BACKGROUND

To examine chronic viral hepatitis (CVH) as a risk factor for hepatotoxicity during isoniazid (INH) treatment for latent tuberculosis infection (LTBI).

METHODS

A search of MEDLINE (1966-May 2008) was conducted using the terms 'tuberculosis', 'antitubercular', 'therapeutics', 'treatment', 'prevention', 'prophylaxis', 'hepatitis', 'toxic hepatitis', 'hepatotoxic', 'liver' and 'injury'. Peer-reviewed, English-language articles describing the relationship between a history of CVH and occurrence of hepatotoxicity during LTBI treatment were selected. We limited CVH diagnoses to reports with positive serological test or biopsy for hepatitis B or C. Risk ratios and 95% confidence intervals were abstracted or derived.

RESULTS

We reviewed 486 abstracts, and 11 studies met the selection criteria. Populations included in the studies were the general population (n = 6) and transplant recipients (n = 5). The variability in study designs and case finding practices precluded performing a quantitative meta-analysis. Two studies of former or current drug users reported a consistent, positive association between chronic hepatitis C infection and INH hepatotoxicity. Other risk ratios did not significantly or consistently show any association between CVH in patients treated for LTBI and the development of INH hepatotoxicity.

CONCLUSIONS

Owing to the limited number of published papers, CVH was not established as a risk factor for INH hepatotoxicity during LTBI treatment. Controlled studies are needed to define the safety and tolerability of LTBI treatment in those with CVH and to provide an evidence base for recommendations for LTBI treatment in persons with CVH.

Isoniazid (INH) was studied with regard to its electrochemical treatment on a strongly alkaline solution of a poly (3,4-ethylenedioxythiophene)-modified gold electrode, using both cyclic voltammetric and controlled potential techniques. Electrocatalytic performance measurements of this composite electrode toward oxidation of INH exhibited an increase of 4-folds in oxidation peak densities compared to the bare gold electrode. Central composite design method was used to obtain optimum experimental conditions, and used critical parameters (pH (A), scan rate (mV/s, B) and temperature (C, C) to assess the effects on amperometric response. Optimum experimental conditions were achieved by using a pH of 9.2 with a scan rate of 260 mV/s and a temperature of 30 C. Under these circumstances, a good linear relationship was observed between peak current densities and INH concentration in the range of 0.05-2 μM, with correlation coefficient of 0.9998. Furthermore, the method was very sensitive (limit of quantitation, 0.0043 μM), accurate (relative error, -5.65 to 4.03) and precise (relative standard deviation %, ≤ 7.97). The method was also applied to determine INH in pharmaceutical formulations, and statistically compared the results with the official method using the two one-sided equivalence test; the results were in good agreement with those obtained by the official and reported methods.

BACKGROUND

Vaginal lubrication, an indicator of sexual arousal and tissue health, increases significantly during genital sexual arousal. Adrenergic alpha-receptors (AR) are an important regulator of genital physiological responses involved in mediating vascular and nonvascular smooth muscle contractility; the role of β-AR in sexual arousal, however, has not yet been investigated.

OBJECTIVE

The goal of this study was to reveal the functional role of β-AR in modulating vaginal lubrication during sexual arousal and the mechanisms underlying the process.

METHODS

The effects of adrenaline on vaginal epithelial ion transport, intracellular cyclic adenosine monophosphate (cAMP) content ([cAMP]i ), and vaginal lubrication were investigated using short-circuit current (ISC ) of rat vaginas incubated in vitro, enzyme-linked immunosorbent assay (ELISA), and measurement of vaginal lubrication in vivo, respectively. The expressions of β-AR in vaginal epithelium were analyzed by reverse transcription-polymerase chain reaction, western blot, and immunofluorescence.

METHODS

Changes of ISC responses; mRNA, protein expressions and localization of β-AR; [cAMP]i ; vaginal lubrication.

RESULTS

Serosal application of adrenaline induced an increase of ISC across rat vaginal epithelium that blocked by propranolol, a <em>β</em>-AR antagonist, rather than phentolamine, an α-AR antagonist. <em>β</em>1/2-AR were both present in rat and human vaginal epithelial cells. Removing Cl(-) or application of CFTR(inh) -172, an inhibitor of cystic fibrosis transmembrane conductance regulator (CFTR), abolished adrenaline-induced ISC responses. The elevated levels of [cAMP]i induced by adrenaline were prevented by the pretreatment with propranolol. Vaginal lubrication measured in vivo showed that adrenaline or pelvic nerve stimulation caused a marked increase in vaginal lubrication, whereas pretreatment with propranolol or CFTR(inh) -172 reduced the effect.

CONCLUSIONS

Activation of epithelial β-AR facilitates vaginal lubrication during sexual arousal by stimulating vaginal epithelial Cl(-) secretion in a cAMP-dependent pathway. Thus, vaginal epithelial β-AR might be another regulator of vaginal sexual arousal responses.

