Although color Doppler ultrasonography has been used to evaluate testicular blood flow in many species, very little has been done in goat. Eight male Shiba goats were exposed to a single intramuscular injection of either gonadotropin-releasing hormone (GnRH group; 1 µg/kg BW) or human chorionic gonadotropin (hCG group; 25 IU/kg BW). Plasma testosterone (T), estradiol (E2) and inhibin (INH) were measured just before (0 hr) and at different intervals post injection by radioimmunoassay. Testis volume (TV) and Doppler indices, such as resistive index (RI) and pulsatility index (PI) of the supratesticular artery, were measured by B-mode and color Doppler ultrasonography, respectively. The results indicated an increase in testicular blood flow in both groups, as RI and PI decreased significantly (P<0.05), but this increase was significant higher and earlier in hCG group (1 hr) than in the GnRH group (2 hr). A high correlation was found for RI and PI with both T (RI, r= -0.862; PI, r= -0.707) and INH in the GnRH group (RI, r=0.661; PI, r=0.701). However, a significant (P<0.05) correlation was found between E2 and both RI (r= -0.610) and PI (r= -0.763) in hCG group. In addition, TV significantly increased and was highly correlated with RI in both groups (GnRH, r= -0.718; hCG, r= -0.779). In conclusion, hCG and GnRH may improve testicular blood flow and TV in Shiba goats.

Three simple and sensitive methods for the assay of Nifurtimox (NIF) which is an active antitrypanocide were developed. These methods are based on the formation of coloured species by treating either its reduction product with 3-methyl-2-benzothiazolinone hydrazone (MBTH) in the presence of ferric chloride (method A) or its hydrolysis product with 2-thiobarbituric acid (TBA) (method B) or by oxidizing it with excess N-bromosuccinamide (NBS) and determining the consumed NBS using p-N-methylaminophenol sulphate (metol)-isonicotinic acid hydrazide (INH) (method C). All variables have been optimized and the reaction mechanisms presented. Regression analysis of Beer's plot showed good correlation in the concentration range of 2.5-10, 2.5-30 and 1.25-7.5 microg/ml for methods A, B and C, respectively. No interference was observed from the additives and the validity of the methods was tested by analysing the tablets. Recoveries were 99.2-100.9%.

The present paper deals with the preparation and characterization of a conjugate of isoniazid (INH) with the block copolymer methoxypoly(ethylene glycol)-b-poly(L-lysine) (mPEG-b-PLL). The structure of the conjugate (mPEG-b-PLL-INH) was verified by means of (1)H NMR, GPC, infrared spectroscopy, elemental analysis and powder X-ray diffraction. The conjugate contains six l-lysine units with five INH molecules, which are attached by means of pH-sensitive amidine bond. Under in vitro conditions, the conjugate is hydrolyzed and isoniazid is released (pH 4; 37 °C; t(1/2) ≈ 10 h).

The conformation of a hexapeptide sequence occurring in tropoelastin is discussed from the results obtained using a combined approach of theoretical conformational energy calculations on HCO-Val-Ala-Prb-Gly-OMe and 1h nmr studies on t-Boc-Val-Ala-Pro-Gly-Val-Gly-OMe in a dilute solution of methanol. Both studies have reasonable concurrence with respect to the preferred conformation of the hexapeptide and an analysis of the combined results suggests that the hexapeptide is stabilized by a beta-turn involving the Ala1,iC=O and Val4,iNH groups and a gamma-turn involving Gly5,iC=O and Gly3,iNH groups. A weaker interaction between Gly3,iC=O and Gly5,iNH groups is also found to be possible. Conformational features of the first valyl residue in the sequence Val-Ala-Pro-Gly-Val-Gly and the last valyl residue in Ala-Pro-Gly-Val-Gly-Val are compared and found to have similar torsion angles. The implications of such a similarity are discussed with respect to the conformation of the polyhexapeptide.

OBJECTIVE

To investigate the plasma amino acid spectrum in infants more than 1-year-old with neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) in order to identify potential diagnostic markers of NICCD.

