Differential Action Research Articles

Atorvastatin favorably modulates a clinical hepatocellular carcinoma risk gene signature.

Lipophilic but not hydrophilic statins have been shown to be associated with reduced risk for hepatocellular carcinoma (HCC) in patients with chronic viral hepatitis. We investigated differential actions of lipophilic and hydrophilic statins and their ability to modulate a clinical prognostic liver signature (PLS) predicting HCC risk in patients with liver disease. Hepatitis C virus (HCV)–infected Huh7.5.1 cells, recently developed as a model to screen HCC chemopreventive agents, were treated with lipophilic statins (atorvastatin and simvastatin) and hydrophilic statins (rosuvastatin and pravastatin), and then analyzed by RNA sequencing and PLS. Lipophilic statins, particularly atorvastatin, more significantly suppressed the HCV‐induced high‐risk pattern of PLS and genes in YAP and AKT pathway implicated in fibrogenesis and carcinogenesis, compared with the hydrophilic statins. While atorvastatin inhibited YAP activation through the mevalonate pathway, the distinctive AKT inhibition of atorvastatin was mediated by stabilizing truncated retinoid X receptor alpha, which has been known to enhance AKT activation, representing a target for HCC chemoprevention. In addition, atorvastatin modulated the high‐risk PLS in an in vitro model of nonalcoholic fatty liver disease (NAFLD). Conclusion: Atorvastatin distinctively inhibits YAP and AKT activation, which are biologically implicated in HCC development, and attenuates a high‐risk PLS in an in vitro model of HCV infection and NAFLD. These findings suggest that atorvastatin is the most potent statin to reduce HCC risk in patients with viral and metabolic liver diseases.

Bisphenol S Alters the Steroidome in the Preovulatory Follicle, Oviduct Fluid and Plasma in Ewes With Contrasted Metabolic Status.

Bisphenol A (BPA), a plasticizer and endocrine disruptor, has been substituted by bisphenol S (BPS), a structural analogue that had already shown adverse effects on granulosa cell steroidogenesis. The objective of this study was to assess the effect of chronic exposure to BPS, a possible endocrine disruptor, on steroid hormones in the ovary, oviduct and plasma using the ewe as a model. Given the interaction between steroidogenesis and the metabolic status, the BPS effect was tested according to two diet groups. Eighty adult ewes were allotted to restricted (R) and well-fed (WF) groups, that were further subdivided into two subgroups. Ewes were exposed to 50 µg BPS/kg/day in their diet (R50 and WF50 groups) or were unexposed controls (R0 and WF0 groups). After at least 3 months of BPS exposure, preovulatory follicular fluid, oviduct fluid and plasma were collected and steroid hormones were analyzed by gas chromatography coupled with tandem mass spectrometry (GC-MS/MS). A deleterious effect of restricted diet on the volume of oviduct fluid and numbers of pre-ovulatory follicles was observed. Exposure to BPS impaired estradiol concentrations in both follicular and oviduct fluids of well-fed ewes and progesterone, estradiol and estrone concentrations in plasma of restricted ewes. In addition, a significant interaction between metabolic status and BPS exposure was observed for seven steroids, including estradiol. In conclusion, BPS acts in ewes as an endocrine disruptor with differential actions according to metabolic status.

Effects of Acute Triiodothyronine Treatment in Patients with Anterior Myocardial Infarction Undergoing Primary Angioplasty: Evidence from a Pilot Randomized Clinical Trial (ThyRepair Study).