<p><div>(<em>b</em>)BACKGROUND & AIMS</<em>b</em>)</div>SLC26A3 (DRA) is an electroneutral Cl^-/HCO<su<em>b</em>)3</su<em>b</em>)^- exchanger that is present in the apical domain of multiple intestinal segments. An area that has continued to <em>b</em>e poorly understood is related to DRA regulation in acute adenosine 3',5'-cyclic monophosphate (cAMP)-related diarrheas, in which DRA appears to <em>b</em>e <em>b</em>oth <em>inh</em>i<em>b</em>ited as part of NaCl a<em>b</em>sorption and stimulated to contri<em>b</em>ute to increased HCO<su<em>b</em>)3</su<em>b</em>)^- secretion. Different cell models expressing DRA have shown that cAMP <em>inh</em>i<em>b</em>its, stimulates, or does not affect its activity.</p><p><div>(<em>b</em>)METHODS</<em>b</em>)</div>This study re-evaluated cAMP regulation of DRA using new tools, including a successful knockout cell model, a specific DRA <em>inh</em>i<em>b</em>itor (DRA<su<em>b</em>)<em>inh</em></su<em>b</em>)-A250), specific anti<em>b</em>odies, and a transport assay that did not rely on nonspecific <em>inh</em>i<em>b</em>itors. The studies compared DRA regulation in colonoids made from normal human colon with regulation in the colon cancer cell line, Caco-2.</p><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>DRA is an apical protein in human proximal colon, differentiated colonoid monolayers, and Caco-2 cells. It is glycosylated and appears as 2 <em>b</em>ands. cAMP (forskolin) acutely stimulated DRA activity in human colonoids and Caco-2 cells. In these cells, DRA is the predominant apical Cl^-/HCO<su<em>b</em>)3</su<em>b</em>)^- exchanger and is <em>inh</em>i<em>b</em>ited <em>b</em>y DRA<su<em>b</em>)<em>inh</em></su<em>b</em>)-A250 with a median <em>inh</em>i<em>b</em>itory concentration of 0.5 and 0.2 μmol/L, respectively. However, there was no effect of cAMP in HEK293/DRA cells that lacked a cystic fi<em>b</em>rosis transmem<em>b</em>rane conductance regulator (CFTR). When CFTR was expressed in HEK293/DRA cells, cAMP also stimulated DRA activity. In all cases, cAMP stimulation of DRA was not <em>inh</em>i<em>b</em>ited <em>b</em>y CFTR<su<em>b</em>)<em>inh</em></su<em>b</em>)-172.</p><A<em>b</em>stractText>DRA is acutely stimulated <em>b</em>y cAMP <em>b</em>y a process that is CFTR-dependent, <em>b</em>ut appears to <em>b</em>e one of multiple regulatory effects of CFTR that does not require CFTR activity.</A<em>b</em>stractText>

Isoniazid (INH) and Rifampicin (RIF) are known hepatotoxic agents. We compared the efficacy of Spirulina fusiformis and its active components vitamin B12 and beta-carotene in attenuating INH and RIF induced hepatotoxicity. We also tried to elucidate the inflammatory mechanism behind anti-tuberculosis drug induced hepatotoxicity. INH and RIF were administered to Wistar albino rats for 28 days to induce hepatotoxicity. S. fusiformis, vitamin B12, and beta-carotene were co-administered with INH and RIF and their hepatoprotective, antioxidant, and immunomodulatory roles were studied through blood and liver analysis. Changes induced by INH and RIF in antioxidants, cytokines (IL-6 and IL-10) and expression of Nuclear Factor-κB (NF-κB) and Nitric Oxide Synthase (iNOS) were also studied. Supplement treatment caused restoration of liver function parameters to normal levels along with reversal of inflammatory changes in IL-6 and IL-10 levels. Liver PCNA, iNOS, and NF-κB expression were reduced in the supplement treated tissues compared to INH and RIF treated rats as evidenced by immunohistochemistry and quantitative PCR. Correlation of IL-6 levels, PCNA, and iNOS with NF-κB showed its pivotal role in the inflammatory process. Study shows the pivotal role of NF-kB and the equivalence in antioxidant efficacy of vitamin B12 and beta-carotene compared to Spirulina fusiformis. J. Cell. Biochem. 118: 3825-3833, 2017. © 2017 Wiley Periodicals, Inc.

Misfolding of the protein α-synuclein (αSyn) into amyloid fibrils plays a central role in the development of Parkinson's disease. Most approaches for the inhibition of αSyn fibril formation are based on stabilizing the native monomeric form of the protein or destabilizing the fibrillized misfolded form. They require high concentrations of inhibitor and therefore cannot be easily used for therapies. In this work, we designed an inhibitor (Inh-β) that selectively binds the growing ends of αSyn fibrils and creates steric hindrance for the binding of monomeric αSyn. This approach permits the inhibition of fibril formation at Inh-β concentrations (IC50 =850 nm) much lower than the concentration of monomeric αSyn. We studied its kinetic mechanism in vitro and identified the reactions that limit inhibition efficiency. It is shown that blocking of αSyn fibril ends is an effective approach to inhibiting fibril growth and provides insights for the development of effective inhibitors of αSyn aggregation.

