic Injury Induced by Benzo(a)pyrene and Atherogenic Diet Increased Hepatic FGF21 Expression in C57BL/6J Mice

Background: Benzo(a)pyrene (BaP), an environmental toxicant and endocrine disruptor, has been shown to exacerbate atherosclerosis when combined with a high-fat diet. Fibroblast Growth Factor-21 (FGF21), a novel hormone with anti-atherosclerotic properties, is associated with the presence of atherosclerosis and reduces plaque formation in experimental animals. Objectives: The present study aimed to investigate the chronic effect of BaP injection on hepatic FGF21 expression, as an anti-atherosclerotic hormone, in mice fed with or without an atherogenic diet (AtD). Methods: Eighteen C57BL/6J male mice (6 weeks) were randomly divided into six groups based on the dosage and diet. Blood samples were collected, and serum cholesterol, triglyceride, HDL-C, LDL-C, and glucose levels were measured. FGF21 expression was assessed by quantitative real-time PCR. Atherosclerotic lesions in mice were studied with Oil Red O (ORO) staining. Results: Benzo(a)pyrene causes a significant increase in liver FGF21 expression in a dose-dependent manner, and BaP co-exposure with AtD leads to a further increase in FGF21 expression. Additionally, the addition of BaP to AtD significantly increased the serum glucose, cholesterol, and LDL-C levels and accelerated the formation of atherosclerotic lesions. Besides, our findings showed that there is a significant positive correlation between FGF21 expression and glucose, cholesterol, LDL-C, and ORO-positive areas. Conclusions: Our findings revealed that BaP increases the expression of endogenous FGF21 in treated animals as a compensatory response to protect the heart from atherosclerosis induced by BaP and AtD.

Vaporized nicotine in utero results in reduced birthweight, increased locomotion, and decreased voluntary exercise, dependent on sex and diet in offspring.

Rationale Clinical research has shown that prenatal exposure to nicotine may result in increased obesity risk later in life. Preclinical research has corroborated this finding, but few studies have investigated inhaled nicotine or the interaction with diet on obesity risk. Objective The aim of this study was to investigate the effects of prenatal nicotine exposure on both direct and indirect obesity measures, with both sex and diet as factors. Methods Pregnant rats were exposed to either vehicle or nicotine vapor (24mg/mL or 59mg/mL) throughout the entire gestational period. Offspring from each treatment group were given either a normal diet or a high fat diet starting at postnatal day 22. Caloric intake, body weight, spontaneous locomotion, sleep/wake activity, and voluntary exercise were measured throughout adolescence. Pregnancy weight gain and pup birthweights were collected to further measure developmental effects of prenatal nicotine exposure. Results Both maternal weight gain during pregnancy and pup weight at birth were decreased with prenatal nicotine exposure. Early adolescent males showed increased spontaneous activity in the open field following prenatal nicotine exposure compared to vehicle counterparts, particularly those given high-fat diet. Additionally, high dose nicotine prenatal treated males ran significantly less distance on the running wheel in late adolescence compared to vehicle counterparts, in the normal diet group only. Conclusion The results presented here show decreased birthweight, hyperactivity, and decreased voluntary exercise in adolescence following prenatal nicotine exposure in dose, sex, and diet dependent manners, which could lead to increased obesity risk in adulthood.

Germination Indices and Seedling Growth Parameters in Maize under Salinity Stress with Varying Concentrations of Giberellic Acid (GA)

