Indole chalcones were designed and synthesized as a promising set of compounds against H₃₇Rv strain of Mycobacterium tuberculosis. Within this library of compounds, (E)-1-(furan-3-yl)-3-(1H-indol-3-yl)prop-2-en-1-one (18), (E)-3-(1H-indol-3-yl)-1-(thiophen-2-yl)prop-2-en-1-one (20) and (E)-2-((1H-indol-2-yl)methylene)cyclopentan-1-one (24) displayed high anti-tubercular activity at 50 μg/ml with MIC values of 210, 197 and 236 μM respectively. The in-silico studies revealed that compound 18 exhibit binding modes similar to FAS-II inhibitors like INH or Thiolactomycin against KasA protein. Cytotoxicity assay results suggest that the compounds 18, 20 and 24 are non-cytotoxic to human megakaryocytes and murine B cells.

The binding reactions of the inhibitor drugs, SB 203580, SKF 86002, and p38 INH.1 to the isoforms 1 and 2 splice variants of p38alpha MAP kinase and their C162S mutants, as determined from ITC measurements from 25 to 35 degrees C, are totally enthalpically driven with binding constants ranging from 10(7)M(-1) for SKF 86002 and SB 203580 to 10(9)M(-1) for p38 INH.1. Interactions of p38 INH.1 with an additional hydrophobic pocket of the kinase would account for its large increase in K(b). DSC scans exhibited single unfolding transitions for the isoforms, their mutants, and the mutants bound to the drug inhibitors. Two transitions, however, were observed for the isoform-drug complexes of SB 203580 and p38 INH.1 and were attributed to decoupled unfolding of the N- and C-terminal domains of the kinase. The C-terminal domain of isoform 1 is estimated to be less stable than of isoform 2 by 15 kJ mol(-1).

OBJECTIVE

To investigate the changes of serum inhibin (Inh)-A, Inh-B concentrations during menopausal transition and the time relationship between changes of serum Inh-A, Inh-B and other reproductive hormone levels.

METHODS

Serum Inh-A, Inh-B concentrations were measured by Serotec modified two-site enzyme immunoassay during different phases of normal menstrual cycle in 10 healthy reproductive women, any time of 10 postmenopausal women. So did the serum Inh-A on 5 - 9 days prior to next period (premenstrual phase) and Inh-B determinations on the 3rd day of menstrual cycle in 40 women of age 43 - 52 (menopausal transition group). In addition, serum follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E(2)), progesterone (P) levels were also determined when necessary for the purpose of analysis.

RESULTS

The fluctuation patterns of serum Inh-A, Inh-B concentrations during normal menstrual cycle were completely different. About 48% of women during menopausal transition had normal luteal function as shown by the P levels during the premenstrual phase. The only significant change in these women as compared with the young was decrease of Inh-A concentration [(24.7 +/- 13.0) ng/L Vs (42.9 +/- 12.1) ng/L, P = 0.017] in the same phase. Further significant declines of serum Inh-A levels were seen in the luteal phase defect (LPD) and anovulatory (AOV) groups [(12.4 +/- 10.2) ng/L and (5.3 +/- 3.8) ng/L, P = 0.033, P < 0.000 1 respectively], until undetectable in the postmenopausal group. The day 3 Inh-B levels tended to decrease in the normal luteal and LPD groups, became undetectable in the AOV and postmenopausal groups (P = 0.001). Day 3 Inh-B levels was significantly lower in women with day 3 FSH>> or = 10 IU/L than those with < 10 IU/L [(16.2 +/- 4.0) ng/L Vs (62.0 +/- 43.8) ng/L]. The elevation of day 3 FSH, LH levels was not significant until the AOV group (P = 0.009, P = 0.027 respectively), and the drop of E(2) levels until the postmenopausal group (P < 0.001).

CONCLUSIONS

It is suggested that serum reproductive hormones should be measured in women of menopausal transition in order to know the stage of menopausal transition and to guide the clinical management. The decrease of serum Inh-A levels during the premenstrual phase is the earliest change of menopausal transition, and decrement of day 3 Inh-B levels a marker of decreased ovarian reserve.

BACKGROUND

Successful structural characterization of glycans often requires derivatization prior to mass spectrometric analysis. Here we report on a new derivatization reagent for glycans, biotinylated isonicotinic hydrazide, allowing glycan analysis by both mass spectrometry (MS) and biochemically. Fragmentation behavior in MS and its use in structural elucidation were investigated and compared with other labels.

METHODS

Glycans, released from ribonuclease B and ovalbumin, were derivatized with hydrazine labels (isoniazid (INH), biotinylated isonicotinic hydrazide (BINH) and biotinamidocaproylhydrazide (BACH)). In addition, native counterparts and 2-aminobenzamide (2-AB) derivatives were prepared. Comparative matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI TOF/TOF) experiments were carried out to investigate the fragmentation pattern of the derivatives. Finally, the capability of BINH derivatives to bind lectins was explored.