Background: Thyroid hormone has a differential action on healthy and ischemic heart. Triiodothyronine (T3) administration improved postischemic cardiac function while it limited apoptosis in experimentally induced ischemia. Thus, the present study investigated the potential effects of acute liothyronine (LT3) treatment in patients with anterior myocardial infarction. Methods: This study is a pilot, randomized, double-blind, placebo-controlled trial (ThyRepair study). We randomized 52 patients and analyzed data from 37 patients (n = 16 placebo and n = 21 LT3), per prespecified per protocol analysis. We excluded three patients who had died of cardiovascular causes (one in placebo and two in LT3 arm), four with small infarct size below a pre-specified threshold (in the placebo arm), and the rest, who lacked follow-up data. LT3 treatment started after stenting as an intravenous (i.v.) bolus injection of 0.8 μg/kg of LT3 followed by a constant infusion of 0.113 μg/kg/h i.v. for 48 hours. All patients had cardiac magnetic resonance (CMR) at hospital discharge and 6 months follow-up. The primary end point was CMR left ventricular (LV) ejection fraction (LVEF) and secondary endpoints were LV volumes, infarct volume (IV), and safety. Results: The CMR LVEF% at 6 months was 53.6 ± 9.5 for the LT3-treated group and 48.6 ± 11 for placebo, p = 0.15. Acute LT3 treatment resulted in a significantly lower LV end-diastolic volume index (92.2 ± 16.8 mL/m2 vs. 107.5 ± 22.2, p = 0.022) and LV systolic volume index (47.5 ± 13.9 mL/m2 vs. 61.3 ± 21.7, p = 0.024) at hospital discharge, but not at 6 months. There was no statistically significant difference in CMR IV at hospital discharge between the groups (p = 0.24). CMR IV tended to be lower in the LT3-treated group at 6 months (18.7 ± 9.5 vs. 25.9 ± 11.7, in placebo, p = 0.05). Serious, life-threatening events related to LT3 treatment were not observed. A tendency for an increased incidence of atrial fibrillation (AF) was found in the LT3 group during the first 48 hours (19% for T3 group vs. 5% for placebo, p = 0.13). Conclusion: This pilot randomized, placebo-controlled trial study suggests potential favorable effects (acute cardiac dilatation and 6-month IV) as well as potential concerns regarding a higher risk of AF after LT3 administration early after myocardial infarction, which should be tested in a larger scale study.

TSLP and IL-33 distinctively modulate the allergic response by differentially modulating Th2 cells and ILC2s

Abstract Alarmins are endogenous chemotactic or immune-activating proteins released after cell injury or immune activation, and these include thymic stromal lymphopoietin (TSLP) and IL-33. Mice lacking these cytokines exhibit defective type 2 responses, but there is controversy regarding the roles of these cytokines in allergic responses in vivo. Here, we demonstrate that the nature of the allergic response is dictated by the alarmin. Mice lacking IL-33 were resistant to pulmonary inflammation induced by papain, but they exhibited normal OVA-induced allergic inflammation. In contrast, mice lacking either TSLP or TSLPR had significantly lower pulmonary inflammation in response to OVA challenge but responded normally to papain. Moreover, while intranasal papain challenge rapidly induced the release of IL-33, OVA challenge predominantly released TSLP. Steady-state pulmonary ILC2s were activated in response to IL-33, and correspondingly, Il33 KO mice showed a similar defect in effector Th2 generation to that observed in animals lacking ILC2s. In contrast to IL-33, TSLP did not activate steady-state ILC2s. Instead, conditional deletion of TSLPR in T cells hampered Th2 cell generation in the OVA-induced asthma model, but these mice responded normally to papain. These data demonstrate that Th2 cells rely on IL-33-stimulated ILC2s, whereas in the OVA-induced asthma model, TSLP acts on Th2 cells to promote type 2 responses. Thus, TSLP and IL-33 are both alarmins, but they have distinctive effects on allergic responses based on their different release kinetics and on their differential actions on Th2 cells versus ILC2s. These findings indicate approaches to selectively target different allergic responses, with potential translational impact. This research was supported by the Intramural Research Programs of NIAID and NHLBI, NIH.

Proarrhythmic risk assessment of drugs by dVm /dt shapes using the convolutional neural network.

