(<em>b</em>)O<em>b</em>jective</<em>b</em>) The aim of this study was to evaluate rates of preterm <em>b</em>irth (PTB) and o<em>b</em>stetric complication with maternal serum analytes > 2.5 multiples of the median (MoM) <em>b</em>y degree of elevation. (<em>b</em>)Study Design</<em>b</em>) Retrospective cohort study of singleton live-<em>b</em>irths participating in the California Prenatal Screening Program (2005-2011) examining PTB and o<em>b</em>stetric complication for α-fetoprotein (AFP), human chorionic gonadotropin (hCG), unconjugated estriol (uE3), and inhi<em>b</em>in A (<em>INH</em>) <em>b</em>y analyte su<em>b</em>group (2.5 to < 6.0, 6.0 to < 10.0, and ≥ 10.0 MoM vs. < 2.5 MoM). (<em>b</em>)Results</<em>b</em>) The risk of o<em>b</em>stetric complication increased with increasing hCG, AFP, and <em>INH</em> MoM, and were greatest for AFP and <em>INH</em> of 6.0 to <10.0 MoM. The greatest risk of any adverse outcome was seen for hCG MoM ≥ 10.0, with relative risk (RR) of PTB < 34 weeks of 40.8 (95% confidence interval [CI]: 21.7-77.0) and 13.8 (95% CI: 8.2-23.1) for o<em>b</em>stetric complication. (<em>b</em>)Conclusions</<em>b</em>) In euploid, structurally normal fetuses, all analyte elevations > 2.5 MoM confer an increased risk of PTB and, except for uE3, o<em>b</em>stetric complication, and risks for each are not uniformly linear. These data can help guide patient counseling and antenatal management.

Keywords: AFP; adverse pregnancy outcomes; estriol; hCG; inhibin.

Isoniazid (<em>INH</em>), a first-line drug in anti-tu<em>b</em>erculosis therapy, is known to <em>b</em>e potentially harmful and is associated with numerous side effects especially in the <em>b</em>lood and liver. In the course of our previous investigations, 1,2,3-thiadiazole containing hydrazone (compound (<em>b</em>)3</<em>b</em>)) showed excellent antimyco<em>b</em>acterial activity against a referent strain <i>M. tu<em>b</em>erculosis</i> H37Rv (MIC value 0.39 μM), low cytotoxicity, and did not have toxic effects when administered <em>b</em>y oral or intraperitoneal routes to experimental animals (selectivity index SI > 1979, LD<su<em>b</em>)50</su<em>b</em>)>2000 mg/kg <em>b</em>.w.) what revealed its suita<em>b</em>ility for further exploration. In the present study compound (<em>b</em>)3</<em>b</em>) was chosen to determine its effects on the liver and kidney functions in female mice. The compound was administered orally for 14 days at three doses (100, 200, and 400 mg/kg <em>b</em>.w.). The quantity of malondialdehyde (MDA), the level of reduced glutathione (GSH), <em>b</em>lood hematological and <em>b</em>iochemical parameters were assessed, and urine analysis was carried out. As a positive control <em>INH</em> was used orally at a dose of 50 mg/kg <em>b</em>.w. The investigated compound (<em>b</em>)3</<em>b</em>) did not affect the urine and serum hematological and <em>b</em>iochemical parameters as <em>INH</em> did, compared to those of the control mice. The new compound did not affect significantly the MDA quantity and maintained its level near to the control values, though lower <em>b</em>y 36% (<i>p</i> < 0.05) than in the <em>INH</em> treated animals. At the higher doses, 200 and 400 mg/kg, it depleted the GSH content <em>b</em>y 25% (<i>p</i> < 0.05), compared to the control. However, its level remained 47% (<i>p</i> < 0.05) higher than in the <em>INH</em> treated animals.

The explosive global spreading of multidrug resistant Mycobacterium tuberculosis (Mtb) has provoked an urgent need to discover novel anti-TB agents. Enoyl-acyl carrier protein reductase from Mtb is a well-known and thoroughly studied target for anti-tuberculosis therapy. In the present analysis, virtual screening techniques performed from Drug bank database by utilizing INH-NAD adduct as query for the discovery of potent anti-TB agents. About 100 molecules sharing similar scaffold with INH-NAD adduct were analyzed for their binding effectiveness. The initial screening based on number of rotatable bonds gave 42 hit molecules. Subsequently, physiochemical properties such as toxicity, solubility, drug-likeness and drug score were analyzed for the filtered set of compounds. Final data reduction was performed by means of molecular docking and normal mode docking analysis. The result indicates that DB04362, adenosine diphosphate 5-(beta-ethyl)-4-methyl-thiazole-2-carboxylic acid could be a promising lead compound and be effective in treating sensitive as well as drug-resistant strains of Mtb. We believe that this novel scaffolds might be the good starting point for lead compounds and certainly aid the experimental designing of anti-tuberculosis drug in a short time.