RESULTS

Generally, derivatization provided beneficial enhancement in the mass spectrometric signal intensity as compared to native counterparts. The mass spectrometric fragmentation varied with the kind of label used. The most significant structure-revealing ions (cross-ring cleavages) were observed in the spectra of BINH derivatives, whereas mainly glycosidic cleavages were found with native form of glycans and 2-AB derivatives.

CONCLUSIONS

Hydrazine derivatization provided the means to obtain structurally informative fragment ions. Due to BINH derivatization, specific fragments of the isomers allowed the identification of diverse glycans. The derivatization reaction can be carried out without the need for purification. The biotin residue of BINH enabled for biochemical studies, i.e. protein-glycan interactions.

(<em>b</em>)O<em>b</em>jective:</<em>b</em>) Possi<em>b</em>le effects of the vagus inhi<em>b</em>ition and stimulation on the hypothalamic nuclei, myenteric plexes and the vagus nerve were investigated. (<em>b</em>)Methods:</<em>b</em>) The female rats divided to the inhi<em>b</em>ition (<em>INH</em>), stimulation (STI) and, sham (SHAM) groups were fed with high fat diet (including 40% of energy from animal fat). After nine weeks, the rats were allowed to recover for 4 weeks in <em>INH</em> group. In STI group, the left vagus nerve stimulated (30 Hz/500 msn/30 sec.) starting 2nd post operative day for 5 minutes during 4 weeks. Healthy female rats used as control (CONT). Then, tissue samples were analyzed <em>b</em>y <em>b</em>iochemical, histological and stereological methods. (<em>b</em>)Results:</<em>b</em>) The mean num<em>b</em>er of the neurons in the arcuate nucleus of the <em>INH</em> group was significantly less; <em>b</em>ut, that is significantly more in the STI group compared to the other groups. The neuronal density of ventromedial nucleus in the STI group was higher; while the density in the <em>INH</em> group was lower than the other groups. In the dorsomedial nucleus, neuron density of the <em>INH</em> group was lower than the other groups. In terms of the myenteric plexus volumes, that of the <em>INH</em> group was lowest. The myelinated axon num<em>b</em>er in the <em>INH</em> group was significantly highest. The myelin sheath thickness and axon area of the <em>INH</em> group was significantly lower than the other groups. (<em>b</em>)Discussion:</<em>b</em>) The results of the study show that the vagal inhi<em>b</em>ition is more effective than the vagal stimulation on the weight loss in the o<em>b</em>esity.

Keywords: NPY; Obesity; POMC; hypothalamic nuclei; myenteric plexus; vagal inhibition; vagal stimulation.

C1-inhibitor (C1-Inh) is a serine esterase proteinase inhibitor (serpin) which plays an important role in regulating serine proteinases of the early inflammatory response. In this study, we describe a novel and versatile polyclonal antibody capture assay to examine C1-Inh consumption in vivo. This assay has advantages over previously described methods of measuring C1-Inh consumption as it allows the assessment of the relative amounts of native, complexed and cleaved inhibitor circulating in plasma. By using polyclonal antibodies specific for other complement proteins, the C1-Inh capture assay was adapted to measure in vivo activation of C3, C4 and factor B. C1-Inh consumption and complement activation were examined in the plasma of 21 normal individuals, 24 individuals with systemic lupus erythematosus (SLE), nine individuals with adult respiratory distress syndrome (ARDS) and in the paired plasma and synovial fluid from 18 patients with rheumatoid arthritis (RA). The C1-Inh capture assay revealed native, cleaved and complexed C1-Inh migrating at 115 kDa, 96 kDa and 209-225 kDa respectively, in normal plasma. C1-Inh consumption was increased in the plasma of all the inflammatory disorders examined, in comparison to normal plasma. It is proposed that this serpin capture assay could be adapted to the study of serpin involvement in a wide variety of inflammatory disorders.

Novel 1,1'-(5,5'-(1,4-phenylene)bis(3-aryl-1H-pyrazole-5,1-(4H,5H)-diyl))diethanones 7-12 were tested for their antimicrobial activity by disc diffusion and twofold serial dilution method against the tested bacterial and fungal strains. Compounds 7 against Micrococcus luteus, 8 against β-Heamolytic streptococcus, M. luteus, Klebsiella pneumonia, Microsporum gypseum, 9 against Staphylococcus aureus, Shigella flexneri, Vibreo cholerae, Pseudomonas aeruginosa, Aspergillus flavus, Mucor indicus, 10 against Salmonella typhii, S. flexneri, M. gypseum, 11 against K. pneumonia, M. gypseum, 12 against K. pneumonia, and M. gypseum show superior zone of inhibitions and exhibited excellent antibacterial and antifungal activities at a MIC value of 6.25 μg/mL. Moreover, all the tested compounds 7-12 revealed promising antitubercular activity against Mycobacterium tuberculosis H37Rv and INH-resistant M. tuberculosis. Compounds 8 against M. tuberculosis and 11 against INH-resistant M. tuberculosis exhibited the percentage of reduction in RLU at 89 and 85%, respectively.