Comprehensive in vitro Proarrhythmia Assay (CiPA) projects for assessing proarrhythmic drugs suggested a logistic regression model using qNet as the Torsades de Pointes (TdP) risk assessment biomarker, obtained from in silico simulation. However, using a single in silico feature, such as qNet, cannot reflect whole characteristics related to TdP in the entire action potential (AP) shape. Thus, this study proposed a deep convolutional neural network (CNN) model using differential action potential shapes to classify three proarrhythmic risk levels: high, intermediate, and low, considering both characteristics related to TdP not only in the depolarization phase but also the repolarization phase of AP shape. We performed an in silico simulation and got AP shapes with drug effects using half‐maximal inhibitory concentration and Hill coefficients of 28 drugs released by CiPA groups. Then, we trained the deep CNN model with the differential AP shapes of 12 drugs and tested it with those of 16 drugs. Our model had a better performance for classifying the proarrhythmic risk of drugs than the traditional logistic regression model using qNet. The classification accuracy was 98% for high‐risk level drugs, 94% for intermediate‐risk level drugs, and 89% for low‐risk level drugs.

A mass spectrometry-based non-radioactive differential radial capillary action of ligand assay (DRaCALA) to assess ligand binding to proteins.

Binding of ligands to macromolecules changes their physicochemical and enzymatic characteristics. Cyclic di‐GMP is a second messenger involved in motility/sessility and acute/chronic infection life style transition. Although the GGDEF domain, predominantly a diguanylate cyclase, represents one of the most abundant bacterial domain superfamilies, the number of cyclic di‐GMP receptors falls short. To facilitate screening for cyclic di‐nucleotide binding proteins, we describe a non‐radioactive, matrix‐assisted laser desorption and ionization time‐of‐flight (MALDI‐TOF)‐based modification of the widely applied differential radial capillary action of ligand assay (DRaCALA). The results of this assay suggest that the diguanylate cyclase/phosphodiesterase variant YciRFec101, but not selected catalytic mutants, bind cyclic di‐GMP.Highlights Cyclic di‐nucleotides are ubiquitous second messengers in bacteria. However, few receptors have been identified.Previous screening of cell lysates by differential radial capillary action of ligand assay (DRaCALA) using radioactive ligand identified cyclic di‐nucleotide binding proteins.A MALDI‐TOF‐based DRaCALA was developed to detect cyclic di‐nucleotide binding as a non‐radioactive alternative.Known cyclic di‐GMP binding proteins were verified and potential cyclic di‐GMP binding proteins were identified.

Striatal Neuronal Ensembles Reveal Differential Actions of Amantadine and Clozapine to Ameliorate Mice L-DOPA-Induced Dyskinesia

Amantadine and clozapine have proved to reduce abnormal involuntary movements (AIMs) in preclinical and clinical studies of L-DOPA-Induced Dyskinesias (LID). Even though both drugs decrease AIMs, they may have different action mechanisms by using different receptors and signaling profiles. Here we asked whether there are differences in how they modulate neuronal activity of multiple striatal neurons within the striatal microcircuit at histological level during the dose-peak of L-DOPA in ex-vivo brain slices obtained from dyskinetic mice. To answer this question, we used calcium imaging to record the activity of dozens of neurons of the dorsolateral striatum before and after drugs administration in vitro. We also developed an analysis framework to extract encoding insights from calcium imaging data by quantifying neuronal activity, identifying neuronal ensembles by linking neurons that coactivate using hierarchical cluster analysis and extracting network parameters using Graph Theory. The results show that while both drugs reduce LIDs scores behaviorally in a similar way, they have several different and specific actions on modulating the dyskinetic striatal microcircuit. The extracted features were highly accurate in separating amantadine and clozapine effects by means of principal components analysis (PCA) and support vector machine (SVM) algorithms. These results predict possible synergistic actions of amantadine and clozapine on the dyskinetic striatal microcircuit establishing a framework for a bioassay to test novel antidyskinetic drugs or treatments in vitro.