Isonicotinic acid hydrazide (Isoniazid, INH) is one of the most commonly used first-line anti-tuberculosis drugs, which has been reported that the high concentration of INH in human body can lead to epilepsy, liver function failure, and even death. Therefore, studying the potential binding effects of INH on the structure and activity of human serum albumin (HSA) and catalase (CAT) is very essential for evaluating its toxicity and side effect. In this paper, multi-spectroscopic and molecular docking methods were used to elucidate the patterns of INH to HSA and CAT under imitated physiological conditions. The inner filter effect of all fluorescence data in the paper was eliminated to get accurate binding parameters. The Stern-Volmer quenching constants (KSV) of both HSA-INH system and CAT-INH system inversely correlated with temperatures, demonstrating that INH quench the intrinsic fluorescence of HSA and CAT via static quenching. The conformational investigation of HSA and CAT through UV-visible absorption spectroscopy, synchronous fluorescence and circular dichroism (CD) showed that INH could change the micro-environment of tryptophan residues and reduced the α-helix content of protein. These results demonstrated that the binding of INH may lead to the loosening of protein skeleton, which which may affect its physiological function. The results of molecular docking revealed that the INH was located in Sudlow’s site I of HSA. And INH bound to CAT at a cavity among the wrapping domain helical domain and β-barrel, which resulted in the inhibition of CAT activity. In addition, Levofloxacin (LVFX) is a new effective and safe second-line anti-tuberculosis drugs and can improve the curative effect on anti-TB by using with other anti-TB drugs, the result of Hill’s coefficients (nH) about synergy between INH and proved that LVFX promoted the interaction of HSA with INH. Moreover, according to the CD spectra, synergy between INH and LVFX changed the conformation of HSA and the amount of α-helix decreased about 7.9%. This work will provide important insights into the binding and toxicity mechanism of INH to HSA and CAT in vivo and is expected to be helpful in evaluating the essential information for using the INH safely.

<em>β</em>-nicotinamide adenine dinucleotide (NAD⁺) is a critical molecule that is involved in multiple cellular functions. The cluster of differentiation (CD) 38 is a multifunctional enzyme with NAD⁺ nucleosidase activity. Our previous work revealed the CD38-dependent interactions of isoniazid (<em>INH</em>), an anti-tu<em>b</em>erculosis drug, with NAD⁺ to form <em>INH</em>-NAD adduct. In the current work, our meta<em>b</em>olomic analysis discovered a novel NAD⁺ adduct with acetylisoniazid (Ac<em>INH</em>), a primary <em>INH</em> meta<em>b</em>olite mediated <em>b</em>y <i>N</i>-acetyltransferase (NAT), and we named it Ac<em>INH</em>-NAD. Using Nat1/2(-/-) and Cd38(-/-) mice, we determined that Ac<em>INH</em>-NAD formation is dependent on <em>b</em>oth NAT and CD38. Because NAT is expressed in hepatocytes (HP), whereas CD38 is expressed in Kupffer cells (KC) and hepatic stellate cells (HSC), we explored cell type-specific roles of CD38 in the formation of Ac<em>INH</em>-NAD as well as <em>INH</em>-NAD. We found that <em>b</em>oth <em>INH</em>-NAD and Ac<em>INH</em>-NAD were produced in the incu<em>b</em>ation of <em>INH</em> or Ac<em>INH</em> with KC and HSC, <em>b</em>ut not in HP. These data suggest that hepatic non-parenchymal cells, such as KC and HSC, are the major cell types responsi<em>b</em>le for the CD38-dependent interactions of <em>INH</em> with NAD⁺ in the liver. (<em>b</em>)Significance Statement</<em>b</em>) The current study identified Ac<em>INH</em>-NAD as a novel meta<em>b</em>olite of <em>INH</em> in the liver. Our work also revealed the essential roles of non-parenchymal cells including KC and HSC in the CD38-dependent interactions of NAD⁺ with <em>INH</em>, leading to the formation of <em>b</em>oth <em>INH</em>-NAD and Ac<em>INH</em>-NAD in the liver. These data can <em>b</em>e used to guide the future studies on the mechanisms of <em>INH</em> and NAD⁺ interactions and their contri<em>b</em>utions to <em>INH</em>-induced liver injury.