Salinity stress is one of the factors limiting seed emergence and seedling development in sunflower at New halfa scheme in East of Sudan. A study of two factors pots experiment was conducted during winter season of 2019 to evaluate germination indices and seedling growth characters of maize seeds under salinity stress with varying concentrations of synthetic growth regulator namely (GA3). The experiment was laid out according RCBD with three replications. Treatments consisted of five levels of salinity (0, 0.5, 1.0, 1.5 and 2% ) designated as Na0, Na0.5, Na1.0 ,Na1.5 and Na2.0, and four levels of GA3 were (0, 50,75 and 100 p.pm) designated as G0, G50, G75 and G100, respectively,. The studied parameters include germination indices; daily germination (MDG), daily germination speed (DGS), final germination percentage (FGP), mean emergence time (MET), mean Energy of germination (EE), emergence index (EI), coefficient of uniformity of emergence (CUE) and vigour index (SVI). Also, seedling fresh and dry weight, shoot length and root length were measured, and the shoot/root ratio was calculated. The negative effects of salinity stress significantly decreased germination indices and seedlings growth characters in a dose dependent manner. The results showed that, application of GA3 at medium levels concentration (specify those doses) increased all studied parameters. In conclusion, application of high Ga3 levels enhanced all germination indices and seedlings growth characters while high salinity levels reduced these traits of maize seeds. Such result has made the base for further study that should be conducted in different seasons with different growth regulators involving different parameters of other crops.

Enhancing Presurgical Infant Orthopedic Appliances: Characterization, Mechanics, and Biofilm Inhibition of a Novel Chlorhexidine-Halloysite Nanotube-Modified PMMA

Objectives. This in vitro study aimed to develop a novel nanocomposite acrylic resin with inherent antimicrobial properties. This study evaluated its effectiveness against microbial biofilm formation, while also assessing its physical and mechanical properties. Methods. Polymethylmethacrylate (PMMA) was modified with four different concentrations of chlorhexidine halloysite nanotubes (CHX-HNTs): 1%, 1.5%, 3%, and 4.5 wt.% by weight, along with a control group (0 wt.% CHX-HNTs). The biofilm inhibition ability of the modified CHX-HNTs acrylic against Candida albicans, Staphylococcus aureus, Streptococcus pneumoniae, and Streptococcus agalactiae was assessed using microtiter biofilm test. In addition, ten samples from each group were then tested for flexural strength, surface roughness, and hardness. Statistical analysis was performed using one-way ANOVA and Tukey’s test for comparison (P&lt;0.05). Results. CHX-HNTs effectively reduced the adhesion of Candida albicans and bacteria to the PMMA in a dose-dependent manner. The higher the concentration of CHX-HNTs, the greater the reduction in microbial adhesion, with the highest concentration (4.5 wt.%) showing the most significant effect with inhibition rates ≥98%. The addition of CHX-HNTs at any tested concentration (1%, 1.5%, 3%, and 4.5 wt.%) did not cause any statistically significant difference in the flexural strength, surface roughness, or hardness of the PMMA compared to the control group. Conclusions. The novel integration of CHX-HNT fillers shows promising results as an effective biofilm inhibitor on acrylic appliances. This new approach has the potential to successfully control infectious diseases without negatively affecting the mechanical properties of the acrylic resin. Clinical Relevance. The integration of CHX-HNTs into presurgical infant orthopedic appliances should be thoroughly assessed as a promising preventive measure to mitigate microbial infections. This evaluation holds significant potential for controlling infectious diseases among infants with cleft lip and palate, thereby offering a valuable contribution to their overall well-being.

Effect of Propolis on Salivary Glands After Radiotherapy.

To investigate the effects of increasing propolis doses on salivary glands exposed to radiotherapy (RT). Thirty-seven rats were divided into 4 groups: The control group (G0, n: 7), G1 group (n: 10), G2 group (n: 10), and G3 group (n: 10). The rats in the G1 group received 15 Gray (Gy) RT only to the head and neck area. The rats in the G2 and G3 groups received 15 Gy RT for the head and neck area along with 100mg/kg/ml and 200mg/kg/ml of propolis. The parotid, submandibular, and sublingual glands of rats were immunohistochemically stained with aquaporin-1 (AQP-1) and aquaporin-5 (AQP-5). They were also evaluated for malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPO), total antioxidant (TAS), and total oxidant status (TOS). AQP-1 and AQP-5 values were highest in G0 group followed by G3, G2, and G1 groups in decreasing order. The MDA and TOS values were highest in G1 group, which was followed by G2, G3, and G0 groups. The highest GPO, SOD, and TAS values were observed in G0 group followed by G3, G2, and G1 groups in decreasing order. It was found that propolis increased antioxidant products and decreased oxidative products in the salivary glands receiving RT in parallel with the dose increase. Similarly, in the groups receiving propolis, an increase in the immune expression of aquaporin molecules was detected in a dose-dependent manner. Based on these data, it could be stated that propolis has a healing effect on the salivary glands exposed to RT.