METHODS

Infants less than 1 year of age who had been referred to our hospital for investigation of suspected conjugated hyperbilirubinemia between June 2003 and June 2009 were eligible for enrollment. A total of 182 infants were enrolled and divided into three groups: infants diagnosed with NICCD (n = 24), according to gene testing and/or western blotting results; infants diagnosed with biliary atresia (BA; n = 20), according to intra-operative cholangiography findings; and infants diagnosed with idiopathic neonatal intrahepatic hepatitis (INH; n = 138), according to exclusionary findings for diseases affecting the extrahepatic biliary system and no positive serology results to indicate infections with hepatitis B, C, A or E, toxoplasmosis, rubella, herpes simplex, human immunodeficiency virus-1, or syphilis. The plasma amino acid spectrum of each infant was analyzed by tandem mass spectrometry (MS/MS). The concentrations of 18 amino acids, as well as the ratio of each amino acid to total amino acids, were compared among the three groups. Selected ratios of amino acids were analyzed by receiver operating characteristic (ROC) curve analysis.

RESULTS

Compared with the BA and INH groups, the NICCD group had significantly lower levels of alanine (Ala; 175.7 and 205.7 vs. 136.3 mumol/L, P = 0.0001), aspartic acid (Asp; 47.5 and 43.1 vs. 31.55 mumol/L, P = 0.0041), glutamic acid (Glu; 276.16 and 263.24 vs. 175.71 mumol/L, P = 0.0075) and tryptophan (Trp; 41.90 and 47.97 vs. 28.51 mumol/L, P = 0.0003), but significantly higher levels of methionine (Met; 28.24 and 29.35 vs. 71.40 mumol/L, P = 0.0390), tyrosine (Tyr; 55.8 and 57.02 vs. 116.81 mumol/L, P = 0.0072) and citrulline (Cit; 15.09 and 15.65 vs. 97.42 mumol/L, P = 0.0001). The ratio of each amino acid to total amino acids showed the same trends for the NICCD group. The calculated areas under the ROC curves of the ratios of Cit, Tyr, and Met to the total amino acids were 0.874 (95% CI: 0.752 - 0.996), 0.814 (95% CI: 0.706 - 0.923), and 0.705 (95% CI: 0.535 - 0.875) respectively. The calculated area under the ROC curve of the ratio of Cit to Ala was 0.893 (95% CI: 0.781 - 1.005), and when the cut-off value of the ratio of Cit to Ala was 0.14 for diagnosis of NICCD, the sensitivity and specificity were 75% and 95% respectively.

CONCLUSIONS

The plasma amino acid spectrum may represent a diagnostic indicator for NICCD, particularly the ratio of Cit to Ala.

A 52-year male with past history of ulcerative colitis 20 years back (now in remission), developed recurrent small intestinal obstruction at intervals of a few months. CT scan did not detect the cause initially. A repeat CT scan (USA) showed interbowel fluid with transient ascites (serum albumin normal). Angio-oedema was suspected and low C4 with C1-esterase inhibitor (C1-INH) deficiency confirmed the diagnosis. Further investigation showed he was suffering from a chronic low grade small B cell lymphoma. He was treated with Rituximab 375 mg/m2 at intervals of one week for 4 weeks. He is asymptomatic with Transexamic acid (500 mg TDS) for last 1½ years.

Eluates of 13 malignant and 17 normal tissues were prepared at 56 degrees C using the continuous flow technique. Albumin was detected in all the eluates. IgG, IgA, C3 or haptoglobin were detected in most of the malignant and some of the normal tissues. Carcinoembryonic antigen, beta 2-microglobulin, alpha 1-antitrypsin or alpha 1-antichymotrypsin were detected in some of the eluates of the malignant tissues only. IgM, IgD, C1q, C4, Cl-INH, alpha 1-macroglobulin, beta 2-lipoprotein, fibrinogen and alpha 1-foetoprotein were not detected in any of the eluates. The ratio of the concentration of albumin to the concentration of IgG was similar in extracts and eluates of all the normal tissue and in 3 of the malignant tissues indicating non-specific binding of IgG.

OBJECTIVE

To explore effects of p, p'DDE on the expression of androgen binding protein (ABP), transferrin (Tf) and inhibin B (INH B) mRNA in testis Sertoli cells of Sprague Dawley rats.

METHODS

A method has been set up to obtain a large number of viable Sertoli cells from SD rats of 18-20 days of age. With a series of concentration p,p'-DDE (10, 30, and 50 micromol/L) co-incubating the Sertoli cells in vitro, the expression of ABP, Tf and INH B mRNA were determined by RT-PCR.