<A<em>b</em>stractText>To screen new serum meta<em>b</em>olic <em>b</em>iomarkers for different drug resistance profiles of pulmonary tu<em>b</em>erculosis (TB) and explore their mechanisms and functions.</A<em>b</em>stractText><A<em>b</em>stractText>We collected serum samples from TB patients with drug sensitivity (DS), monoresistance to isoniazid (MR-<em>INH</em>), monoresistance to rifampin (MR-RFP), multidrug resistance (MDR), and polyresistance (PR). The meta<em>b</em>olites in the serum samples were extracted <em>b</em>y oscillatory and deproteinization for LC-MS/MS analysis, and the results were normalized <em>b</em>y Pareto-scaling method and analyzed using Meta<em>b</em>oanalyst 4.0 software to identify the differential meta<em>b</em>olites. The differential meta<em>b</em>olites were characterized <em>b</em>y function enrichment and co-expression analysis to explore their function and possi<em>b</em>le pathological mechanisms.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>Compared with the DS group, 286 a<em>b</em>normally expressed meta<em>b</em>olites were identified in MR-<em>INH</em> group, 362 in MR-RPF group, 277 in MDR group and 1208 in PR group <em>b</em>y LC-MS/MS analysis. Acetylagmatine (<i>P</i> < 0.05), aminopentol (<i>P</i> < 0.05), and tetracosanyl oleate (<i>P</i> < 0.05) in MR-<em>INH</em> group; Ala His Pro Thr (<i>P</i> < 0.001) and glycinoprenol-9 (<i>P</i> < 0.05) in MR-RFP group; trimethylamine (<i>P</i> < 0.05), penaresidin A (<i>P</i> < 0.05), and verazine (<i>P</i> < 0.05) in MDR group; and PIP (18:1(11Z)/ 18:3(6Z, 9Z, 12Z)) (<i>P</i> < 0.001), Pro Arg Trp Tyr (<i>P</i> < 0.001), N-methyldioctylamine (<i>P</i> < 0.001), and phytolaccoside E (<i>P</i> < 0.05) in PR group all showed significant differential expressions. Significant differential expressions of phthalic acid mono-2-ethylhexyl ester (<i>P</i> < 0.05) and eicosanoyl-EA (<i>P</i> < 0.05) were found in all the drug resistant groups as compared with DS group.</p><A<em>b</em>stractText>Acetylagmatine, aminopentol, tetracosanyl oleate, Ala His Pro Thr, glycinoprenol-9, trimethylamine, penaresidin A, verazine, PIP(18:1(11Z)/18:3(6Z, 9Z, 12Z)), Pro Arg Trp Tyr, N-methyldioctylamine, phytolaccoside E, phthalic acid mono-2-ethylhexyl ester, and eicosanoyl-EA are potentially new <em>b</em>iomarkers that indicate monoresistance, multi-drug resistance and polyresistance of Myco<em>b</em>acterium tu<em>b</em>erculosis. The com<em>b</em>ined use of these <em>b</em>iomarkers potentially allows for assessment of drug resistance in TB and enhances the diagnostic sensitivity and specificity.</A<em>b</em>stractText>

Acquired deficiency in C1 esterase inhibitor (C1 INH) was first described by Caldwell in 1972. Since that date, about 30 cases have been reported, in most cases during proliferative lymphocyte B syndromes. The acquired C1 INH deficiency can provoke episodes of angioneurotic edema as in hereditary AE. The complement profile differs, notably by the usual sudden fall in C1. Documented data suggest consumption of complement and therefore of C1 INH. The present case is the first reported of an acquired C1 INH deficiency during a cold hemagglutinin disease. The activation of complement by the classical pathway appears provoked by tumoral cells rather than a humoral factor, as suggested by the efficacy of anti-tumoral therapy in contrast to plasmapheresis in the present case. A possible mechanism for the C1 INH deficiency is discussed.