Ternary poly(2-ethyl-2-oxazoline)-polyvinylpyrrolidone-graphene nanocomposites: Thermal, electrical, dielectric, mechanical, and antibacterial profiling

Graphene-based poly(2-ethyl-2-oxazoline) (PEOX) and polyvinylpyrrolidone (PVP) blend-matrix nanocomposites were prepared employing different weight percentages of graphene nanoplatelets as filler by ultrasonication assisted solution casting method. These nanocomposites were explored for their thermal, electrical, dielectric, and mechanical properties, and antimicrobial efficiency. Thermogravimetric analysis demonstrated that graphene operates as a barrier to limit thermal diffusion across the PEOX-PVP blend matrix, and hence, improve thermal stability of nanocomposites. The dielectric and electric properties such as dielectric constant, dielectric loss, loss tangent and electrical conductivity of PEOX-PVP-10 wt% graphene nanocomposite were found to increase with temperature. The presence of semi-circles in the Cole-Cole plot indicated the existence of a relaxation process in the conduction mechanism of the nanocomposite. AC electrical conductivity, σAC, of PEOX-PVP-10 wt% graphene nanocomposite was found to obey Jonscher's power law. The temperature-dependent behavior of frequency exponent, s, of σAC discusses the applicability of correlated barrier hopping (CBH) model. The extracted DC conductivity from AC conductivity studies was found to be temperature-dependent and obey Arrhenius relation with activation energy of conduction, Ea, of 0.41 eV and 0.39 eV in the lower and higher temperature regions, respectively. The mechanical properties of nanocomposites were enhanced dramatically when graphene loading was increased, demonstrating that a better interaction exists between graphene and the PEOX-PVP blend matrix. PEOX-PVP-15 wt% graphene nanocomposite showed superior mechanical properties (tensile strength: 9.18 MPa and Young's modulus: 3.19 MPa) among the synthesized nanocomposites. Further, the antibacterial activities of these nanocomposites against Gram-negative (E. coli) and Gram-positive (E. facecalis) bacteria revealed differential action.

Sleep fragmentation engages stress-responsive circuitry, enhances inflammation and compromises hippocampal function following traumatic brain injury

Traumatic brain injury (TBI) impairs the ability to restore homeostasis in response to stress, indicating hypothalamic-pituitary-adrenal (HPA)-axis dysfunction. Many stressors result in sleep disturbances, thus mechanical sleep fragmentation (SF) provides a physiologically relevant approach to study the effects of stress after injury. We hypothesize SF stress engages the dysregulated HPA-axis after TBI to exacerbate post-injury neuroinflammation and compromise recovery. To test this, male and female mice were given moderate lateral fluid percussion TBI or sham-injury and left undisturbed or exposed to daily, transient SF for 7- or 30-days post-injury (DPI). Post-TBI SF increases cortical expression of interferon- and stress-associated genes characterized by inhibition of the upstream regulator NR3C1 that encodes glucocorticoid receptor (GR). Moreover, post-TBI SF increases neuronal activity in the hippocampus, a key intersection of the stress-immune axes. By 30 DPI, TBI SF enhances cortical microgliosis and increases expression of pro-inflammatory glial signaling genes characterized by persistent inhibition of the NR3C1 upstream regulator. Within the hippocampus, post-TBI SF exaggerates microgliosis and decreases CA1 neuronal activity. Downstream of the hippocampus, post-injury SF suppresses neuronal activity in the hypothalamic paraventricular nucleus indicating decreased HPA-axis reactivity. Direct application of GR agonist, dexamethasone, to the CA1 at 30 DPI increases GR activity in TBI animals, but not sham animals, indicating differential GR-mediated hippocampal action. Electrophysiological assessment revealed TBI and SF induces deficits in Schaffer collateral long-term potentiation associated with impaired acquisition of trace fear conditioning, reflecting dorsal hippocampal-dependent cognitive deficits. Together these data demonstrate that post-injury SF engages the dysfunctional post-injury HPA-axis, enhances inflammation, and compromises hippocampal function. Therefore, external stressors that disrupt sleep have an integral role in mediating outcome after brain injury.