RESULTS

a) With increase of the incubated p, p'-DDE, the expression of ABP mRNA in Sertoli cells went up while that of Tf and INH B dropped in a dose-dependent manner (P < 0. 05). b) The correlation analysis among ABP, Tf and INH B showed that negative relationships were found between ABP and Tf or INH B, respectively (r = - 0. 391 3, P = 0. 032 5; r = - 0.235 2, P = 0.0158), and that positive correlation was indicated between Tf and INH B (r =0.4516, P =0.0047).

CONCLUSIONS

p,p'-DDE is a reproductive toxicant which disrupts the transcription of ABP, Tf and INH B in rat Sertoli cells so as to result in reproductive dysfunction.

The kinetic and thermodynamic stabilities of the group 13 hydrides EH(3) (E = B, Al, Ga, In, Tl, E113) are investigated by relativistic density functional and wave function based theories. The unimolecular decomposition of EH(3) ->> EH + H(2) becomes energetically more favorable going down the Group 13 elements, with the H(2)-abstraction of InH(3), TlH(3), and (E113)H(3) (E113: element with nuclear charge 113) being exothermic. In accordance with the Hammond-Leffler postulate, the activation barrier for the dissociation process decreases accordingly going down the group 13 elements in the periodic table shifting to an early transition state, with activation energies ranging from 88.4 kcal/mol for BH(3) to 41.3 kcal/mol for TlH(3) and only 21.6 kcal/mol for (E113)H(3) at the scalar relativistic coupled cluster level of theory. For both TlH(3) and (E113)H(3) we investigated spin-orbit effects using Dirac-Hartree-Fock and second-order Møller-Plesset theory to account for electron correlation. For (E113)H, spin-orbit coupling results in a chemically inert closed 7p(1/2)-shell, thus reducing the stability of the higher oxidation state even further. We also investigated the known organothallium compound Tl(CH(3))(3), which is thermodynamically unstable similar to TlH(3), but kinetically very stable with an activation barrier of 57.1 kcal/mol.

Although gamma/delta T-cells are known to contain the highest frequency of mycobacteria-reactive cells in humans, and recent studies have suggested that they play an important role in the initial immune response to Mycobacterium tuberculosis (Mtb), very few studies have attempted to analyze these cells in patients with active pulmonary tuberculosis (TB). The aim of the present study was therefore to evaluate the gamma/delta T-cell populations present in the peripheral blood and the IFN-gamma production of gamma/delta T-cells stimulated by PMA before and after anti-TB chemotherapy in patients in the initial treatment stage for primary active pulmonary TB. Cell populations were measured by three-color flow cytometry of peripheral blood mononuclear cells. We compared the population of gamma/delta T-cells and the production of IFN-gamma between normal healthy controls and TB patients. Absolute numbers of gamma/delta T-cells remained constant in the peripheral blood of TB patients. However, production of IFN-gamma in gamma/delta T-cells was dramatically suppressed prior to anti-TB chemotherapy in comparison with healthy control subjects, and further reduced following anti-TB chemotherapy. We also examined the influence of isoniazid (INH) in anti-TB chemotherapy. INH suppressed IFN-gamma production of gamma/delta and alpha/beta T-cells. The findings demonstrated a strong correlation between the production of IFN-gamma in gamma/delta T-cells and manifestation of primary active pulmonary TB, which was consistent with the hypothesized role for gamma/delta T-cells in the protective immune response to Mtb infection.

In order to know the adequate duration of the chemotherapy with two drugs (INH + RFP) for pulmonary tuberculosis with non-cavitary minimal radiological findings (minimal case), 278 cases with minimal lesion which had completed 9 months' chemotherapy, were observed for more than six months up to 5 years (mean duration = 54.4 months). Of them, 60 cases were bacteriologically confirmed by smear and/or culture examination. Many cases showed further improvement in radiological findings even after the end of the chemotherapy. Of 180 cases of initially infiltrative type (GAKKEN B type), 10 cases showed the enlargement of shadow radiologically, but were not regarded as relapsed cases, because they remained bacteriologically negative and the shadow improved in 1-2 months without additional chemotherapy. Only 3 cases (1.1%) were regarded as relapsed cases because of the positive bacteriological conversion and aggravation of the shadow. They were initially sputum negative. It can be concluded that for radiological minimal cases, nine months is enough for the duration of chemotherapy when the INH-RFP regimen is used.