Novel Functional Food Properties of Forest Onion (Eleutherine bulbosa Merr.) Phytochemicals for Treating Metabolic Syndrome: New Insights from a Combined Computational and In Vitro Approach

Metabolic syndrome is a global health problem. The use of functional foods as dietary components has been increasing. One food of interest is forest onion extract (FOE). This study aimed to investigate the effect of FOE on lipid and glucose metabolism in silico and in vitro using the 3T3-L1 mouse cell line. This was a comprehensive study that used a multi-modal computational network pharmacology analysis and molecular docking in silico and 3T3-L1 mouse cells in vitro. The phytochemical components of FOE were analyzed using untargeted ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS). Next, an in silico analysis was performed to determine FOE’s bioactive compounds, and a toxicity analysis, protein target identification, network pharmacology, and molecular docking were carried out. FOE’s effect on pancreatic lipase, α-glucosidase, and α-amylase inhibition was determined. Finally, we determined its effect on lipid accumulation and MAPK8, PPARG, HMGCR, CPT-1, and GLP1 expression in the preadipocyte 3T3-L1 mouse cell line. We showed that the potential metabolites targeted glucose and lipid metabolism in silico and that FOE inhibited pancreatic lipase levels, α-glucosidase, and α-amylase in vitro. Furthermore, FOE significantly (p &lt; 0.05) inhibits targeted protein expressions of MAPK8, PPARG, HMGCR, CPT-1, and GLP-1 in vitro in 3T3-L1 mouse cells in a dose-dependent manner. FOE contains several metabolites that reduce pancreatic lipase levels, α-glucosidase, α-amylase, and targeted proteins associated with lipid and glucose metabolism in vitro.

Evaluation of propolis hepatotoxicity in male rats

Background: Propolis is a natural resinous combination that honeybees produce from the materials they gather from plant parts, buds, and exudates. Numerous favorable pharmacological qualities have been demonstrated for propolis. Objectives: This study aimed to investigate the hepatotoxic effects of ethanolic propolis extract on the liver of male rats. Methods: In this research, 40 male rats were divided randomly into five groups: 1. Control 2. Sham (solvent), and 3. Three experimental groups (ethanolic propolis extract at doses of 50,100 and 200 mg/kg). All materials were administered by oral gavage once daily for 13 consecutive days. On the 14th day, blood sampling was performed to measure serum levels of liver function enzymes and triglycerides. After deep induction of anesthesia, the liver of the rats was removed for histopathological studies. Data were analyzed using ANOVA and Tukey’s test (p &lt;0.05). Results: The data showed that the administration of propolis significantly increased the serum levels of aminotransferases in a dose-dependent manner and decreased triglycerides, accompanied by pathological changes. Conclusions: The results of this study indicate that the administration of propolis is associated with liver toxicity, and it seems that it should be consumed more carefully. Keywords: Alanine aminotransferase, Aspartate aminotransferase, Propolis, Hepatotoxicity Triglycerides

Long-Term Maintenance of Viable Human Endometrial Epithelial Cells to Analyze Estrogen and Progestin Effects