The continuous discharge of antibiotic pharmaceuticals from incomplete wastewater treatment processes into receiving water bodies has become a matter of both scientific and public concern as antibiotics may exert adverse influences on non-target organisms. In this study, the occurrence of seven most commonly prescribed antibiotics belonging to four therapeutic classes of β-lactams, cephalosporins, macrolides, and fluoroquinolones were investigated in the effluent of two wastewater treatment plants (WWTPs) and two river waters: Firozabad Ditch (receiving effluent) and Kan River (not receiving effluent) in Tehran, Iran. In 2016, average consumption rate of target antibiotics in Tehran province was evaluated based on Anatomical Therapeutic chemical (ATC)/Defined Daily Dose (DDD) system and reported as DDD/1000 inh/day. The highest consumption rate was for amoxicillin (128017.6 mg/1000 inhabitants/day), whereas it remained lower for other compounds (amoxicillin > cefixime > azithromycin > ciprofloxacin > cephalexin > erythromycin > penicillin). Ciprofloxacin (79.62 mg/1000 inh/d) and cephalexin (209.51 mg/inh/d) with highest mass loads were evaluated in the influent of WWTP A and WWTP B, respectively. Ciprofloxacin (24.87 mg/1000 inh/d) and cefixime (90.45 mg/1000 inh/d) were the highest evaluated mass loads in the effluent of Ekbatan wastewater treatment plant (WWTP A) and Tehran Southern wastewater treatment plant (WWTP B), respectively. The calculated risk quotients showed that six out of seven target antibiotics posed a high risk to algae (M. aeruginosa and P. subcapitata) and bacteria (P. putida) in the effluent of WWTPs and the rivers wherein amoxicillin and penicillin posed a higher risk than other antibiotics occurring due to their lowest PNEC.

OBJECTIVE

Ovarian inhibins (INH) are hormones participating in the regulation of gametogenesis. We monitored inhibin A and B levels in serum (S) and follicular fluids (FF), depending on the type of fertility failure and treatment outcome.

METHODS

We examined INHA and B levels in S and FF of 72 women undergoing ovarian stimulation for in vitro fertilization, including embryo transfer We took serum samples at the time of egg collection (S1), embryo transfer (S2), and diagnostics of early pregnancy (S3). FF samples were obtained during egg collection. INH A and B levels were measured by ELISA set kit in all media.

RESULTS

Healthy women had median of INHA S1 592.02pg/ml INHA S2 593.58pg/ml, INHA S3 15.17pg/ml and INH B S1 242.46pg/ml, INH B S2 and INH B S3 zero levels. Women with ovarian disorders had significantly lower levels of INH A S1 and INH A S2 (p<0.05). Women with polycystic ovaries had significant higher INH B S2 levels (p<0. 05). No statistically significant differences were found in women with endometriosis. Presence of oocyte in the dominant follicle positively correlated with INH B FF levels (p<0.05).

CONCLUSIONS

WE confirmed differences in the levels of inhibins in sera depending on type of fertility failure. Inhibin B better reflected the presence of an oocyte. The potential paracrine role of inhibins needs to be examined to improve preparation for the in vitro fertilization treatment (IVF).

Building on studies showing that ischemia-reperfusion-(I/R)-injury is complement dependent, we tested links among complement activation, transplantation-associated I/R injury, and murine cardiac allograft rejection. We transplanted BALB/c hearts subjected to 8-h cold ischemic storage (CIS) into cytotoxic T-lymphocyte associated protein 4 (CTLA4)Ig-treated wild-type (WT) or c3-/- BB (cfb-/- ) recipients (n = 4-6/group, p < 0.05, mbl1-/- mbl2-/- vs. cfb-/- ), implicating the MBL (not alternative) pathway. To pharmacologically target MBL-initiated complement activation, we transplanted BALB/c hearts subjected to 8-h CIS into CTLA4Ig-treated WT BINH). Remarkably, peritransplantation administration of C1-INH prolonged graft survival (MST >60 days, p < 0.05 vs. controls, n = 6) and prevented CI-induced increases in donor-reactive, IFNγ-producing spleen cells (p < 0.05). These new findings link donor I/R injury to T cell-mediated rejection through MBL-initiated, complement activation and support testing C1-INH administration to prevent CTLA4Ig-resistant rejection of deceased donor allografts in human transplant patients.