A comparison is made of the data drawn from Hospital statistics for the period 1965-79 with regard to antibiograms from patients with first T. B. observations. A progressive reduction in primary resistance in the general sense was noted, with an increase in the case of RAMP only. There was a constant decrease for INH, PAS and ETB, whereas SM, KM and CS values were stationary. The conclusion is drawn that the reduction in primary resistance notwithstanding the continual employment of these drugs must be attributed to the fact that the resistant mycobacteria have depressed proliferative capacity and virulence. Primary resistance, therefore, while a good pointer to the epidemiology of T.B. infection, is not a cause for concern from the therapeutic standpoint.

In vitro culture has been used to study different aspects of ovarian function; however, this technique has not been applied to study recrudescence, or the return of ovarian function in seasonally breeding species. In Siberian hamsters, exposure to inhibitory photoperiods induces declines in ovarian function, which are restored with photostimulation. Because these changes are mediated by changes in systemic gonadotropin (GT) secretion, we hypothesized that culturing photoregressed ovaries with GT would restore aspects of function and induce expression of key folliculogenic factors. Adult female Siberian hamsters were exposed to either long-day (LD; 16L:8D) or short-day (SD; 8L:16D) photoperiods for 14 weeks to maintain in vivo cyclicity or induce gonadal regression, respectively. Isolated ovaries were then cultured for 10 days with or without GT. Ovarian mass and messenger RNA (mRNA) expression of mitotic marker Pcna were increased in cultured SD ovaries (cSD) ovaries with GT as compared to without GT, with no changes noted among cultured LD (cLD) ovaries. Media estradiol and progesterone concentrations increased in both cLD and cSD ovaries cultured with GT as compared to without GT. No differences in follicle numbers or incidence of apoptosis were noted across groups. In addition, differential mRNA expression of folliculogenic growth factors ( Bmp-4, Ntf-3, Inh-α, Gdf-9, Igf-1, Has-2, and Cox-2) was observed in cSD treated with or without GT. Together, these results suggest that this in vitro model could be a useful tool to (a) study the return of function in photoregressed ovaries, and (b) to identify the specific roles folliculogenic factors play in ovarian recrudescence.

BACKGROUND

Inhibins are dimeric glycoproteins, composed of an alpha-subunit (INH-α) and one of two possible beta-subunits (βA or βB), with substantial roles in human reproduction and in endocrine-responsive tumours. Aims of this study were to determine the serological measurement of inhibin A (α-βA) in breast cancer patients during chemotherapy.

METHODS

A series of 30 breast cancer patients who underwent standardised chemotherapy were prospectively evaluated before chemotherapeutic treatment as well as four weeks after chemotherapy and two years after chemotherapy for the serological expression of inhibin A. For statistical analysis the Wilcoxon rank sum test was used for paired samples. Statistical significance was assumed at p<0.05.

RESULTS

The concentration of inhibin A showed a significant decrease between data obtained before chemotherapy and after chemotherapy (p<0.005) and two-year follow-up (p<0.001). Interestingly, there were no differences in inhibin A concentrations between the four-week and two-year follow-up (p=0.744).

CONCLUSIONS

Chemotherapy significantly decreases inhibin A concentration during chemotherapy. This might reflect a suppression of ovarian function, being also a marker for chemotherapy-induced amenorrhoea. Moreover, it has been suggested that inhibin A might be a tumour marker for breast cancer, and therefore a sudden increase in its concentration might be indicative of breast cancer recurrence.

(<em>b</em>)Introduction</<em>b</em>): Hereditary angioedema due to C1 inhi<em>b</em>itor deficiency (C1-<em>INH</em>-HAE) is a rare yet still pro<em>b</em>a<em>b</em>ly underdiagnosed clinical condition. Recurrent episodes of su<em>b</em>cutaneous and su<em>b</em>-mucosal swelling may involve the skin, the gastrointestinal tract or even the upper airways, exposing the patients to the risk of death. With the aim of improving patients' quality of life, the therapeutic scenario has expanded over the years. (<em>b</em>)Areas covered</<em>b</em>): The focus of the present review is lanadeluma<em>b</em>, a fully human, κ-light-chain, monoclonal immunoglo<em>b</em>ulin G1 against plasma kallikrein, currently approved for long-term prophylaxis of C1-<em>INH</em>-HAE attacks in the USA and Canada and designated as an orphan drug <em>b</em>y the European Medicines Agency. (<em>b</em>)Expert opinion</<em>b</em>): Lanadeluma<em>b</em> is a<em>b</em>le to inhi<em>b</em>it plasma kallikrein with high selectivity and affinity. The su<em>b</em>sequent phases of drug development and the ongoing open-la<em>b</em>el trial have proven its safety and efficacy. It overcomes some of the limitations of other drugs availa<em>b</em>le for long-term prophylaxis, given the easy route of administration, the simple administration schedule and the possi<em>b</em>ility to tailor the treatment to each patient. Further studies are needed to test its efficacy also in other types of angioedema for which a central role of plasma kallikrein is envisaged.