A study of gonadal dysfunction in women with thyroid disease

Background: Thyroid diseases are among the commonest endocrine disorders. It effects growth, embryonic development, tissue differentiation, maturation, metabolism, increases chronotropic and inotropic action of heart. The aim was to study gonadal dysfunction in women with thyroid disease; to study levels of FSH, LH, prolactin, estrogen and testosterone in hypothyroid and hyperthyroid.Methods: All participants fulfilling the inclusion criteria were interviewed as per proforma and medical details were recorded in proforma sheet and patients were subjected to necessary blood investigations.Results: Among 80 study subjects, majority were in the age group 26-35 years (60.1%) and majority had overt hypothyroid disease (72.5%). Symptoms of hypothyroid subjects mainly were fatigue, weight gain, hair loss, oligomenorrhea. Hyperthyroid subjects had fatigue, palpitations, oligomenorrhea, polymenorrhea. Prolactin had positive correlation with TSH in hypothyroid subjects, their LH, FSH levels were normal and sex steroid levels were low. In contrary, majority of the hyperthyroid subjects had normal prolactin and FSH, high LH and high sex steroid levels.Conclusions: The study demonstrated that thyroid disease in women causes menstrual abnormalities due to altered gonadotropin patterns which in turn alter ovarian hormone pattern. Hypothyroidism is associated with hyperprolactinemia leading to anovulation and abnormal menstrual cycle. Hyperthyroidism is associated with increased levels of LH, estrogen and testosterone with normal levels of PRL and FSH. Screening, identifying and correcting thyroid disease should be undertaken to prevent infertility as early as possible.

Specific Expression of KCC2 in the α Cells of Normal and Type 1 Diabetes Model Mouse Pancreatic Islets.

Gamma-aminobutyric acid (GABA) is an inhibitory neurotransmitter in the mature brain; however, it acts excitatory during development. This difference in action depends on the intracellular chloride ion concentration, primarily regulated by potassium chloride co-transporter2 (KCC2). Sufficient KCC2 expression results in its inhibitory action. GABA is also abundant in pancreatic islets, where it acts differentially on the islet cells, and is involved in carbohydrate metabolism. However, the mechanisms underlying the differential action remain unknown. We performed immunohistochemistry for glutamic acid decarboxylase (GAD), a synthetic enzyme for GABA, and KCC2 in normal adult islets. GAD was co-localized with insulin in β cells, whereas KCC2 was expressed in glucagon-positive α cells. These results are in line with previous observations that GABA decreases glucagon release but increases insulin release, and suggest that GABA and insulin may work together in reducing blood glucose levels under hyperglycemia. Next, we examined the streptozotocin-induced type1 diabetes mellitus mouse model. GAD and insulin expression levels were markedly decreased. KCC2 was expressed in glucagon-positive cells, whereas insulin- and somatostatin-positive cells were KCC2-negative. These findings suggest that in diabetes model, reduced GABA release may cause disinhibition of glucagon release, resulting in increased blood sugar levels and the maintenance of hyperglycemic state.

Neuroimaging Mechanism of Cognitive Behavioral Therapy in Pain Management.

Purpose. To review the recent neuroimaging studies on cognitive-behavioral therapy (CBT) for pain management, with the aim of exploring possible mechanisms of CBT. Recent Findings. Current studies can be divided into four categories, mixed pain, fibromyalgia, migraine, and experimental pain, based on the type of disease included, with the same or different changes of brain regions after CBT intervention. According to structural and functional MRI analyses, changes of brain gray matter volume, activation and deactivation of brain regions, and intrinsic connectivity between brain regions were observed after CBT sessions. The brain regions involved mainly included some areas related to cognitive and emotional regulation. After comparison, the DLPFC, OFC, VLPFC, PCC and amygdala were found to be recurrent in multiple studies and may be key regions for CBT intervention in pain management. In the treatment of mixed chronic pain, CBT may decrease the gray matter volume of DLPFC, reduce ICN connection of OFC within the DAN network, and increase fALFF of the PCC. For FM intervention, CBT may activate the bilateral OFC and VLPFC, while in migraine, only the right OFC, VLPFC, and DLPFC were found to be more activated after CBT. In addition, the differential action of the left and right amygdala has also been shown in the latest study of migraine. In heat-evoked pain, CBT may increase the deactivation of the PCC, the connectivity between the DMN and right VLPFC, while diminishing the deactivation of VLPFC. Summary. After CBT, the brain showed stronger top-down pain control, cognitive reassessment, and altered perception of stimulus signals (chronic pain and repeated acute pain). The DLPFC, OFC, VLPFC, PCC, and amygdala may be the key brain regions in CBT intervention of pain.