<A<em>b</em>stractText>To investigate the effect of autophagy-related gene (ATG) on the drug resistance of myco<em>b</em>acteria <em>b</em>y regulating autophagy.</A<em>b</em>stractText><A<em>b</em>stractText>In the present study, primary macrophages were selected as o<em>b</em>jects of study. The cell lines with ATG13 and ATG6 interference and sta<em>b</em>le overexpression were constructed with Crisp/Case technique and verified <em>b</em>y fluorescence quantitative polymerase chain reaction (PCR) and western <em>b</em>lotting, and the qualified cells were used for su<em>b</em>sequent experiments. Then the a<em>b</em>ove different mutant and wild-type cells were cultured in Dul<em>b</em>ecco's Modified Eagle medium (DMEM) containing fetal <em>b</em>ovine serum for 5 h, and Myco<em>b</em>acterium tu<em>b</em>erculosis H37Rv was added, followed <em>b</em>y co-culture for 4 h. The cells were treated and co-cultured with isoniazid (<em>INH</em>, 0.05 mg/L), rifampicin (RFP, 0.4 mg/L) and etham<em>b</em>utol (EMB, 25 mg/L) for 3 d. Then the cells were sampled and stained with monodansylcadaverine (MDC), and autophagy was o<em>b</em>served. Finally, an appropriate num<em>b</em>er of cells were taken and cultured in the modified L-G medium, and the <em>b</em>acteria were counted.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>The results of fluorescence quantitative PCR and western <em>b</em>lotting revealed that the messenger ri<em>b</em>onucleic acid (mRNA) transcription levels and protein expression levels of ATG13 and ATG6 in cells significantly declined after using Crisp/Case. The MDC staining showed that ATG13 and ATG6 interference could significantly reduce the num<em>b</em>er of autophagosomes in cells, while ATG13 and ATG6 overexpression could significantly increase the num<em>b</em>er of autophagosomes in cells. Compared with wild-type cells, the num<em>b</em>er of myco<em>b</em>acteria was o<em>b</em>viously increased in myco<em>b</em>acterium-infected cells with ATG13 and ATG6 interference after they were treated with <em>INH</em>, RFP and EMB, displaying a significant difference (<i>P</i><0.05), while the num<em>b</em>er of myco<em>b</em>acteria was o<em>b</em>viously decreased in myco<em>b</em>acterium-infected cells with ATG13 and ATG6 overexpression after they were treated with <em>INH</em>, RFP and EMB, also a significant difference (<i>P</i><0.05).</p><A<em>b</em>stractText>ATG and other autophagy-related genes can affect the drug resistance of myco<em>b</em>acteria through regulating autophagy.</A<em>b</em>stractText>

<p><div>(<em>b</em>)INTRODUCTION</<em>b</em>)</div>HIV-exposed uninfected (HEU) infants in tu<em>b</em>erculosis (TB) endemic settings are at high risk of <i>Myco<em>b</em>acterium tu<em>b</em>erculosis</i> (Mt<em>b</em>) infection and TB disease, even in the a<em>b</em>sence of known Mt<em>b</em> exposure. Because infancy is a time of rapid progression from primary infection to active TB disease, it is important to define when and how TB preventive interventions exert their effect in order to develop effective prevention strategies in this high-risk population.</p><A<em>b</em>stractText>We designed a non-<em>b</em>linded randomised controlled trial to determine efficacy of isoniazid (<em>INH</em>) to prevent primary Mt<em>b</em> infection among HEU children. Target sample size is 300 (150 infants in each arm). Children are enrolled at 6 weeks of age from maternal and child health clinics in Kenya and are randomised to receive 12 months of daily <em>INH</em> ~10 mg/kg plus pyridoxine or no <em>INH</em>. The primary endpoint is Mt<em>b</em> infection, assessed <em>b</em>y interferon-gamma release assay QuantiFERON-TB Gold Plus (QFT-Plus) or tu<em>b</em>erculin skin test after 12 months post-enrolment. Secondary outcomes include severe adverse events, expanded Mt<em>b</em> infection definition using additional QFT-Plus supernatant markers and determining correlates of Mt<em>b</em> infection. Exploratory analyses include a com<em>b</em>ined outcome of TB infection, disease and mortality, and sensitivity analyses excluding infants with <em>b</em>aseline TB-specific responses on flow cytometry.</A<em>b</em>stractText><A<em>b</em>stractText>An external and independent Data and Safety Monitoring Board monitors adverse events. Results will <em>b</em>e disseminated through peer-reviewed journals, presentations at local and international conferences to national and glo<em>b</em>al policy-makers, the local community and participants.</A<em>b</em>stractText><A<em>b</em>stractText>NCT02613169; Pre-results.</A<em>b</em>stractText>

The antimicrobial susceptibility testing for Mycobacterium tuberculosis by the bioluminescence assay of adenosine triphosphate(ATP) derived from living mycobacteria was improved introducing filamentous cell treatment(FCT) reported for beta-lactam susceptibility test of Pseudomonas aeruginosa by Hattori. Before ATP extraction, bacterial cells were treated with the FCT reagent for 30 minutes at room temperature. Adenosine phosphate deaminase in the FCT reagent simultaneously digested the extracted ATP and released ATP in a liquid culture of M. tuberculosis H37Rv and the RLU level was decreased markedly. Using this improved ATP method, we determined the ATP contents of M. tuberculosis inoculated into Middle-brook 7H9 broth medium with or without drugs. In ethambutol(EB) susceptibility, the ATP method reported previously, showed false-resistance when judged within 7 days. To eliminate false-resistance in EB susceptibility we applied the modified ATP method with FCT treatment to strains determined EB susceptible by reference methods. Using this modified ATP method, we could judge EB susceptibility of 5 ATCC reference strains within 3 days, and these of 15 clinical isolates of M. tuberculosis within 5 days. And all the results obtained were coincident between the ATP method and the reference methods. The reproducibility of this modified ATP method was evaluated with six ATCC reference strains at the concentrations of 0.1 microgram/ml of isoniazid(INH), 2.0 micrograms/ml of rifampicin(RFP), 2.5 micrograms/ml of EB, 2.0 micrograms/ml of streptomycin(SM), and 5.0 micrograms/ml of kanamycin(KM). The test was repeated six times. Reduction of ATP contents were observed in susceptible strains but not in resistant ones within 3 days of cultivation and susceptibilities to drugs could be determined within 3 days at every time when combined FCT to the ATP method. And highly reproducible results were obtained. It is strongly suggested that this modified method is simple, rapid, highly reproducible and nonradiometric, and could be used for the assessment of drug susceptibility for M. tuberculosis.