There are fewer investigations conducted on human primary endometrial epithelial cells (HPEECs) compared to human primary endometrial stromal cells (HPESCs). One of the main reasons is the scarcity of protocols enabling prolonged epithelial cell culture. Even though it is possible to culture HPEECs in 3D over a longer period of time, it is technically demanding. In this study, we successfully established a highly pure, stable, and long-term viable human conditionally reprogrammed endometrial epithelial cell line, designated as eCRC560. These cells stained positive for epithelial markers, estrogen and progesterone receptors, and epithelial cell–cell contacts but negative for stromal and endothelial cell markers. Estradiol (ES) reduced the abundance of ZO-1 in a time- and dose-dependent manner, in contrast to the dose-dependent increase with the progestin dienogest (DNG) when co-cultured with HPESCs. Moreover, ES significantly increased cell viability, cell migration, and invasion of the eCRC560 cells; all these effects were inhibited by pretreatment with DNG. DNG withdrawal led to a significantly disrupted monolayer of eCRC560 cells in co-culture with HPESCs, yet it markedly increased the adhesion of eCRC560 to the human mesothelial MeT-5A cells. The long-term viable eCRC560 cells are suitable for in vitro analysis of HPEECs to study the epithelial compartment of the human endometrium and endometrial pathologies.

Fish novel TRIM protein FTR14 negatively regulates interferon response by targeting TBK1-IRF3

Growing evidence highlights that tripartite motif (TRIM) family members exert crucial functions in host defense upon infection with microbial pathogens. Of note, the roles of a subfamily of fish-specific TRIM (FTR, also named as finTRIM) which have no true orthologues in mammals were largely unexplored. In this study, a novel FTR gene from grouper (EcFTR14) was identified, and its effects on fish virus infection and host interferon response were investigated. EcFTR14 contained RING, B-Box, and PRY-SPRY domains, and shared 54.81% identity to zebrafish FTR14. The transcription levels of EcFTR14 were markedly induced upon fish virus infection. EcFTR14 was mainly localized in the cytoplasm as small spots or bright aggregates. The overexpression of EcFTR14 could markedly enhance the replication of RGNNV or SGIV. Moreover, overexpression of EcFTR14 reduced the mRNA level of IFN-associated immune signaling molecules, suggesting that the antiviral action of EcFTR14 was due to the negative regulation on IFN response. EcFTR14 was found to interact with EcTBK1, EcIRF3 and EcIRF7, but not with EcSTING. Overexpression of EcFTR14 markedly suppressed the transcription levels of IFN-signaling related molecules evoked by EcSTING and EcTBK1. In addition, overexpression of EcFTR14 could degrade EcTBK1, EcIRF3 and EcIRF7 in a dose-dependent manner, that resulted in the suppression of antiviral effects induced by EcIRF3 and EcTBK1. Collectively, our findings illuminated that EcFTR14, a novel member of FTR family, acted as a pro-viral factor through negatively regulating targeting TBK1-IRF3 activated antviral action. The present study will for the first time shed light on the regulatory roles of FTR14 in innate immune response upon fish virus infection.

Effect of Neurotropic and Immunotropic Drugs on Leukocyte Elastase Activity In Vitro.

Leukocyte elastase is a marker of inflammation. Previously, a relationship was found between the severity of mental disorders in patients and elastase-like activity of blood plasma. The effect of various neurotropic drugs on leukocyte elastase activity was analyzed in an in vitro experiment. We revealed an inhibitory effect of the benzodiazepine tranquilizers diazepam and bromodihydrochlorophenylbenzodiazepine and immunomodulators aminodihydrophthalazinedione and diclofenac on the plasma elastase-like activity of healthy donors and pure human neutrophil elastase. The antipsychotics chlorpromazine and alimemazine, as well as the nootropic vinpocetine increased elastase-like activity in a dose-dependent manner. The activating effect of chlorpromazine and vinpocetine, but not alimemazine, was reproduced in neutrophil elastase. We hypothesized that these drugs can affect the development of inflammatory reactions in the complex therapy of mental disorders.

