Short periods of bipolar anodal TDCS induce no instantaneous dose-dependent increase in cerebral blood flow in the targeted human motor cortex

Background: Anodal transcranial direct current stimulation (aTDCS) of primary motor hand area (M1-HAND) can enhance corticomotor excitability. Yet, it is still unknown which current intensity produces the strongest effect on regional neural activity. Magnetic resonance imaging (MRI) combined with pseudo-continuous Arterial Spin Labeling (pc-ASL MRI) can map regional cortical blood flow (rCBF) and may thus be useful to probe the relationship between current intensity and neural response at the individual level. Objective: Here we employed pc-ASL MRI to map acute rCBF changes during short-duration aTDCS of left M1-HAND. Using the rCBF response as a proxy for regional neuronal activity, we investigated if short-duration aTDCS produces an instantaneous dose-dependent rCBF increase in the targeted M1-HAND that may be useful for individual dosing. Methods: Nine healthy right-handed participants received 30 seconds of aTDCS at 0.5, 1.0, 1.5, and 2.0 mA with the anode placed over left M1-HAND and cathode over the right supraorbital region. Concurrent pc-ASL MRI at 3 T probed TDCS-related rCBF changes in the targeted M1-HAND. Movement-induced rCBF changes were also assessed. Results: Apart from a subtle increase in rCBF at 0.5 mA, short-duration aTDCS did not modulate rCBF in the M1-HAND relative to no-stimulation periods. None of the participants showed a dose-dependent increase in rCBF during aTDCS, even after accounting for individual differences in TDCS-induced electrical field strength. In contrast, finger movements led to robust activation of left M1-HAND before and after aTDCS. Conclusion: Short-duration bipolar aTDCS does not produce instantaneous dose-dependent rCBF increases in the targeted M1-HAND at conventional intensity ranges. Therefore, the regional hemodynamic response profile to short-duration aTDCS may not be suited to inform individual dosing of TDCS intensity.

In-Vitro Antidiabetic Effect of Ziziphus mucronata Leave Extracts

Background: Diabetes is a metabolic disorder characterized by hyperglycemia due to the body’s inability to produce insulin or inaction of the produced insulin or a combination of both. One antidiabetic therapeutic approach is to reduce gastrointestinal glucose production and absorption through the inhibition of carbohydrate digesting enzymes such as alpha-amylase as well as through the inhibition of hemoglobin glycosylation. Objective: This study sets out to evaluate the in vitro antidiabetic activity of Ziziphus mucronata extracts for their effect on alpha-amylase and glycosylation of hemoglobin. Methods: Successive gradient maceration of Z. mucronata leaves were carried out using Hexane, Acetone, Methanol and separately with water to obtain four (4) extracts labelled HE, AE, ME, and WE respectively. These were subjected to in vitro studies for their inhibitory effect on alpha-amylase and hemoglobin glycosylation, Standard laboratory methods were used to screen for phytochemicals of the most potent extract. Results: The result showed that AE, ME and WE extract exhibited a dose-dependent increase in percentage inhibition of both alpha-amylase and hemoglobin glycosylation. However, on a stricking note, the AE showed a more potent data result with percentage (%) potency of 71.02 at 1mg/ml, the lowest glucose concentration (of 25mg/ml) at 0.242nm as well as the highest hemoglobin glycosylation inhibitory mean concentration of 3.663nm after 72 hours. The AE of Z. mucronata (the most potent) revealed the presence of alkaloids, anthraquinone, glycosides, flavonoids, phenols, saponin, tanins and terpenoids. Conclusion: Thus, the Acetone extract is more likely to give a lead antidiabetic drug molecule of drug when further explored; which somewhat justify the folkloric claims of Z. mucronata leave as an antidiabetic. Keywords: Heamoglabin glycosylation, Alpha-amylase enzymes, Inhibition, Glucose.

Cytotoxic Synergism between a Proprietary Commiphora mukul Gum Extract GU-MCT810, 2-deoxy-D-glucose, and Metformin in Human Alveolar Rhabdomyosarcoma and Hepatoma Cell Lines In vitro

In this investigation we have analyzed the synergism for the cytotoxic effect of a proprietary guggul gum extract (GU), 2-deoxy-D-glucose (2-DG) and metformin (Met) in SJRH30 human alveolar rhabdomyosarcoma and HepG2 hepatoma cell lines in vitro. 2-DG and Met as single agents have weak cytotoxic effects in both cell lines. However, the combination of GU+2-DG, GU+Met and 2-DG+Met showed synergism for cytotoxic effect by CompuSyn analysis. Therefore, GU can be included in the combination of drugs involving 2-DG and Met to have synergistic effect. GU also showed a dose-dependent increase in cellular glucose uptake in HepG2 cells like the antidiabetic drug 2,4-thiozolidine dione (TZ). The demonstration of synergism of anticancer effects between GU, metformin and 2-DG, suggest that their mechanisms are in general complementary, though further studies are required to delineate the mechanism of GU, 2-DG and metformin combinations.