BACKGROUND

Current international guidelines recommend 6-9 months of isoniazid (INH) preventive chemotherapy to prevent the development of active tuberculosis (TB) in susceptible children exposed to Mycobacterium tuberculosis. However, this is dependent on good adherence, as shown by previous studies.

OBJECTIVE

This study was conducted to describe the outcome of screening of contact children aged 5 years or less with household exposure to an adult pulmonary TB index case to determine the prevalence and possible risk factors of infection among contact children and to determine the extent and outcome of adherence of contact children to unsupervised INH chemoprophylaxis for 6 months.

METHODS

A descriptive facility-based cross-sectional study was conducted from March 2009 to August 2010. Research settings were three of the National TB control program chest dispensaries (primary care facilities) in Alexandria, Egypt. Facility-based TB treatment registers of the previous 3 months were used to identify all new adult pulmonary TB cases. All children aged 5 years or less living in the same house as the index cases were identified and screened for TB. The contact children were given unsupervised INH preventive chemotherapy once active TB was excluded. Adherence to and outcome of preventive chemotherapy were followed up. Preventive chemotherapy consisted of unsupervised INH monotherapy for 6 months with monthly collection of tablets from the clinic. Adherence was documented after completion of the 6-month preventive treatment period. Adherence was considered reasonable if tablets were collected for more than 4 months, poor if collected for 2-4 months, and very poor if collected for less than 2 months.

METHODS

(a) Prevalence of infection and disease and the possible risk factors among contacts. (b) The extent and outcome of adherence to unsupervised INH chemoprophylaxis among contact children. (c) Factors behind poor adherence.

RESULTS

In total, 197 adult TB index cases from 187 households were identified. In all, 297 children aged 5 years or less experienced household exposure, of whom 252 (84.9%) were fully evaluated. Tuberculin test was positive in 136 of the 252 child contacts (54.0%), of whom 130 were contacts of sputum-positive patients and six were contacts of sputum-negative patients. The important risk factors for transmission of TB infection were younger age, male sex, severe malnutrition, absence of BCG vaccination, contact with a sputum-positive adult who was a source case, household overcrowding, and exposure to environmental tobacco smoke. Thirty-three children were diagnosed and treated for TB at the baseline screening and 217 received preventive INH chemotherapy. Of the children who received preventive chemotherapy, only 36 (16.6%) completed at least 4 months of unsupervised INH monotherapy. During the subsequent follow-up period, eight children developed TB (secondary attack rate for TB disease was 3.7%), of whom four received no preventive chemotherapy and four were poorly adherent.

CONCLUSIONS

The prevalence of TB infection and clinical disease among children in household contact with adult patients is high, and risk is significantly increased because of child contact, index patients and environmental factors. Adherence to 6 months of unsupervised INH chemoprophylaxis was very poor.

BACKGROUND

Toll-like receptor 4 (TLR4), an innate immune receptor, is suspected to play a key role in the postprandial inflammation that is induced by a high-fat meal rich in saturated fatty acids (SFA). Our objective was to test this hypothesis by using a specific competitive inhibitor of TLR4 (INH) vs vehicle (VEH) administered immediately before a high-SFA meal in rats.

METHODS

First, in a cross-over kinetic study of 12 rats receiving INH and VEH i.v. 10 min before the test meal, we measured plasma inflammatory and vascular markers for 6 h. Then, in 20 rats, 3 h after INH or VEH followed by the test meal (parallel study), we measured the mRNA level of a set of cytokines (Il1-β, Il-6, Tnfα, Mcp-1, Pai-1), and of Tlr4 and Tlr2 in the adipose tissue and the liver, and that of adhesion molecules (Icam-1 and Vcam-1) in the aorta.

RESULTS

Plasma IL-6 and PAI-1 increased >4-fold at 3-4 h after test-meals, very similarly after INH as compared to VEH. The expression of TLR2 and of all measured cytokine genes in the adipose tissue was dramatically higher after INH (vs VEH). In the liver, gene expression of Il1-β, Tnfα, Mcp-1 and Tlr2, was also higher after INH, though more moderately, whereas that of Il-6 and Pai-1 was similar between groups. INH did not affect mRNA level of Icam-1 and Vcam-1 in the aorta.