Nonsteroidal anti-inflammatory medications (NSAIDs) are one of the most commonly used analgesics in the world. NSAIDs decrease prostaglandin synthesis through cyclooxygenase inhibition (COX-1 or COX-2). The effects of NSAIDs on survival and outcomes from global ischemia reperfusion events and specifically from cardiac arrest (CA) remain controversial. We hypothesized that NSAIDs prior to global whole-body ischemia reperfusion (I/R) injury impairs survival and outcomes. We explored this hypothesis in our swine model of Cardiac Arrest (CA) which involves global I/R with pretreatment using a predominantly COX-1 inhibitor (Indomethacin [COX-1/min COX-2 Inh], a COX-2 Inhibitor [COX-2-Inh, (Celecoxib)] or placebo control. We determined the effects of each inhibitor on a) survival, b) myocardial injury biomarker (Troponin 1), and c) Autonomic Nervous System (ANS) injury marker (heart rate variability [HRV]) up to 3 h after resuscitation. There were no survivals in COX-1/min COX-2-Inh pretreated animals and, 87% survived in both COX-2 Inhibited and control animals. COX-2 Inh pretreated animals had an 1800 fold increase of Troponin 1 compared to baseline whereas control animals had a 90 fold increase (p < 0.001). These results along with literature review of focal I/R in animal models with COX-2 overexpression, human studies of CA, and post myocardial infarction treatment with NSAIDs, support the hypothesis that NSAIDs prior to an I/R event impairs survival and outcomes. Specifically, predominantly COX-1 inhibition impairs survival, and COX-2 inhibition induces myocardial damage, autonomic nervous system dysfunction, and increases the risk for all-cause mortality and morbidity in humans post-MI which has significant implications for the nearly 10% of the population who are taking NSAIDs.

Two sets of diphenyl ether derivatives incorporating five-mem<em>b</em>ered 1,3,4-oxadiazoles, and their open-chain aryl hydrazone analogs were synthesized in good yields. Most of the synthesized compounds showed promising in vitro antimyco<em>b</em>acterial activity against <i>Myco<em>b</em>acterium tu<em>b</em>erculosis</i> H37Rv. Three diphenyl ether derivatives, namely hydrazide (<em>b</em>)3</<em>b</em>), oxadiazole (<em>b</em>)4</<em>b</em>) and naphthylarylidene (<em>b</em>)8g</<em>b</em>) exhi<em>b</em>ited pronounced activity with minimum inhi<em>b</em>itory concentrations (MICs) of 0.61, 0.86 and 0.99 μg/mL, respectively compared to triclosan (10 μg/mL) and isoniazid (<em>INH</em>) (0.2 μg/mL). Compounds (<em>b</em>)3</<em>b</em>), (<em>b</em>)4</<em>b</em>), and (<em>b</em>)8g</<em>b</em>) showed the InhA reductase enzyme inhi<em>b</em>ition with higher IC<su<em>b</em>)50</su<em>b</em>) values (3.28-4.23 µM) in comparison to triclosan (1.10 µM). Correlation <em>b</em>etween calculated physicochemical parameters and <em>b</em>iological activity has <em>b</em>een discussed which justifies a strong correlation with respect to the inhi<em>b</em>ition of InhA reductase enzyme. Molecular modeling and drug-likeness studies showed good agreement with the o<em>b</em>tained <em>b</em>iological evaluation. The structural and experimental information concerning these three InhA inhi<em>b</em>itors will likely contri<em>b</em>ute to the lead optimization of new anti<em>b</em>iotics for <i>M. tu<em>b</em>erculosis</i>.