Gradual, but Not Sudden, Dose-Dependent Increase of ONJ Risk With Bisphosphonate Exposure: A Nationwide Cohort Study in Women With Osteoporosis

BackgroundA causal relationship of bisphosphonate (BP) exposure with osteonecrosis of the jaw (ONJ) has been reported; however, a definite dose-dependent risk remains to be elucidated beyond current vague recommendations of 4-year oral BP for ONJ risk increase.ObjectiveTo identify the effect of bisphosphonate cumulative dose on ONJ development in women with osteoporosis.MethodsA retrospective cohort study was designed using the National Health Insurance Service—National Health Screening database of Korea. Females over the age of 50 were diagnosed with osteoporosis based on the International Classification of Diseases 10th revision (ICD-10) codes (M80, M81, and M82) with bisphosphonate prescriptions. The cumulative dose of bisphosphonate was calculated using defined daily doses (DDD) to provide an accurate BP cumulative effect on ONJ occurrence. Osteonecrosis of the jaw was identified using both ICD-10 codes and related procedure codes. The incidence rates of ONJ and hazard ratios were estimated according to the bisphosphonate cumulative dose.ResultsAmong 74,491 included subjects, 190 cases of ONJ were identified. The incidence rate substantially increased after BP cumulative dose over 1 year (25.75 for DDD < 365, which increased to 53.43 for 365 ≤ DDD < 730). Compared to subjects with a cumulative dose of DDD < 365, subjects with a cumulative dose of 365 ≤ DDD < 730 had 2.36-fold hazard for developing ONJ (p < 0.001).ConclusionA bisphosphonate cumulative dose of more than 1 year had an increased risk of ONJ development. A gradual, but not sudden, dose-dependent increase in ONJ risk with BP exposure needs to be considered in providing the optimal BP treatment duration.

Linear Dose Response of Acrocentric Chromosome Associations (Aca) To Gamma Irradiation In Human Lymphocytes

Purpose: The frequency of acrocentric chromosome associations (ACA) was studied to determine the possible dose-response relation with low doses of gamma irradiation in lymphocytes. Methods: Peripheral blood collected from three healthy donors were irradiated with 0, 0.1, 0.25, 0.5, 0.75, and 1 Gy gamma radiation. Chromosomal preparations were made after 48 hrs culture as per the standard guidelines. Results: The average number of ACA and ACA % were increased significantly with an increase in a dose. The D-G and D-D type of association was most prominent and showed a dose-dependent increase. The ACA frequency in irradiated lymphocytes showed an increase concerning the dose. The fitted regression equation was y=0.4759x+0.1663 (R2=0.9635; p=0.0005). An assessment of dicentric chromosomes (DC) was carried for the same slides. The correlation curve was prepared for ACA frequencies versus DC frequencies, resulting in a regression equation as y=8.659x+0.2.37 (R2=0.8275; p=0.0119). Conclusion: Our results showed an increase in frequencies of ACA in irradiated lymphocytes with an increase in radiation dose and followed a similar linear trend with DC frequency, thus, ACA may serve as a candidate cytogenetic biomarker for radiation biodosimetry especially for low radiation doses.

Mycobacterium tuberculosis senses host Interferon- gamma via the membrane protein MmpL10

Mycobacterium tuberculosis (Mtb) is one of the most successful human pathogens and remains a leading cause of death from infectious disease. Interferonγ (IFNγ) is a central regulator of the immune defense against Mtb. Several cytokines have been shown to increase virulence of other bacterial pathogens, leading us to investigate whether IFNγ has a direct effect on Mtb. We found that both recombinant and T cell derived IFNγ rapidly induced a dose dependent increase in the oxygen consumption rate (OCR) of Mtb, consistent with increased bacterial respiration. This was also observed in clinical strains, but not in the vaccine strain Bacillus Calmette Guerin (BCG), and did not occur for other cytokines tested, including TNFα. IFNγ binds to the cell surface of intact Mtb, but not BCG, whilst TNFα binds to neither. Mass spectrometry analysis identified mycobacterial membrane protein large 10 (MmpL10) as the transmembrane binding partner. Consistent with this, IFNγ binding and the OCR response was absent in a Mtb Δmmpl10 strain and restored by complementation of the mutant strain. RNA-sequencing of IFNγ exposed Mtb revealed a distinct transcriptional profile, including genes involved in virulence and cholesterol catabolism. Finally, exposure of Mtb cells to IFNγ resulted in sterilization of bacilli treated with isoniazid (INH), indicating clearance of phenotypically resistant bacteria that persist in the presence of INH alone. Our data suggest a novel mechanism allowing Mtb to respond to host immune activation that may be important in the immunopathogenesis of TB and have use in novel eradication strategies.