CONCLUSIONS

TLR4 activation is not specifically required to mediate systemic postprandial inflammation and we propose that TLR2 and TLR4 exert a dual and interdependent mediation of the postprandial inflammatory response, at least in the adipose tissue.

Because recent investigations showed that the use of isoniazid (INH) severely impaired the local immune reaction to intravesical bacillus Calmette-Guérin (BCG) in the bladder of guinea pigs, in this study the effect of INH in man has been investigated. Patients were treated with BCG with or without oral INH. The concentration of free INH in most urine samples of patients treated with BCG/INH was much higher (mean 38.0 +/- 60.9 micrograms INH/ml) than the minimal inhibitory concentration (MIC; 0.1 microgram INH/ml), suggesting at least a bacteriostatic potential of the INH present. However, in vitro studies showed that these urinary concentrations of INH did not kill BCG organisms effectively, even at a concentration of 150 micrograms/ml for 24 h. After the fifth and sixth BCG instillations a significant increase in the concentration of cytokines (IL2, IL6, IL8 and TNFa), IgG and IgA antibodies to BCG and the number of leukocytes in urine was observed. The leukocytes mainly consisted of granulocytes, besides monocytes/macrophages and, in lower amounts, T- and B-lymphocytes and natural killer (NK) cells. The absolute number of granulocytes and the concentration of IgG antibodies after BCG instillation were significantly suppressed by INH, whereas INH appeared to have no effect on the urinary cytokine and IgA antibody concentrations or the total number and phenotype of the leukocytes present. In conclusion, the results of this study indicate that INH does not impair the local immunological stimulation after BCG instillation in man as severely as was observed in the guinea pig and it may be expected that INH does not impair the antitumor efficacy of BCG.

Aim(s): The present study was conducted to formulate and investigate liposomes for the dual drug delivery based on anti-tubercular drug(s) combination i.e., Isoniazid (INH) and Rifampicin (RIF).

Materials & methods: Mannosylated and non mannosylated liposomes were prepared by lipid thin film hydration method, using DSPC: Chol at a molar ratio 6:4 while in case of mannosylated liposomes DSPC: Chol: Man-C4-Chol at a molar ratio 6.0:3.5:0.5 were used and extensively characterized. The particle size and zeta potential were recorded to be 1.29 ± 0.24µm and -9.1 ± 0.11mV. The drug entrapment (%) was recorded to be 84.7 ± 1.25% for Rifampicin and 31.8 ± 0.12% for Isoniazid.

Results: The antitubercular activity studied in Balb/C mice was maximum in the case of mannosylated liposomes. The biodistribution studies also revealed higher drug(s) concentration (accumulation) maintained over a protracted period.

Conclusions: The liposomal preparations are passively as well as actively uptaken by the alveolar macrophages which are the cellular tropics of infection. The mannosylated liposomes appear to be a potential carrier for dual drug delivery and targeted antitubercular therapy.

Keywords: active targeting; combination drug therapy; infectious disease; macrophages; mannosylated liposomes; phagocytosis; – Mycobacterium tuberculosis.

(<em>b</em>)OBJECTIVE:</<em>b</em>) 1) To determine the prevalence of dia<em>b</em>etes mellitus and impaired fasting glucose (IFG) in patients with TB and HIV co-infection, and 2) to investigate the effect of fasting plasma glucose (FPG) on rifampicin (RIF) and isoniazid (<em>INH</em>) serum concentrations.(<em>b</em>)DESIGN:</<em>b</em>) Retrospective data analysis of a cohort of HIV-infected adults with newly diagnosed pulmonary TB. Plasma glucose and TB drug levels were o<em>b</em>tained at Week 0, 2, 8 and 24 of TB treatment.(<em>b</em>)RESULTS:</<em>b</em>) A total of 107 patients were included in this analysis. Random plasma glucose ≥200 mg/dL was found in 1/53 (2%) participant at Week 0. The prevalence of FPG ≥ 126 mg/dL decreased from 8/41 (20%) at Week 2 to 3/89 (3%) at Week 24. IFG (100-125 mg/dL) was o<em>b</em>served in 23/41 (56%) participants at Week 2, and 39/89 (44%) at Week 24. FPG was inversely correlated with lower area under the curve (AUC<su<em>b</em>)0-24h</su<em>b</em>)) for RIF (c = -0.52; 95%CI -0.84 to -0.21; <i>P</i> = 0.001). FPG was not associated with lower <em>INH</em> AUC<su<em>b</em>)0-24h</su<em>b</em>).(<em>b</em>)CONCLUSION:</<em>b</em>) We found a high prevalence of FPG ≥ 126 mg/dL, which decreased significantly during treatment, and a high proportion of IFG at the end of TB treatment. Higher FPG was associated with lower AUC for RIF.