Keywords: H37Rv; InhA inhibitors; antitubercular agents; diphenyl ether; molecular docking.

Both isoniazid (INH) and cefazolin (CEZ) can have serious adverse effects on the central nervous system (CNS), causing seizures. In this study, we investigated the effect of INH on the pharmacodynamics of CEZ-induced seizures in rats. Male Wistar rats pretreated with INH (150 mg/kg i.p.) or saline received an intravenous infusion of CEZ at 3.2 g/h/rat until the onset of seizures, then samples of cerebrospinal fluid (CSF), blood (for serum), and brain were obtained immediately. The administration of INH was associated with a reduction in the total dose of CEZ required to produce seizures. The concentrations of CEZ in serum, brain, and CSF in INH-treated rats at the onset of seizures were significantly lower than those in control rats. In rats coadministered with pyridoxine (150 mg/kg s.c.), the concentration of CEZ in CSF at the onset of seizures was significantly higher than that in rats administered INH only. These results suggest that INH potentiates the sensitivity of the CNS to CEZ-induced seizures, and that the increased sensitivity is associated with the inhibition of vitamin B(6) metabolism by INH.

The transforming growth factors <em>beta</em> (TGFβ) are local factors produced by ovarian cells which, after binding to their receptors, regulate follicular deviation and ovulation. However, their regulation and function during corpus luteum (CL) regression has been poorly investigated. The present study evaluated the mRNA regulation of some TGFβ family ligands and their receptors in the bovine CL during induced luteolysis <i>in vivo</i>. On day 10 of the estrous cycle, cows received an injection of prostaglandin F2α (PGF) and luteal samples were obtained from separate groups of cows (n= 4-5 cows per time-point) at 0, 2, 12, 24 or 48 h after treatment. Since TGF <em>beta</em> family comprises more than 30 ligands, we focused in some candidates genes such as activin receptors (<i>ACVR-1A</i>, <i>-1B</i>, <i>-2A</i>, <i>-2B</i>) <i>AMH</i>, <i>AMHR2</i>, <i>BMPs</i> (<i>BMP-1</i>, <i>-2</i>, <i>-3</i>, <i>-4</i>, <i>-6 and -7</i>), BMP receptors (<i>BMPR-1A</i>, <i>-1B</i> and -<i>2</i>), inhibin subunits (<i><em>INH</em>-A</i>, <i>-BA</i>, <i>-BB</i>) and <em>beta</em>glycan (<i>TGFBR3</i>). The mRNA levels of <i>BMP4</i>, <i>BMP6</i> and <i><em>INH</em>BA</i> were higher at 2 h after PGF administration (P<0.05) in comparison to 0 h. The relative mRNA abundance of <i>BMP1</i>, <i>BMP2</i>, <i>BMP3</i>, <i>BMP4</i>, <i>BMP6</i>, <i>ACVR1B</i>, <i><em>INH</em>BA</i> and <i><em>INH</em>BB</i> was upregulated up to 12 h post PGF (P<0.05). On the other hand, <i>TGFBR3</i> mRNA that codes for a reservoir of ligands that bind to TGF-<em>beta</em> receptors, was lower at 48 h. In conclusion, findings from this study demonstrated that genes encoding several TGFβ family members are expressed in a time-specific manner after PGF administration.

The mechanisms by which GnIH regulates the steroid synthesis pathway in duck granulosa cells remain poorly understood. In this study, we measured steroid hormone secretion by ELISA and reproduction-associated gene expression by quantitative real-time Polymerase Chain Reaction (qPCR) in duck granulosa cells treated with different concentrations of GnIH (0, 0.1, 1, 10, and 100 ng/mL) for 24 h. The genome-wide expression profiles of GnIH-treated cells (0 and 10 ng/mL) were evaluated by high-throughput RNA sequencing. Compared with untreated cells, the secretion of the steroid hormones E2, E1, P4, and T was downregulated, with that of E1 and P4 reaching statistical significance (<i>P</i><0.05); in contrast, the secretion of ACV and <em>INH</em> was significantly upregulated (<i>P</i><0.05) after treatment with 10 and 100 ng/mL GnIH. The expression of encoding steroidogenic proteins and enzymes genes (<i>STAR</i>, <i>CYP11A1</i>, <i>CYP17A1</i>, <i>CYP19A1</i>, and <i>3-<em>β</em>-HSD</i>) and encoding gonadotropin receptors genes (<i>FSHR</i>, <i>LHR</i>) were significantly declined (<i>P</i><0.05) in the 10 and 100 ng/mL GnIH treatments. Transcriptome sequencing identified 348 differentially expressed genes (DEGs), including 253 upregulated and 95 downregulated genes. The DEGs were mainly involved in cell growth and death, immune response, and steroid biosynthesis pathways. We identified four novel DEGs (<i>MROH5</i>, <i>LOC113840576</i>, <i>SDR42E1</i>, and <i>LOC113841457</i>) with key roles in the regulation of steroid hormone biosynthesis. Our study revealed changes in gonadal steroid hormone secretion and steroid biosynthesis pathway-related gene expression in duck granulosa cells under the inhibitory effect of GnIH. These data contribute to our understanding of the molecular and genetic mechanisms underlying reproduction in ducks.