An enzyme-free fluorometric assay is described for the determination of zearalenone (ZEN). The method combines (a) catalyzed hairpin assembly (CHA), (b) ultrahigh fluorescent light-up G-rich DNA sequences in proximity to silver nanoclusters (Ag NCs), and (c) the use of aptamers (Apt). In the presence of ZEN, the inhibit sequence (Inh) is released from the aptamer-trigger sequence (Apt-T) via the binding of ZEN and the aptamer of Apt-T. The free Apt-T acts as a switch that opens the hairpins H1 and H2 to generate H1-H2 complex. The released Apt-T is available to trigger the next round of CHA between H1 and H2. Finally, the hybridization between H1 and the Ag NCs probe (P) causes the G-rich sequence to be in close proximity to the dark Ag NCs encapsulated by P. This leads to highly efficient lighting up of the Ag NCs and the production of amplified fluorescence with excitation/emission peaks at 575/628 nm. Under the optimized conditions, a linear correlation was observed with concentrations ranging from 1.3 pg mL^-1 to 100 ng mL^-1, and the limit of detection was 0.32 pg mL^-1 (at S/N = 3). The method was successfully validated by analyzing maize and beer for levels of ZEN after a simple sample preparation procedure. Graphical abstractSchematic of the assay. The inhibit sequence (Inh) is released from aptamer-trigger sequence (Apt-T) via binding of ZEN and aptamer. The free Apt-T triggers catalyzed hairpin assembly (CHA).G-rich DNA is in proximity to silver nanoclusters (Ag NCs) and fluorescence intensity increases to detect ZEN.

OBJECTIVE

To explore the value of serum inhibin measurement in predicting ovarian response in infertile patients undergoing controlled ovarian hyperstimulation (COH) for in vitro fertilization (IVF) cycle.

METHODS

A total of 37 patients undergoing their first cycles of IVF treatment were studied and serum estradiol (E(2)), progesterone (P), inhibin (INH) B, INHA levels were detected on days 2 - 4 and 21 of the previous cycle, on day 5 of gonadotropin therapy (day 5), hCG injection day, oocyte retrieval day and at middle luteal phase of the IVF cycle. According to number of oocytes, peak E(2) concentration and presence or absence of ovarian hyperstimulation syndrome (OHSS), the 37 patients were allocated into two groups: poor ovarian response group (group A), normal and high response group (group B). Characteristics of the treatment cycle and INH, E(2), P levels were analyzed. A multiple logistic regression model and receiver operating characteristic (ROC) analysis were then used for all possible prognostic variables to evaluate the value of different hormone in predicting ovarian response. The sensitivity and specificity were calculated at the discriminating cut-off point.

RESULTS

(1) In group A, basal serum INHB level, INHB and INHA levels on day 5 of recombinant FSH (rFSH) stimulation were significantly lower than group B (P < 0.05), and basal serum INHB levels were (37 +/- 35), (91 +/- 90) ng/L respectively, INHB levels on day 5 of rFSH stimulation were (194 +/- 157), (2254 +/- 4765) ng/L respectively, INHA levels on day 5 of rFSH stimulation were (36 +/- 35), (91 +/- 90) ng/L respectively in groups A and B. (2) There was positive correlation between oocytes number and basal inhibin B and dynamic serum inhibin B levels, inhibin A and E(2) levels of rFSH therapy (INHB: r = 0.39 - 0.67, P < 0.05; INHA: r = 0.43 - 0.59, P < 0.01; E(2): r = 0.50 - 0.60, P < 0.01). (3) ROC analysis: day 5 inhibin B had the largest area under curve (AUC) 0.948 (95% CI: 0.871 - 1.025), indicating that a threshold of 400 ng/L gave 82.8% sensitivity, and 99.1% specificity.

CONCLUSIONS

The basal INHB levels and INHB measured on day 5 of rFSH stimulation may offer accurate and early prediction of ovarian response. Day 5 INHB levels are the best predictor of ovarian response.