The context of the ribosome binding site in mRNAs defines specificity of action of kasugamycin, an inhibitor of translation initiation

Kasugamycin (KSG) is an aminoglycoside antibiotic widely used in agriculture and exhibits considerable medical potential. Previous studies suggested that KSG interferes with translation by blocking binding of canonical messenger RNA (mRNA) and initiator transfer tRNA (tRNA) to the small ribosomal subunit, thereby preventing initiation of protein synthesis. Here, by using genome-wide approaches, we show that KSG can interfere with translation even after the formation of the 70S initiation complex on mRNA, as the extent of KSG-mediated translation inhibition correlates with increased occupancy of start codons by 70S ribosomes. Even at saturating concentrations, KSG does not completely abolish translation, allowing for continuing expression of some Escherichia coli proteins. Differential action of KSG significantly depends on the nature of the mRNA residue immediately preceding the start codon, with guanine in this position being the most conducive to inhibition by the drug. In addition, the activity of KSG is attenuated by translational coupling as genes whose start codons overlap with the coding regions or the stop codons of the upstream cistrons tend to be less susceptible to drug-mediated inhibition. Altogether, our findings reveal KSG as an example of a small ribosomal subunit-targeting antibiotic with a well-pronounced context specificity of action.

Differential action of pro-angiogenic and anti-angiogenic components of Danhong injection in ischemic vascular disease or tumor models

ObjectiveWe investigate the chemical basis and mechanism of angiogenesis regulation by a multicomponent Chinese medicine Danhong injection (DHI).MethodsDHI was fractionated and screened for angiogenesis activities by in vitro tube formation and migration assays. The composition of DHI components was determined by UPLC. The effects of the main active monomers on angiogenesis-related gene and protein expression in endothelial cells were determined by qPCR and Western blotting analyses. Mouse hind limb ischemia and tumor implant models were used to verify the angiogenesis effects in vivo by Laser Doppler and bioluminescent imaging, respectively.ResultsTwo distinct chemical components, one promoting (pro-angiogenic, PAC) and the other inhibiting (anti-angiogenic, AAC) angiogenesis, were identified in DHI. PAC enhanced angiogenesis and improved recovery of ischemic limb perfusion while AAC reduced Lewis lung carcinoma growth in vivo in VEGFR-2-Luc mice. Among the PAC or AAC monomers, caffeic acid and rosmarinic acid upregulated TSP1 expression and downregulated KDR and PECAM expression. Caffeic acid and rosmarinic acid significantly decreased while protocatechuic aldehyde increased CXCR4 expression, which are consistent with their differential effects on EC migration.ConclusionsDHI is capable of bi-directional regulation of angiogenesis in disease-specific manner. The pro-angiogenesis activity of DHI promotes the repair of ischemic vascular injury, whereas the anti-angiogenesis activity inhibits tumor growth. The active pro- and anti-angiogenesis activities are composed of unique chemical combinations that differentially regulate angiogenesis-related gene networks.

Transient Receptor Potential Channels and Auditory Functions.