Molybdenum-Induced Oxidative Stress and Histopathological Alterations in the Epididymis

Molybednum is one of the trace elements required for proper functioning of the human body. The present study was designed to assess the effects of molybdenum on accessory reproductive organ i.e. epididymis. Male Wistar rats were administered with three different doses of molybdenum (50, 100 and 150 mg/kg body weight) for 60 days. A recovery study was also conducted in the highest dose group for which half the animals of the group were left untreated for the next 60 days. Exposure to molybdenum induced a significant decline in superoxide dismutase, ascorbic acid and glutathione while lipid peroxidation revealed a significant increase in epididymis in dose-dependent manner. Significant reduction in epididymal epithelial cells height, short sterocilia, increased intertubular stroma, decreased epididymal sperm reserve in the lumen and reduction in the diameter of cauda epididymal tubules with a relatively large amount of intertubular stroma was observed in epididymis of mice treated with molybdenum which might be correlated with enhanced oxidative stress. In the recovery group, significant improvement in antioxidant parameters and histoarchitecture of epididymis was observed indicating toxic effects of molybdenum can be reversed by cessation of exposure.

Dietary glycine supplementation enhances syntheses of creatine and glutathione by tissues of hybrid striped bass (Morone saxatilis ♀ × Morone chrysops ♂) fed soybean meal-based diets

BackgroundWe recently reported that supplementing glycine to soybean meal-based diets is necessary for the optimum growth of 5- to 40-g (Phase-I) and 110- to 240-g (Phase-II) hybrid striped bass (HSB), as well as their intestinal health. Although glycine serves as an essential substrate for syntheses of creatine and glutathione (GSH) in mammals (e.g., pigs), little is known about these metabolic pathways or their nutritional regulation in fish. This study tested the hypothesis that glycine supplementation enhances the activities of creatine- and GSH-forming enzymes as well as creatine and GSH availabilities in tissues of hybrid striped bass (HSB; Morone saxatilis♀ × Morone chrysops♂).MethodsPhase-I and Phase-II HSB were fed a soybean meal-based diet supplemented with 0%, 1%, or 2% glycine for 8 weeks. At the end of the 56-d feeding, tissues (liver, intestine, skeletal muscle, kidneys, and pancreas) were collected for biochemical analyses.ResultsIn contrast to terrestrial mammals and birds, creatine synthesis occurred primarily in skeletal muscle from all HSB. The liver was most active in GSH synthesis among the HSB tissues studied. In Phase-I HSB, supplementation with 1% or 2% glycine increased (P < 0.05) concentrations of intramuscular creatine (15%–19%) and hepatic GSH (8%–11%), while reducing (P < 0.05) hepatic GSH sulfide (GSSG)/GSH ratios by 14%–15%, compared with the 0-glycine group; there were no differences (P > 0.05) in these variables between the 1% and 2% glycine groups. In Phase-II HSB, supplementation with 1% and 2% glycine increased (P < 0.05) concentrations of creatine and GSH in the muscle (15%–27%) and liver (11%–20%) in a dose-dependent manner, with reduced ratios of hepatic GSSG/GSH in the 1% or 2% glycine group. In all HSB, supplementation with 1% and 2% glycine dose-dependently increased (P < 0.05) activities of intramuscular arginine:glycine amidinotransferase (22%–41%) and hepatic γ-glutamylcysteine synthetase (17%–37%), with elevated activities of intramuscular guanidinoacetate methyltransferase and hepatic GSH synthetase and GSH reductase in the 1% or 2% glycine group. Glycine supplementation also increased (P < 0.05) concentrations of creatine and activities of its synthetic enzymes in tail kidneys and pancreas, and concentrations of GSH and activities of its synthetic enzymes in the proximal intestine.ConclusionsSkeletal muscle and liver are the major organs for creatine and GSH syntheses in HSB, respectively. Dietary glycine intake regulates creatine and GSH syntheses by both Phase-I and Phase-II HSB in a tissue-specific manner. Based on the metabolic data, glycine is a conditionally essential amino acid for the growing fish.