In recent years, flexible and sensitive pressure sensors are of extensive interest in healthcare monitoring, artificial intelligence and national security. In this context, we report the synthetic procedure of a 3D metal-organic framework (MOF) comprising with cadmium (Cd) metals as nodes and isoniazid (INH) moieties as organic linkers (CdI2-INH=CMe2) for designing self-polarized ferroelectret based highly mechano-sensitive skin sensor. The as synthesized MOF preferentially nucleate stable piezoelectric β-phase in poly(vinylidene fluoride) (PVDF) and also give rise to a porous ferroelectret composite film. Benefiting from the porous structure of 3D MOF, composite ferroelectret film based ultra-sensitive pressure sensor (mechano-sensitivity of 8.52 V/kPa within 1kPa of pressure range) as well as high throughput (e.g., power density of 32 W/cm2) mechanical energy harvester (MEH) has been designed. Simulation based finite element method (FEM) analysis indicating that geometrical stress confinement effect within the inter-pore region of the ferroelectret structure synergistically influences the mechano-electrical property of the MEH. In addition, 143 pC/N (~ 4.5 times higher than commercial piezoelectric PVDF film) of piezoelectric charge coefficient (d33) magnitude and superior response time (tr~8 ms) of this composite ferroelectret film enable to detect different physiological signals such as coughing, pronunciation, gulping behaviour makes it a promising candidate for early intervention of healthcare which may play a significant role in accurate alert of influenza and chronic obstructive pulmonary disease (COPD) related symptoms. In addition, MEH enables to track the subtle pressure change in wrist pulse that indicates its usefulness of effective mechano-sensitivity. Since the cardiovascular signal is one of the vital parameter that can determine the on-going physiological conditions, thus the wireless transmission of the detected wrist pulse signal has been demonstrated. All these features coupled with wireless data transmission indicate the promising application of MOF assisted composite ferroelectret film in non-invasive real time remote heath care monitoring.

(<em>b</em>)INTRODUCTION:</<em>b</em>) Testing for anti-TB drugs in small hair samples may serve as a non-invasive tool to measure cumulative drug exposure and/or adherence, as these determine treatment success. We aimed to assess how well hair assays of TB drugs predict TB treatment outcomes.(<em>b</em>)METHODS:</<em>b</em>) A small thatch of hair, ~30 strands, was cut from the occipital region in adults and children from a prospective TB cohort in India. Isoniazid (<em>INH</em>), acetyl-<em>INH</em> and pyrazinamide (PZA) were extracted from the hair samples and quantified using liquid-chromatography-tandem mass spectrometry. The relationship <em>b</em>etween drug concentrations in hair and time to unfavoura<em>b</em>le outcomes was assessed using Cox-proportional hazards regression models.(<em>b</em>)RESULTS:</<em>b</em>) A two-fold increase in hair acetyl-<em>INH</em> concentrations in the 264 participants in our cohort with hair assays for TB drugs indicated a lower hazard of unfavoura<em>b</em>le TB treatment outcomes (aHR 0.67, 95%CI 0.44-1.02) and TB treatment failure (aHR 0.65, 95%CI 0.42-1.01). Higher summed concentrations (a summed measure of <em>INH</em> and acetyl-<em>INH</em>) indicated a lower hazard of treatment failure (aHR 0.69, 95%CI 0.45-1.05)(<em>b</em>)CONCLUSION:</<em>b</em>) Hair levels of <em>INH</em> and its meta<em>b</em>olite may predict TB treatment outcomes, indicating the potential utility of this measure to assess and optimise TB treatment outcomes.

(<em>b</em>)Background:</<em>b</em>) Two sequential single-dose crossover dose-ranging studies were performed to evaluate the clinical efficacy and safety profile of epinephrine hydrofluroalkane (HFA) metered-dose <em>inh</em>aler (MDI) formulation at various doses in su<em>b</em>jects with asthma. (<em>b</em>)Methods:</<em>b</em>) In these multicenter, multiarm, dou<em>b</em>le-<em>b</em>linded, or evaluator-<em>b</em>linded studies, su<em>b</em>jects were randomized to receive the epinephrine HFA (Primatene^® MIST HFA) MDI medication at doses ranging from 90 to 440 μg/dose, as well as to a place<em>b</em>o (PLA) control and an active control of epinephrine CFC (chlorofluorocar<em>b</em>on) MDI (Primatene MIST CFC) at 220 μg/<em>inh</em>alation. (<em>b</em>)Results:</<em>b</em>) Spirometry testing for FEV1 (Forced Expiratory Volume in one second) demonstrated statistically significant improvements over PLA for epinephrine HFA MDI at all doses a<em>b</em>ove 125 μg, as the amount out of the actuator (i.e., mouthpiece). The efficacy results for epinephrine HFA MDI in the dose range of 125-250 μg were also compara<em>b</em>le to epinephrine CFC MDI (220 μg/<em>inh</em>). Safety assessments demonstrated minimal safety concerns for all treatment groups. No nota<em>b</em>le safety differences were o<em>b</em>served <em>b</em>etween the studied doses of epinephrine HFA MDI and the active control formulation of epinephrine CFC MDI. (<em>b</em>)Conclusion:</<em>b</em>) The findings indicate that epinephrine HFA MDI provided clinically significant <em>b</em>ronchodilator efficacy with minimal safety concerns in a dose range of 125-250 μg. These findings confirmed the optimal treatment doses of 125-250 μg that were appropriate for use in longer term 12 and 26 week chronic dosing studies of epinephrine HFA MDI for patients with intermittent or mild to moderate persistent asthma. Clinical trials registration num<em>b</em>er: NCT01025648.