Keywords: drug metabolism; hepatocytes; kupffer cells; metabolomics.

According to the recent advances of molecular biological technics, some of the genetic mechanisms of drug-resistance of Mycobacteria has been uncovered. Generally, drug-resistance of Mycobacterium tuberculosis was caused by point mutations in chromosomal gene. In isoniazid (INH) resistant M. tuberculosis, mutations and genetic deletions in catalase-peroxidase gene (katG), inhA gene, or alkyl hydroperoxide reductase gene were reported. On the other hand, mutations and other genetic alterations in RNA polymerase beta subunit gene (rpoB) were the major mechanisms of resistance to rifampicin (RFP) with high frequencies of 90% or more. Moreover, these genetic alterations in rpoB gene were suspected as the resistant mechanism to other rifamycin antituberculosis drugs, such as rifabutin. In addition, it was reported that point mutations in 16S rRNA gene (rrs) and ribosomal protein S12 gene (rpsL) induced M. tuberculosis as streptomycin (SM) resistant phenotype. Furthermore, nicotinamidase (pncA) gene, DNA gyrase A subunit (gyrA) gene, and embB gene were reported as the responsible gene to pyrazinamide-, quinolone- and ethambutol-resistance, respectively. Although all mechanisms of drug-resistance were still unclear, these information are very useful and helpful for development of rapid diagnosis system of drug-resistant M. tuberculosis.

Isoniazid (INH) is a first-line antituberculosis drug. The incidence of adverse reactions accompanied by inflammation in the liver during drug administration to tuberculosis patients is high and severely affects clinical treatment. To better understand the mechanism of hepatotoxicity induced by INH under the inflammatory state, we compared the differences in levels of hepatotoxicity from INH between normal zebrafish and zebrafish in an inflammatory state to elucidate the hepatotoxic mechanism using different endpoints such as mortality, malformation, inflammatory effects, liver morphology, histological changes, transaminase analysis, and expression levels of certain genes. The results showed that the toxic effect of INH in zebrafish in an inflammatory state was more obvious than that in normal zebrafish, that liver size was significantly decreased as measured by liver fatty acid binding protein (LFABP) reporter fluorescence and intensity, and that alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were significantly increased. Hematoxylin and eosin (HE) staining and electron microscopy showed that hepatocyte injury was more obvious in the inflammatory state. In the inflammatory state, INH significantly increased the expression levels of endoplasmic reticulum stress (ERS)-related factors (GRP78, ATF6, PERK, IRE1, XBP1s, GRP94, and CHOP), autophagy-related factors (beclin 1, LC3, Atg3, and Atg12), and apoptosis-related factors (caspase-3, caspase-8, caspase-9, Bax, p53, and Cyt) in larvae. Correlational analyses indicated that the transcription levels of the inflammatory factors interleukin-1b (IL-1b), tumor necrosis factor beta (TNF-β), cyclooxygenase 2 (COX-2), and TNF-ɑ were strongly positively correlated with ALT and AST. Furthermore, the ERS inhibitor sodium 4-phenylbutyrate (4-PBA) could ameliorate the hepatotoxicity of INH-lipopolysaccharide (LPS) in zebrafish larvae. These results indicated that INH hepatotoxicity was enhanced in the inflammatory state. ERS and its mediated autophagy and apoptosis pathways might be involved in INH-induced liver injury promoted by inflammation.

A series of pyrazolo[1,5-<i>a</i>]pyridine-3-car<em>b</em>oxamide (PPA) derivatives <em>b</em>earing diaryl side chain was designed and synthesized as new antitu<em>b</em>erculosis agents, aiming to improve the efficacy toward drug resistant <i>Myco<em>b</em>acterium tu<em>b</em>erculosis</i> (<i>Mt<em>b</em></i>) strains. Most of the su<em>b</em>stituted diphenyl and heterodiaryl PPAs exhi<em>b</em>ited excellent <i>in vitro</i> potency against the drug susceptive H37Rv strain (MIC < 0.002-0.381 μg/mL) and drug resistant <i>Mt<em>b</em></i> strains (<em>INH</em>-resistant (r<em>INH</em>), MIC < 0.002-0.465 μg/mL; RMP-resistant (rRMP), MIC < 0.002-0.004 μg/mL). Noticea<em>b</em>ly, some compounds also showed very low cytotoxicity against Vero cells. Further, compound (<em>b</em>)6j</<em>b</em>) displayed good pharmacokinetic profiles with oral <em>b</em>ioavaila<em>b</em>ility (F) of 41% and significantly reduced the <em>b</em>acterial <em>b</em>urden in an autoluminescent H37Ra infected mouse model.