<p><div>(<em>b</em>)Background</<em>b</em>)</div>Hereditary angioedema (HAE) is a de<em>b</em>ilitating disorder resulting from C1-esterase inhi<em>b</em>itor (C1-<em>INH</em>) deficiency. In the COMPACT phase 3 study the prophylactic use of a su<em>b</em>cutaneous C1 inhi<em>b</em>itor (C1-<em>INH</em> [SC], HAEGARDA^®, CSL Behring) twice weekly significantly reduced the frequency of acute edema attacks. Analysis of treatment effects <em>b</em>y su<em>b</em>groups, onset of effect, and other exploratory analysis have not <em>b</em>een reported.</p><A<em>b</em>stractText>This is a post hoc exploratory analysis on data from the randomized, place<em>b</em>o-controlled COMPACT study. 90 patients with C1-<em>INH</em>-HAE were randomized to 1 of 4 treatment sequences: C1-<em>INH</em> (SC) 40 or 60 IU/kg of <em>b</em>ody weight twice weekly for 16 weeks, preceded or followed <em>b</em>y a place<em>b</em>o period. The pre-specified primary efficacy endpoint was the time-normalized num<em>b</em>er of HAE attacks, and pre-specified secondary efficacy endpoints were the percentage of patients with a certain treatment response (≥ 50% reduction on C1-<em>INH</em> (SC) versus place<em>b</em>o in the time-normalized num<em>b</em>er of attacks) and the time-normalized num<em>b</em>er of use of rescue medication. Pre-specified exploratory endpoints included severity of attacks, alone and com<em>b</em>ined with rescue medication use. Post hoc analyses included exploration of onset of effect and clinical assessment of patients with < 50% of response.</A<em>b</em>stractText><A<em>b</em>stractText>Su<em>b</em>group findings <em>b</em>y various patient characteristics showed a consistent preventive effect of C1-<em>INH</em> (SC). In a post hoc analysis of attacks, the onset of the preventive effect within the first 2 weeks after treatment initiation in COMPACT showed that 10/43 patients (23%) experienced attacks of any severity with 60 IU/kg versus 34/42 patients (81%) with place<em>b</em>o. The need for rescue medication was tenfold lower with 60 IU/kg (35 treated attacks) versus place<em>b</em>o (358 treated attacks). A qualitative analysis of the 4 patients treated with 60 IU/kg and with < 50% reduction of attacks demonstrated a reduction in severity of attacks, rescue medication use, and symptom days which was considered a clinically meaningful treatment effect.</A<em>b</em>stractText><p><div>(<em>b</em>)Conclusions</<em>b</em>)</div>C1-<em>INH</em> (SC) prophylaxis demonstrated a preventive treatment effect with evidence of <em>b</em>enefit within 2 weeks. A consistent treatment effect at recommended C1-<em>INH</em> (SC) dosing was evident in all su<em>b</em>groups of patients with type I/II HAE and <em>b</em>y various measures of disease and treatment <em>b</em>urden.<i>Trial registration</i> EU Clinical Trials Register, 2013-000916-10, Registered 10 Decem<em>b</em>er 2013, https://www.clinicaltrialsregister.eu/ctr-search/trial/2013-000916-10; ClinicalTrials.gov Register, NCT01912456, Registered 31 July 2013, https://clinicaltrials.gov/ct2/show/NCT01912456.</p>

BACKGROUND

Schiff bases have a broad spectrum of biological activities like antiinflammatory, analgesic, antimicrobial, anticonvulsant, antitubercular, anticancer, antioxidant, anthelmintic and so forth. Thus, after a thorough perusal of literature, it was decided to conjugate benzothiazol-2-ylamine/thiazolo [5, 4-b] pyridin-2-ylamine with aromatic and heteroaromatic aldehydes to get a series of Schiff bases.

OBJECTIVE

Synthesis, characterization, in-silico toxicity profiling and anticonvulsant activity of the Schiff bases of Benzothiazol-2-ylamine and Thiazolo [5, 4-b] pyridin-2-ylamine.

METHODS

Aniline/4-aminopyridine was converted to the corresponding thiourea derivatives, which were cyclized to obtain benzothiazol-2-ylamine/thiazolo [5, 4-b] pyridin-2-ylamine. Finally, these were condensed with various aromatic and heteroaromatic aldehydes to obtain Schiff bases of benzothiazol-2-ylamine and thiazolo [5, 4-b] pyridin-2-ylamine. The synthesized compounds were characterized and screened for their anticonvulsant activity using maximal electroshock (MES) test and isoniazid (INH) induced convulsions test. In-silico toxicity profiling of all the synthesized compounds was done through "Lazar" and "Osiris" properties explorer.

RESULTS

Majority of the compounds were more potent against MES induced convulsions than INH induced convulsions. Schiff bases of benzothiazol-2-ylamine were more effective than thiazolo [5, 4-b] pyridin-2-ylamine against MES induced convulsions. The compound benzothiazol-2-yl-(1H-indol-2-ylmethylene)-amine (VI) was the most potent member of the series against both types of convulsions.

CONCLUSIONS

Compound VI exhibited the most significant activity profile in both the models. The compounds did not exhibit any carcinogenicity or acute toxicity in the in-silico studies. Thus, it may be concluded that the Schiff bases of benzothiazol-2-ylamine exhibit the potential to be promising and non-toxic anticonvulsant agents.

BACKGROUND

Common deterrents to insulin therapy for both physicians and patients are the complexity and burden of daily injections. In January 2006, the first inhaled human insulin (INH, Exubera (insulinhuman [rDNA origin])InhalationPowder) was approved for use in adult patients with type 1 diabetes mellitus (T1DM) or type 2 diabetes mellitus (T2DM) in the United States and European Union. Results from the INH clinical trial program have shown comparable efficacy of INH to subcutaneous (s.c.) insulin and superior efficacy versus oral antidiabetic agents; thus providing effective glycemic control in adult patients with T2DM without the requirement for preprandial injections. However, because subjects in those trials were randomized to either INH or an alternative, the studies could not estimate the effect of INH on patient acceptance of insulin therapy. Therefore, traditional study designs cannot provide answers to important and practical questions regarding real world effectiveness, which is influenced by psychological and other access barriers.