703 Favorable pre-clinical safety profile of the novel not-alpha IL-2 agonist ANV419 supports first in human clinical development

BackgroundANV419 is a novel interleukin-2 (IL-2)/anti-IL-2 fusion protein with preferential signaling through the IL-2 beta/gamma receptor that induces selective proliferation of CD8 T cells and NK cells in vivo for the treatment of cancer. The safety and pharmacodynamic effects of ANV419 were studied in a 4-week cynomolgus monkey GLP study to support the ongoing PhI dose escalation clinical trial.MethodsANV419 was administered by i.v. injection over 1 min at doses of 0.03, 0.1, 0.3 mg/kg, or vehicle control on days 1 and 15 of the 29-day study. Assessments included body weight, blood pressure, hematology, clinical pathology, serum cytokines, immunophenotyping, histopathology, and pharmacokinetics.ResultsThe pharmacokinetics of ANV419 were characterized by target mediated disposition, with a half-life of approximately 24h at concentrations not affected by target mediated clearance. Dose-dependent increases in WBC were observed after each injection, driven by preferential expansion of CD8 T cells and NK cells over Tregs. NK cells were more sensitive to ANV419 than CD8 T cells reaching maximal proliferation in blood at 0.03 mg/kg vs. 0.3 mg/kg for CD8 T cells. Hematological changes included: transient dose-dependent increase in basophils; elevation in eosinophils, up to 2.2-fold above control animals at > 0.03 mg/kg, remaining within the normal range for cynomolgus monkeys (<1.94 G/L); minor decrease in platelets at day 4 after each injection. There were no relevant treatment-related changes in inflammatory serum cytokines (IL-1b, IL-5, IL-6, IL-8, IFNg, TNFa, GM-CSF). A mild systemic inflammatory response was observed at 0.3 mg/kg evidenced by a transient increase of CRP on days 4 and 19, preceded after the first injection by a slight dose dependent increase in IL-1RA at 4h post injection, and an increase in IL-10 at 24h post treatment at 0.3mg/kg. No significant changes in body weights or blood pressure and no signs of capillary leak were observed during the entire study.A multi-part PhI dose-escalation study of ANV419 has been initiated in cancer patients. In the part A single patient escalation cohort, two patients have been dosed Q2W multiple times with 0.003mg/kg and 0.006mg/kg respectively with the expected PD profile and no DLT observed.ConclusionsConsistent findings, relating to expected effects of ANV419 as a not-alpha IL-2 agonist, demonstrated a favorable tolerability and safety profile at pharmacodynamically relevant doses that strongly support its translational development in cancer patients to identify clinical benefits.

IL-6 induced enhanced clearance of proANP and ANP by insulin-degrading enzyme in T1DM mouse

Cardiovascular disease (CVD) is the prevalent cause of morbidity and mortality in type I diabetes mellitus (T1DM) worldwide. However, the pathophysiological mechanisms underlying the relationship between CVD, CVD risk factors, and T1DM have not yet been sufficiently explored. Here we reported that insulin-degrading enzyme (IDE) effectively degrades the precursor of atrial natriuretic peptide (proANP) intracellular in HEK293T cells. Pro-inflammatory cytokine IL-6 elicited a significant dose-dependent increase in IDE protein expression. Inhibition of ERK/MAPK signaling pathway with selumetinib abolished IL-6-stimulated increase in IDE protein level and deceased in ANP secretion in H9C2 cells. Importantly, the T1DM mouse model displayed lower proANP in the heart and ANP in serum, due to increased IDE expression and activity. Our outcomes suggest a novel role of IL-6 on ANP metabolism via IDE and provide the possibilities for new potential therapeutic strategies for diabetes-related cardiovascular complications.

Induced antigen-binding polyspecificity in human serum IgA

Recent studies have shown that polyspecific antibodies play an important role in the frontline defense against the dissemination of pathogens in the pre-immune host. Interestingly, antigen-binding polyspecificity can not only be inherent, but also acquired post-translationally. The ability of individual monoclonal IgE and IgG antibodies to acquire polyspecificity following contact with protein-modifying agents has been studied in detail. However, to the best of our knowledge this property of human IgA has previously been described only cursorily. In the present study pooled human serum IgA and two monoclonal IgA antibodies were exposed to buffers with acidic pH, to free heme or to pro-oxidative ferrous ions, and the antigen-binding behavior of the native and modified IgA to viral and bacterial antigens were compared using Western blotting and ELISA. We observed a dose-dependent increase in reactivity to several bacterial extracts and to pure viral antigens. This newly described property of IgA may have therapeutic potential as has already been shown for pooled IgG with induced polyspecificity.

Evaluation Of The Mutagenic Potential Of The Polyphenol Extract From Blueberries In The Ames Test

In this research, mutagenic properties of blueberry polyphenol extract were studied in gene mutation induction test (Ames test) on four strains of Salmonella typhimurium TA98, TA100, TA1535, TA1537. None of the strains of Salmonella typhimurium showed statistically reliable dose-dependent increase in number of revertant colonies in the presence of investigated drug in the studied dose range from 4,0 to 40,0 mg/ml relative to baseline of spontaneous mutations. The blueberry extract does not have any mutagenic activity in the researched dose range in relation to TA98, TA100, TA1535, TA1537 strains of Salmonella typhimurium.