Significance: Transient receptor potential (TRP) channels are cation-gated channels that serve as detectors of various sensory modalities, such as pain, heat, cold, and taste. These channels are expressed in the inner ear, suggesting that they could also contribute to the perception of sound. This review provides more details on the different types of TRP channels that have been identified in the cochlea to date, focusing on their cochlear distribution, regulation, and potential contributions to auditory functions. Recent Advances: To date, the effect of TRP channels on normal cochlear physiology in mammals is still unclear. These channels contribute, to a limited extent, to normal cochlear physiology such as the hair cell mechanoelectrical transduction channel and strial functions. More detailed information on a number of these channels in the cochlea awaits future studies. Several laboratories focusing on TRPV1 channels have shown that they are responsive to cochlear stressors, such as ototoxic drugs and noise, and regulate cytoprotective and/or cell death pathways. TRPV1 expression in the cochlea is under control of oxidative stress (produced primarily by NOX3 NADPH oxidase) as well as STAT1 and STAT3 transcription factors, which differentially modulate inflammatory and apoptotic signals in the cochlea. Inhibition of oxidative stress or inflammation reduces the expression of TRPV1 channels and protects against cochlear damage and hearing loss. Critical Issues: TRPV1 channels are activated by both capsaicin and cisplatin, which produce differential effects on the inner ear. How these differential actions are produced is yet to be determined. It is clear that TRPV1 is an essential component of cisplatin ototoxicity as knockdown of these channels protects against hearing loss. In contrast, activation of TRPV1 by capsaicin protected against subsequent hearing loss induced by cisplatin. The cellular targets that are influenced by these two drugs to account for their differential profiles need to be fully elucidated. Furthermore, the potential involvement of different TRP channels present in the cochlea in regulating cisplatin ototoxicity needs to be determined. Future Directions: TRPV1 has been shown to mediate the entry of aminoglycosides into the hair cells. Thus, novel otoprotective strategies could involve designing drugs to inhibit entry of aminoglycosides and possibly other ototoxins into cochlear hair cells. TRP channels, including TRPV1, are expressed on circulating and resident immune cells. These receptors modulate immune cell functions. However, whether they are activated by cochlear stressors to initiate cochlear inflammation and ototoxicity needs to be determined. A better understanding of the function and regulation of these TRP channels in the cochlea could enable development of novel treatments for treating hearing loss. Antioxid. Redox Signal. 36, 1158-1170.

The Campylobacter jejuni Response Regulator and Cyclic-Di-GMP Binding CbrR Is a Novel Regulator of Flagellar Motility.

A leading cause of bacterial gastroenteritis, Campylobacter jejuni is also associated with broad sequelae, including extragastrointestinal conditions such as reactive arthritis and Guillain-Barré Syndrome (GBS). CbrR is a C. jejuni response regulator that is annotated as a diguanylate cyclase (DGC), an enzyme that catalyzes the synthesis of c-di-GMP, a universal bacterial second messenger, from GTP. In C. jejuni DRH212, we constructed an unmarked deletion mutant, cbrR−, and complemented mutant, cbrR+. Motility assays indicated a hyper-motile phenotype associated with cbrR−, whereas motility was deficient in cbrR+. The overexpression of CbrR in cbrR+ was accompanied by a reduction in expression of FlaA, the major flagellin. Biofilm assays and scanning electron microscopy demonstrated similarities between DRH212 and cbrR−; however, cbrR+ was unable to form significant biofilms. Transmission electron microscopy showed similar cell morphology between the three strains; however, cbrR+ cells lacked flagella. Differential radial capillary action of ligand assays (DRaCALA) showed that CbrR binds GTP and c-di-GMP. Liquid chromatography tandem mass spectrometry detected low levels of c-di-GMP in C. jejuni and in E. coli expressing CbrR. CbrR is therefore a negative regulator of FlaA expression and motility, a critical virulence factor in C. jejuni pathogenesis.

Gene action for yield and yield contributing traits in blackgram through generation mean analysis

Generation mean analysis was worked out for three crosses of blackgram viz., VBN(Bg) 4 x CO 5, VBN(Bg) 4 x APK 1 and VBN(Bg) 4 x Mash 1008. Five parameter model was utilized in this study to identify the gene actions of various yield and yield contributing traits. Most of the traits recorded the presence of epistasis gene action. The traits viz., pod length, the number of seeds per pod recorded additive-dominance model in the cross VBN(Bg) 4 x APK 1. Significant additive–dominance model for 100-seed weight recorded in the cross VBN(Bg) 4 x CO 5. Hence, these traits can be improved through pedigree breeding and simple selection. Significant additive or additive x additive gene action was observed for days to flowering, plant height, the number of pods per cluster and 100-seed weight in all crosses. The selection needs to be delayed due to the presence of additive or additive x additive gene action with an epistatic model. Other traits had differential gene action in each cross. Based on the results, it can be concluded that an appropriate selection programme needs to be followed for each cross based on the gene action. Key words: Blackgram, Generation mean analysis, Additive-dominance model, Epistatic model.