METHODS

To overcome these limitations, the Real World Trial was designed to estimate the effect of the availability of INH as a treatment option for glycemic control. A total of approximately 700 patients from Canada, France, Germany, Italy, Spain, United Kingdom, and the United States with T2DM poorly controlled by oral agent therapy will be randomized to two different treatment settings. Patients and clinicians in both groups (A & B) may choose from all licensed therapies for diabetes including SC insulin delivered by pens; INH will be an additional treatment option only available in Group A. The Real World Trial (Protocol A2171018) has been registered with ClincalTrials.gov, registration id NCT00134147.

RESULTS

The primary outcome for the trial will be the difference in mean glycosylated hemoglobin (HbA1c) at 6 months between groups. The design was based on a preceding feasibility study examining the theoretical effects of inhaled insulin availability on treatment choice in 779 patients. In that study, patients were three times more likely to choose insulin therapy when inhaled insulin was available.

CONCLUSIONS

Innovations in study designs may provide an opportunity to reveal unbiased answers to important treatment questions that are more relevant to prescribers, funding agencies, and healthcare policymakers.

Sixty-two multidrug resistant Salmonella enterica serovar Typhimurium strains isolated from 255 clinical strains collected in Southern Italy in 2006-2008 were characterised for antimicrobial resistance genes, pulsotype, and phage type. Most strains (83.9%) were resistant to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline (ACSSuT) encoded in 88.5% by the Salmonella genomic island (SGI1) and in 11.5% by the InH-like integron (bla OXA-30-aadA1) and catA1, sul1, and tet(B) genes. STYMXB.0061 (75%) and DT120 (84.6%) were the prevalent pulsotype and phage type identified in these strains, respectively. Five other resistance patterns were found either in single or in a low number of isolates. The pandemic clone DT104 (ACSSuT encoded by SGI1) has been identified in Italy since 1992, while strains DT120 (ACSSuT encoded by SGI1) have never been previously reported in Italy. In Europe, clinical strains DT120 have been reported from sporadic outbreaks linked to the consumption of pork products. However, none of these strains were STYMXB.0061 and SGI1 positive. The prevalent identification and persistence of DT120 isolates would suggest, in Southern Italy, a phage type shifting of the pandemic DT104 clone pulsotype STYMXB.0061. Additionally, these findings raise epidemiological concern about the potential diffusion of these emerging multidrug resistant (SGI linked) DT120 strains.

Objectives: Interferons (IFNs) play multifunctional roles in host defense against infectious diseases by inducing IFN-stimulated genes (ISGs). However, little is known about how ISGs regulate host immune response to Mycobacterium tuberculosis (Mtb) infection, the major cause of tuberculosis (TB).

Methods: We thus profiled the potential effects and mechanisms of eight Mtb-induced ISGs on Mtb infection by RNA interference in human macrophages (Mφs) derived from peripheral blood monocytes (hMDMs) and THP-1 cell line derived Mφs (THP-1-Mφs).

Results: MxA silencing significantly decreased intracellular Mtb infection in Mφs. Mechanistically, MxA silencing promoted inflammatory cytokines IL-1β, IL-6 and TNF-α production, and induced NF-κB p65 activation. Pharmacological inhibition of NF-κB p65 activation or gene silencing of NF-κB p65 blocked the increased production of IL-1β, IL-6 and TNF-α and restored Mtb infection by MxA silencing. Furthermore, pharmacological inhibition of TAK1 and IKKα/β blocked NF-κB p65 activation and subsequent production of pro-inflammatory cytokines by MxA silencing. Isoniazid (INH) treatment and MxA silencing could promote TAK1-IKKα/β-NF-κB signaling pathway activation and combat Mtb infection independently.

Conclusions: Our results reveal a novel role of MxA in regulating TAK1-IKKα/β-NF-κB signaling activation and production of antimicrobial inflammatory cytokines upon Mtb infection, providing a potential target for clinical treatment of TB.

Keywords: IκB kinase α/β (IKKα/β); Macrophages (Mφs); Mycobacterium tuberculosis (Mtb); Myxovirus resistance protein 1 (MxA); Nuclear factor-κB (NF-κB); TGF-β-activating kinase 1 (TAK1).

We aimed to describe incidence, trends of tuberculosis (TB) over 18 years and to evaluate the impact of the BCG vaccine after four decades of immunization program according to three protocols. We performed a cohort study including declared cases in Monastir from January 1, 2000 to December 31, 2017. We reported 997 cases of TB. The predominant site was pulmonarylocalization (n = 486). The age standardized incidence of pulmonary and lymph node TB per 100,000 inh were 5.71 and 2.57 respectively. Trends were negative for pulmonary TB (PTB) (b = - 0.82; r = -0.67; p<10-3) and positive for lymph node localization (b = 1.31; r = 0.63; p<10-3). We had not notified cases of HIV associated with TB. Crude incidence rate (CIR) of PTB per 100,000 inh was 8.17 in Non-Vaccinated Cohort (NVC) and 2.85 in Vaccinated Cohort (VC) (p < 0.0001). Relative risk reduction (RRR) of BCG vaccination was 65.1% (95%CI:57.5;71.4) for pulmonary localization and 65% (95%CI:55; 73) for other localizations. We have not established a significant RRR of BCG vaccination on lymph node TB. Protocol 3 (at birth) had the highest effectiveness with a RRR of 96.7% (95%CI: 86.6%; 99.2%) and 86% (95%CI:71%;91%) in patients with PTB and other localizations TB respectively. In Cox regression model the HR was 0.061 (95% CI 0.015-0.247) for PTB and 0.395 (95% CI 0.185-0.844) for other localizations TB in patients receiving protocol 3 compared to NVC. For lymph-node TB, HR was 1.390 (95% CI 1.043-1.851) for protocol 1 and 1.849 (95% CI 1.232-2.774) for protocol 2 compared to NVC. Depending on the three protocols, the BCG vaccine had a positive impact on PTB and other TB localizations that must be kept and improved. However, protocols 1 and 2 had a reverse effect on lymph node TB.