BACKGROUND

Alternative treatment strategies have become essential in overcoming the problem of drug-resistant Mycobacterium tuberculosis (Mtb). In this preliminary in vitro study, the anti-tuberculosis (anti-TB) activity of exogenous iron chelators (xenosiderophores) such as Exochelin-MS (Exo-MS) and Deferoxamine-B (DFO-B) was evaluated against ten multi-drug-resistant (MDR) and seven pyrazinamide-resistant (PZA R ) Mtb isolates.

METHODS

Mycobacteria Growth Indicator Tube-Drug Susceptibility Test was used to assess the anti-TB effect of Exo-MS or DFO-B individually and their combinations with isoniazid (INH), rifampicin (RIF) and pyrazinamide (PZA).

RESULTS

For the MDR-Mtb isolates, Exo-MS alone inhibited two out of the five isolates tested. Whereas, DFO-B alone inhibited nine out of the ten MDR isolates tested. For PZA-resistant Mtb isolates, both Exo-MS and DFO-B individually inhibited five out of the seven isolates. The MIC of Exo-MS in combination with INH, RIF and PZA remained the same. The MIC of DFO-B decreased when tested in combination with INH, RIF and PZA.

CONCLUSIONS

Exo-MS and DFO-B were shown to have activity against drug-resistant Mtb isolates. Therefore, these xenosiderophores may be useful adjuncts to antibiotics in overcoming the problem of drug-resistant Mtb in clinical setting.

(<em>b</em>)INTRODUCTION:</<em>b</em>) Resistance to first-line anti-tu<em>b</em>erculosis drugs is a major concern in the treatment of the disease. New strategies, such as the use of efflux pump inhi<em>b</em>itors (EPIs), are <em>b</em>eing investigated to improve the outcome of the treatment. Verapamil (VP), one such inhi<em>b</em>itor, was shown to inhi<em>b</em>it several efflux pump (EP) <i>Myco<em>b</em>acterium tu<em>b</em>erculosis</i> proteins and demonstrate synergic activity with anti-TB drugs.(<em>b</em>)OBJECTIVE:</<em>b</em>) To evaluate the com<em>b</em>inatory effect of isoniazid (<em>INH</em>) and VP in <i>M. tu<em>b</em>erculosis</i>.(<em>b</em>)METHODS:</<em>b</em>) Minimal inhi<em>b</em>itory concentrations and com<em>b</em>inatory effects of <em>INH</em>+VP were determined using respectively resazurin microtitre assay plate (REMA) and resazurin drugs com<em>b</em>ination microtitre assay (REDCA). From the results, we selected three <em>b</em>acilli with different suscepti<em>b</em>ility profiles and assessed their expression of 10 EP genes through quantitative reverse transcription polymerase chain reaction after exposure to <em>INH</em>, VP and <em>INH</em> + VP for 48 h.(<em>b</em>)RESULTS:</<em>b</em>) A significant reduction of <em>INH</em> MIC was o<em>b</em>served in <em>INH</em>-suscepti<em>b</em>le isolates upon com<em>b</em>ination with VP. In <em>b</em>rief, gene expression assays revealed expression patterns that could <em>b</em>e correlated with each resistance profile, presence or a<em>b</em>sence of gene mutations and com<em>b</em>inatory effect with VP.(<em>b</em>)CONCLUSION:</<em>b</em>) Com<em>b</em>ining VP with <em>INH</em> showed important results in drug-suscepti<em>b</em>le strains, and clinical trials on com<em>b</em>ined VP + anti-TB drugs should <em>b</em>e discussed.