Estimation of genetic parameters and gene action among crosses of blackgram (Vigna mungo (L.) Hepper) for seed yield and its component traits

Generation mean analysis was employed among the crosses of MDU1 x TU68, VBN6 x TU68 and VBN8 x TU68 of blackgram to partition the mean into various genetic components viz., additive, dominance and epistasis. Five generations viz., P1, P2, F1, F2 and F3 obtained from the above three cross combinations were evaluated. The results of the scaling test showed that number of branches per plant in MDU 1 x TU 68, number of branches per plant and number of pods per cluster in VBN 6 x TU 68 and days to flowering and pod length in VBN 8 x TU 68 recorded additive gene action. Hence, these traits can be improved through pedigree breeding and simple selection. All other traits showed non additive model as one or both scales of C and D had significance. Days to flowering, plant height, number of branches per plant, number of clusters per plant, pod length, number of seeds per pod, 100-seed weight and single plant yield had additive gene action. In the case of epistasis, days to flowering had additive x additive type gene action in MDU 1 x TU68 and VBN6 x TU68 crosses, while the cross VBN8 x TU68 had additive gene action alone. Hence, the selection at later generations is effective to improve these traits. Other traits had differential gene action in each cross. Based on the results, it can be concluded that an appropriate selection programme needs to be devised for each cross based on the gene action. Key words: Blackgram, Generation mean analysis, Epistasis, Five parameters, Seed Yield.

Differential actions of nanoparticles and nanoemulsion synthesized from Colletotrichum siamense on food borne pathogen

Food spoilage and spread of diseases through food has been an increasing problem. Nanoparticles are being used in various applications, and one of its applications is its use in effective food packaging, preservation, and storage. In this study, endophytic fungal extract from Colletotrichum siamense was used as base material for the synthesis of silver nanoparticles and the blend of endophytic fungal extract and tea tree oil was utilized for formulating silver based nanoemulsion. Both were characterized by UV–Vis spectroscopy, FTIR, Scanning electron microscopy (SEM). The efficacy of nanoparticles and nanoemulsion against food borne pathogens were validated by using antibacterial assays. From the biofilm assay it was observed that both nanoparticles and naoemulsion controlled the biofilm formation in both the strains Enterobacter kobei SFV 1 and Klebsiella oxytoca SFV 3, however nanoemulsion showed better biofilm inhibitory effect than the nanoparticles. The results showed that the green silver nanoparticles and tea tree oil mediated nanoemulsion obtained from the endophytic fungal extract, could be used to develop into product that can be used in storage of food products to protect from food borne pathogens.

Differential Actions of Muscarinic Receptor Subtypes in Gastric, Pancreatic, and Colon Cancer.

Cancers arising from gastrointestinal epithelial cells are common, aggressive, and difficult to treat. Progress in this area resulted from recognizing that the biological behavior of these cancers is highly dependent on bioactive molecules released by neurocrine, paracrine, and autocrine mechanisms within the tumor microenvironment. For many decades after its discovery as a neurotransmitter, acetylcholine was thought to be synthesized and released uniquely from neurons and considered the sole physiological ligand for muscarinic receptor subtypes, which were believed to have similar or redundant actions. In the intervening years, we learned this former dogma is not tenable. (1) Acetylcholine is not produced and released only by neurons. The cellular machinery required to synthesize and release acetylcholine is present in immune, cancer, and other cells, as well as in lower organisms (e.g., bacteria) that inhabit the gut. (2) Acetylcholine is not the sole physiological activator of muscarinic receptors. For example, selected bile acids can modulate muscarinic receptor function. (3) Muscarinic receptor subtypes anticipated to have overlapping functions based on similar G protein coupling and downstream signaling may have unexpectedly diverse actions. Here, we review the relevant research findings supporting these conclusions and discuss how the complexity of muscarinic receptor biology impacts health and disease, focusing on their role in the initiation and progression of gastric, pancreatic, and colon cancers.