We report GSK3011724A (DG167) as a <em>b</em>inary inhi<em>b</em>itor of <em>β</em>-ketoacyl-ACP synthase (KasA) in <i>Myco<em>b</em>acterium tu<em>b</em>erculosis</i> Genetic and <em>b</em>iochemical studies esta<em>b</em>lished KasA as the primary target. The X-ray crystal structure of the KasA-DG167 complex refined to 2.0-Å resolution revealed two interacting DG167 molecules occupying nonidentical sites in the su<em>b</em>strate-<em>b</em>inding channel of KasA. The <em>b</em>inding affinities of KasA to DG167 and its analog, 5g, which <em>b</em>inds only once in the su<em>b</em>strate-<em>b</em>inding channel, were determined, along with the KasA-5g X-ray crystal structure. DG167 strongly augmented the <i>in vitro</i> activity of isoniazid (<em>INH</em>), leading to synergistic lethality, and also synergized in an acute mouse model of <i>M. tu<em>b</em>erculosis</i> infection. Synergistic lethality correlated with a unique transcriptional signature, including upregulation of oxidoreductases and downregulation of molecular chaperones. The lead structure-activity relationships (SAR), pharmacokinetic profile, and detailed interactions with the KasA protein that we descri<em>b</em>e may <em>b</em>e applied to evolve a next-generation therapeutic strategy for tu<em>b</em>erculosis (TB).(<em>b</em>)IMPORTANCE</<em>b</em>) Cell wall <em>b</em>iosynthesis inhi<em>b</em>itors have proven highly effective for treating tu<em>b</em>erculosis (TB). We discovered and validated mem<em>b</em>ers of the indazole sulfonamide class of small molecules as inhi<em>b</em>itors of <i>Myco<em>b</em>acterium tu<em>b</em>erculosis</i> KasA-a key component for <em>b</em>iosynthesis of the mycolic acid layer of the <em>b</em>acterium's cell wall and the same pathway as that inhi<em>b</em>ited <em>b</em>y the first-line antitu<em>b</em>ercular drug isoniazid (<em>INH</em>). One lead compound, DG167, demonstrated synergistic lethality in com<em>b</em>ination with <em>INH</em> and a transcriptional pattern consistent with <em>b</em>actericidality and loss of persisters. Our results also detail a novel dual-<em>b</em>inding mechanism for this compound as well as su<em>b</em>stantial structure-activity relationships (SAR) that may help in lead optimization activities. Together, these results suggest that KasA inhi<em>b</em>ition, specifically, that shown <em>b</em>y the DG167 series, may <em>b</em>e developed into a potent therapy that can synergize with existing antitu<em>b</em>erculars.

<A<em>b</em>stractText>Women with hereditary angioedema due to C1-inhi<em>b</em>itor deficiency (HAE-C1<em>INH</em>) experience more frequent and severe angioedema attacks compared with men. Fluctuations in female sex hormones can influence HAE attack frequency and severity. Su<em>b</em>cutaneous C1-<em>INH</em> (C1-<em>INH</em> [SC]) is indicated as routine prophylaxis to prevent HAE attacks. In this post hoc su<em>b</em>group analysis, we evaluated the efficacy and safety of C1-<em>INH</em> (SC) in female su<em>b</em>jects with HAE-C1<em>INH</em> enrolled in an open-la<em>b</em>el extension of the pivotal phase III COMPACT trial.</A<em>b</em>stractText><A<em>b</em>stractText>In this multicenter, randomized, parallel-arm trial, eligi<em>b</em>le su<em>b</em>jects (age ≥ 6 years with ≥ 4 attacks over 2 consecutive months) received C1-<em>INH</em> (SC) 40 IU/kg or 60 IU/kg twice weekly for 52 to 140 weeks. Analyses of efficacy endpoints were performed for all female su<em>b</em>jects and those of child<em>b</em>earing age (age ≥ 15 to ≤ 45 years), including su<em>b</em>jects who <em>b</em>ecame pregnant during the evaluation period.</A<em>b</em>stractText><A<em>b</em>stractText>Overall, 91% (69/76) of female su<em>b</em>jects were classified as responders (≥ 50% reduction in HAE attacks relative to the pre-study period); 82% experienced < 1 attack/4 weeks. The median num<em>b</em>er of attacks/month was 0.10, with 96% median reduction in attacks relative to the pre-study period. Results were similar in the su<em>b</em>group of su<em>b</em>jects of child<em>b</em>earing age. Four women who <em>b</em>ecame pregnant during the trial and were exposed to C1-<em>INH</em> (SC) during the first trimester delivered healthy <em>b</em>a<em>b</em>ies with no congenital a<em>b</em>normalities.</A<em>b</em>stractText><p><div>(<em>b</em>)Conclusions</<em>b</em>)</div>C1-<em>INH</em> (SC) prophylaxis was safe and effective in women with HAE-C1<em>INH</em>, including those of child<em>b</em>earing age. Four women exposed to C1-<em>INH</em> (SC) during the first trimester had uneventful pregnancies and delivered healthy <em>b</em>a<em>b</em>ies.<i>Trial registration</i> Clinicaltrials.gov identifier NCT02316353 (Registered Decem<em>b</em>er 10, 2014); https://clinicaltrials.gov/ct2/show/NCT02316353.</p>