Interstitial cells of Cajal, which express the calcium-activated chloride channel transmem<em>b</em>rane mem<em>b</em>er 16A (TMEM16A), are an important determinant of gastrointestinal (GI) motility. We previously identified the acylaminocycloalkylthiophene class of TMEM16A <em>inh</em>i<em>b</em>itors, which, following medicinal chemistry, gave analog 2-<em>b</em>romodifluoroacetylamino-5,6,7,8-tetrahydro-4<i>H</i>-cyclohepta[<i><em>b</em></i>]thiophene-3-car<em>b</em>oxylic acid <i>o</i>-tolylamide (TM<su<em>b</em>)<em>inh</em></su<em>b</em>)-23) with 30 nM half-maximal <em>inh</em>i<em>b</em>itory concentration. Here, we tested the efficacy of TM<su<em>b</em>)<em>inh</em></su<em>b</em>)-23 for <em>inh</em>i<em>b</em>ition of GI motility in mice. In isolated murine gastric antrum, TM<su<em>b</em>)<em>inh</em></su<em>b</em>)-23 strongly <em>inh</em>i<em>b</em>ited spontaneous and car<em>b</em>achol-stimulated rhythmic contractions. Pharmacokinetic analysis showed predicted therapeutic concentrations of TM<su<em>b</em>)<em>inh</em></su<em>b</em>)-23 for at least 4 h following a single oral or intraperitoneal dose at 10 mg/kg. Gastric emptying, as assessed following an oral <em>b</em>olus of phenol red or independently <em>b</em>y [^99mTc]-diethylenetriamine pentaacetic acid scintigraphy, was reduced <em>b</em>y TM<su<em>b</em>)<em>inh</em></su<em>b</em>)-23 <em>b</em>y ∼60% at 20 min. Interestingly, there was little effect of TM<su<em>b</em>)<em>inh</em></su<em>b</em>)-23 on <em>b</em>aseline whole-gut transit time or time to diarrhea induced <em>b</em>y castor oil. Consequent to the delay in gastric emptying, TM<su<em>b</em>)<em>inh</em></su<em>b</em>)-23 administration significantly reduced the elevation in <em>b</em>lood sugar in mice following an oral <em>b</em>ut not intraperitoneal glucose load. These results provide pharmacological evidence for involvement of TMEM16A in gastric emptying and suggest the utility of TMEM16A <em>inh</em>i<em>b</em>ition in disorders of accelerated gastric emptying, such as dumping syndrome, and potentially for improving glucose tolerance in dia<em>b</em>etes mellitus/meta<em>b</em>olic syndrome and enhancing satiety in o<em>b</em>esity.-Cil, O., Anderson, M. O., Yen, R., Kelleher, <em>B</em>., Huynh, T. L., Seo, Y., Nilsen, S. P., Turner, J. R., Verkman, A. S. Slowed gastric emptying and improved oral glucose tolerance produced <em>b</em>y a nanomolar-potency <em>inh</em>i<em>b</em>itor of calcium-activated chloride channel TMEM16A.

Co-administration of isonicotinic acid hydrazide (isoniazid, INH) and 17 alpha-ethinyl-17 beta-hydroxyestr-4-en-3-one (norethindrone, NE) resulted in the formation of the isonicotinyl hydrazone of norethindrone (INH-NE) in rat stomach. Rat liver metabolized the latter compound in vitro. The metabolic product was characterized, following its derivatization with p-methoxy-benzaldehyde (PMBA), by comparison of chromatographic and mass spectral properties with synthetic reference compound. Results showed that INH-NE was cleaved at the amide bond resulting in the formation of the hydrazone of norethindrone. The physicochemical characteristics of synthetic PMBA hydrazone of norethindrone are described.

Previous observations from our laboratory have demonstrated that the levels of immunoreactive inhibin (ir-inh) are elevated in almost all patients with granulosa cell tumors and in the majority of postmenopausal women with mucinous ovarian cancers. The present report confirms these findings in a larger group of post-menopausal women. Immunohistochemistry for the inhibin alpha. beta A and beta B sununits shows predominantly epithelial staining in granulosa cell tumors and in the majority of mucinous cancers. Serous cystadenocarcinomas also frequently show positive staining. Studies seeking to identify G alpha i-2 or FSH receptor mutations have provided negative results in contrast to other reports. Further studies of the roles of the inhibin-related family of peptides in ovarian cancer diagnosis and monitoring are clearly indicated.