The thirty-two times of treatment in 27 patients with multidrug-resistant tuberculosis (MDR-TB) were analyzed retrospectively. In twenty-eight times of treatments cases had previous histories of antituberculosis chemotherapy. Drug sensitivity tests were performed by Microtiter method for isoniazid (INH), rifampicin (RFP), ethambutol, streptomycin, kanamycin, enviomycin, ethionamide, para-aminosalicylic acid and cycloserine. A drug is defined as 'active drug' when the drug was proved to be sensitive by the drug sensitivity tests or never used in the past or used for not more than 2 months in case of pyrazinamide (PZA) and less than one month for fluoroquinolones. Outcomes of treatments were grouped as follows; A: bacteriologically negative for more than 24 months, B: bacteriologically negative for more than 6 months but less than 24 months, C: bacteriologically relapsed after negative conversion, D: continuously bacilli positive for M. tuberculosis. Mean age of patients in each group were; 61.0 yrs for group A (n = 10), 61.0 yrs for group B (n = 7), 52.5 yrs for group C (n = 4), 57.9 yrs for group D (n = 11). All patients had cavitary disease and positive sputum smears for acid-fast bacilli. Mean numbers of 'active drugs' used per treatment in each group, were 3.6, 3.3, 2.5 and 1.8 respectively, while the mean number of resistant drug including INH and RFP were 2.8, 3.3, 2.5 and 3.7. The number of drugs, which was unable to use due to toxicity, were 0.20, 0.14, 0.50, and 0.73 per treatment respectively. All of 9 patients treated with four 'active drugs' were in group A or B and succeeded to achieve negative conversion. The duration of chemotherapy in group A was 13 to 44 months. Treatment had failed in 4 out of 11 patients treated with 3 'active drugs' and 11 out of 12 patients treated with less than 2 'active drugs'. Fluoroquinolones (ofloxacin, levofloxacin or sparfloxacin) were used in 7 out of 10 patients in group A and in 6 out of 9 patients treated with four-drug regimens while they were used only in 3 out of 11 patients in group D. Regimens with at least 4 sensitive drugs are mandatory for the successful treatment of MDR-TB and fluoroquinolones are needed in the majority of cases to ensure the four-drug regimen, because of frequent drug resistance or toxicity to other antituberculosis drugs.

Background: Lupus anticoagulant (LA) can be a cause of thrombosis and/or pregnancy morbidities, producing antiphospholipid syndrome (APS). An increase in thrombin generation (TG) is correlated with prothrombotic status. Several changes in TG-derived parameters have been reported in APS patients.

Objectives: Evaluate whether the TG phenotype of APS can also be described in LA subjects without clinical manifestations of APS, and to investigate the possible influence of both LA potency and antiphospholipid (aPL) profile on it.

Results: TG was analyzed in 153 cases of LA and 41 healthy controls. We have observed prolongation of both lag time (3.7 min vs 2.32 min, p < 0.001) and time to peak (6.48 min vs 5.27 min, p < 0.001), increased peak height (221.7 nM vs 182.7 nM, p < 0.001), slightly higher ETP (221.7 nM·min vs 182.7 nM·min, p = 0.041), and higher velocity index (100.7 nM/min vs 74.53 nM/min, p = 0.001) in LA subjects compared to controls. After adding thrombomodulin (TM), ETP%inh was significantly lower in LA group (37.90% vs 59.90%, p < 0.001) showing resistance to TM/activated protein C (APC). Significant differences were found in lag time, time to peak and ETP%inh according to the potency and aPL profile.

Conclusions: Previously described differences in TG-derived parameters in APS patients have been confirmed in incidental LA subjects: prolonged lag time and time to peak, slightly higher ETP, higher peak height, and less sensitivity to TM/APC. High LA potency and triple-positive aPL profile enhance differences in lag time, time to peak and, especially, increase APC resistance, but no effect in ETP was observed.

Keywords: Anti-β(2)GPI; Anticardiolipin; Antiphospholipid; Lupus anticoagulant; Thrombin generation.

OBJECTIVE

To study the clinical features of BCG infection in children.

METHODS

51 cases confirmed with BCG infection from all over China were enrolled and followed up for at least 6 months. All cases were treated with anti-tuberculosis drugs. A random, open, group control study was designed in non-disseminated cases to evaluate curative effects of anti-tuberculosis drugs for early stage BCG infection. Disseminated cases were also closely monitored, and patients were given combined anti-tuberculosis drug therapy.

RESULTS

In 34 (66.7%) non-disseminated cases, 19 children with local infections were treated with Isoniazid (Group A) and 15 were treated with Isoniazid and Rifampin (Group B). In the first 3 months, Group B responded better to anti- tuberculosis drug therapy than Group A (P<0.05). At the end of 6 months drug therapy, improvement rate was 100% of Group B vs. 89.5% of Group A (P<0.05). 33.3% children were admitted with disseminated BCG disease and were initially treated with Isoniazid and Rifampin. Most of these children responded poorly to drug therapies: Both isolated strains and BCG vaccination strain showed resistance to isoniazid, but susceptible to other First-line anti-tuberculosis drugs (Rifampin, Ethambutol and Streptomycin).