<A<em>b</em>stractText>Acute liver injury (ALI) is associated with the oxidative stress and apoptosis in liver. Recent studies have shown that miR-195, a critical mem<em>b</em>er of miR-15 family, has modulated the apoptosis in various organic diseases. However, it is elusive whether miR-195 regulation exert a hepatic ameliorative effect on ALI <em>b</em>y the suppression of apoptosis and oxidative stress levels. We aimed to explore the regulated role of miR-195 in acute liver injury via the current study.</A<em>b</em>stractText><p><div>(<em>b</em>)METHODS</<em>b</em>)</div>C57BL/6 J mice (male, seven-week, 18-20 g) were administrated intraperitoneal injection with tetrachloromethane (CCl<su<em>b</em>)4</su<em>b</em>)) to induce ALI. miR-195 inhi<em>b</em>itor or mimics loaded in lentivirus vectors (miR-195 <em>INH</em> or MMC) and Pim-1 loaded in Adeno-associated viral vectors (AAV-Pim-1) were respectively delivered into mouse tail intravenous to esta<em>b</em>lish silence or overexpression of miR-195 and overexpression of Pim-1. Western <em>b</em>lotting, Reverse Transcription-Polymerase Chain Reaction (RT-PCR), enzyme linked immunosor<em>b</em>ent assay (ELISA) technique, Immunohistochemistry (IHC) and Hematoxylin-eosin (H&E) staining were conducted to measure miR-195 and Pim-1 expression, apoptosis and oxidative stress levels, histological and functional change.</p><A<em>b</em>stractText>We found that the expression of miR-195 markedly increased in CCl4-induced ALI. Besides, we demonstrated that the silence of miR-195 attenuated the apoptosis and oxidative stress via up-regulating Pim-1 in CCl4-induced ALI. Moreover, the inhi<em>b</em>ition of miR-195 protected the integrity and function of liver tissue.</A<em>b</em>stractText><A<em>b</em>stractText>The a<em>b</em>ove results showed that the suppression of miR-195 ameliorated ALI through inhi<em>b</em>iting apoptosis and oxidative stress via targeting Pim-1. Our research provided a novel scheme that the miR-195 modulation in process of ALI may <em>b</em>e an effective therapy method and verifies a promising target for diagnostic and therapeutic strategy of miRNAs.</A<em>b</em>stractText>

Kinins (peptides related to bradykinin, BK) are formed from circulating substrates, the kininogens, by the action of two proteases, the kallikreins. The only clinical application of a BK receptor ligand, the B₂ receptor antagonist icatibant, is the treatment of the rare hereditary angioedema (HAE) caused by the deficiency of C1-esterase inhibitor (C1-INH). Less common forms of HAE (genetic variants of factor XII, plasminogen, kininogen) are presumably mediated by increased BK formation. Acquired forms of BK-mediated angioedema, such as that associated with angiotensin-I converting enzyme (ACE) inhibition, are also known. Antibody-based analytical techniques are briefly reviewed, and support that kinins are extremely short-lived, prominently cleared by ACE. Despite evidence of continuous activation of the kallikrein-kinin system in HAE, patients are not symptomatic most of the time and their blood or plasma obtained during remission does not generate excessive immunoreactive BK (iBK), suggesting effective homeostatic mechanisms. HAE-C1-INH and HAE-FXII plasmas are both hyperresponsive to fibrinolysis activation. On another hand, we suggested a role for the alternate tissue kallikrein-kinin system in patients with a plasminogen mutation. The role of the BK B₁ receptor is still uncertain in angioedema states. iBK profiles under in vitro stimulation provide fresh insight into the physiopathology of angioedema.

Keywords: acquired angioedema; analytical techniques for kinins; bradykinin; hereditary angioedema; kallikrein–kinin system.