Keywords: Gonadotropin-inhibitory hormone (GnIH); gene expression; granulosa cell; reproduction; steroid synthesis.

The incidence rate of tuberculosis (TB) in patients with human immunodeficiency virus (HIV) infection is 26 times higher than that in other patients. Patients with both infections require long-term combination therapy, which increases therapy complexity and might lead to serious adverse reactions and drug-drug interactions. To optimize therapy for patients with HIV and TB coinfection, we developed an ultra-high-performance liquid chromatography/tandem mass spectrometry (UHPLC-MS/MS) method to simultaneously quantify four anti-tuberculosis drugs and one isoniazid (INH) metabolite. Blood samples (n = 32) from 16 patients with HIV and TB coinfection were collected. Plasma protein precipitation with acetonitrile was followed by a hydrazine reaction between INH and cinnamaldehyde (CA) to produce phenylhydrazone (CA-INH) and dilution with heptafluorobutyric acid. The separation was performed on an Acquity UHPLC HSS T3 1.8 μm column (2.1 × 100 mm, Waters) with a mobile phase consisting of 10 mmol/L ammonium formate (pH = 4) in water (solvent A) and 0.1 % formic acid in methanol (solvent B) in a gradient elution. The compounds were detected using a positive multiple reaction monitoring model. INH, acetyl-INH (AC-INH), rifampicin (RIF), ethambutol (EMB), and pyrazinamide (PZA) showed good linear relationships in their quantitative ranges, with lower limits of quantification of 48, 192, 200, 96, and 480 ng/mL, respectively. The inter- and intraday precision was within 15 %, and the accuracy was between 85 % and 115 %. The mean plasma concentrations of INH, AC-INH, RIF, EMB, and PZA in patients were 1990.23 (24-16 600), 863.06 (96-2880), 3507.05 (229-9800), 808.10 (149-2130), and 18 838.33 (240-34 800) ng/mL, respectively. The plasma concentrations detected in the 16 patients were lower than the targeted concentrations in HIV-negative TB patients. In summary, we developed a simple UHPLC-MS/MS method for simultaneous quantification of first-line TB drugs, and successfully applied it for therapeutic drug monitoring in patients with HIV and TB coinfection. This method will facilitate monitoring of TB drugs in the future.

Keywords: Ethambutol; First-line anti-tuberculosis drugs; Human immunodeficiency virus-tuberculosis; Isoniazid; Rifampicin; Ultra-high-performance liquid chromatography/tandem mass spectrometry.

Hypertension is the leading risk factor for the development of heart diseases and stroke. Many hypertensive patients experience undesira<em>b</em>le side effects to conventional antihypertensive pharmacotherapy. Cil et al. documented the antihypertensive profile of a novel molecule, TM<su<em>b</em>)<em>inh</em></su<em>b</em>)-23 (2-<em>b</em>romodifluoroacetylamino-5,6,7,8-tetrahydro-4H-cyclohepta[<em>b</em>]thiophene-3-car<em>b</em>oxylic acid o-tolylamide), in the spontaneously hypertensive rat model of systemic hypertension. They showed that this agent reduces <em>b</em>lood pressure <em>b</em>y <em>inh</em>i<em>b</em>iting transmem<em>b</em>rane mem<em>b</em>er 16A-encoded calcium-activated chloride channels in vascular myocytes from resistance arteries. If validated, TM<su<em>b</em>)<em>inh</em></su<em>b</em>)-23 could <em>b</em>ecome a useful clinical tool.