CONCLUSIONS

INH does not perform well for treating BCG Chinese infections. Multiple drug regimens are necessary for treatment and preventing Drug-Resistance. Even for non-disseminated cases, preventive therapy using mono-isoniazid regimen is not suitable. BCG infections also occur in children without clear immunodeficiency, so parental education and awareness of health-care workers is essential for promptly recognition and handling BCG infections.

Impaired regulatory T-cell (Treg) responses and upregulated interleukin-1 receptor-associated kinase 1 (IRAK1) expression are associated with the development of human systemic lupus erythematosus (SLE). Here, we show that the levels of upregulated IRAK1 expression in circulating Tregs are correlated with the percentages of apoptotic Tregs, Systemic Lupus Erythematosus Disease Activity Index scores, and serum complement C3 levels in SLE patients. High levels of plasma interferon (IFN)-α in SLE patients induced IRAK1 phosphorylation to trigger Treg apoptosis, which was mitigated by IRAK1 inhibitor (IRAK-Inh) treatment. Bioinformatics indicated that IRAK1 activation was related to the IFN-α/β and mitogen-activated protein kinase (MAPK) signaling in Tregs and IFN-α treatment induced the p38 and MAPK/ERK kinase 3/6 phosphorylation, which was attenuated by IRAK-Inh in Tregs. Treatment with IRAK-Inh effectively alleviated renal injury and promoted the survival of lupus-prone B6.MRL-Fas^lpr /Nju mice. Therefore, IFN-α induced IRAK1 activation to promote Treg apoptosis, contributing to the pathogenesis of SLE and IFN-α/IRAK1 may be therapeutic targets for SLE.

Keywords: apoptosis; interferon-α; interleukin-1 receptor-associated kinase 1; regulatory T cells; systemic lupus erythematosus.

Clostridia can cause hepatic damage in domestic livestock, and wild and laboratory animals. Clostridium novyi type B causes infectious necrotic hepatitis (INH) in sheep and less frequently in other species. Spores of C. novyi type B can be present in soil; after ingestion, they reach the liver via portal circulation where they persist in phagocytic cells. Following liver damage, frequently caused by migrating parasites, local anaerobic conditions allow germination of the clostridial spores and production of toxins. C. novyi type B alpha toxin causes necrotizing hepatitis and extensive edema, congestion, and hemorrhage in multiple organs. Clostridium haemolyticum causes bacillary hemoglobinuria (BH) in cattle, sheep, and rarely, horses. Beta toxin is the main virulence factor of C. haemolyticum, causing hepatic necrosis and hemolysis. Clostridium piliforme, the causal agent of Tyzzer disease (TD), is the only gram-negative and obligate intracellular pathogenic clostridia. TD occurs in multiple species, but it is more frequent in foals, lagomorphs, and laboratory animals. The mode of transmission is fecal-oral, with ingestion of spores from a fecal-contaminated environment. In affected animals, C. piliforme proliferates in the intestinal mucosa, resulting in necrosis, and then disseminates to the liver and other organs. Virulence factors for this microorganism have not been identified, to date. Given the peracute or acute nature of clostridial hepatitis in animals, treatment is rarely effective. However, INH and BH can be prevented, and should be controlled by vaccination and control of liver flukes. To date, no vaccine is available to prevent TD.

OBJECTIVE

To evaluate the predictive value of serum inhibin B (INH B) levels as an indicator of the presence of testicular spermatozoa in nonobstructive azoospermia.

METHODS

Forty patients with nonobstructive azoospermia (NOA), 20 patients with obstructive azoospermia (OA), and 10 fertile volunteers were involved in this study. A chemoluminescence method was used to measure the levels of FSH; Inhibin B was analysed by using sandwich enzyme-linked immuno-sorbent assay.

RESULTS

Patients with nonobstructive azoospermia has significantly higher levels of serum FSH [(21.34 +/- 12.15) IU/L] and significantly lower levels of inhibin B [(53.15 +/- 58.74) ng/L] than patients with obstructive azoospermia [FSH: (3.94 +/- 1.52) IU/L, INH B: (162.49 +/- 78.38) ng/L, P < 0.01] and fertile volunteers [FSH: (4.27 +/- 2.84) IU/L, INH B: (228.49 +/- 110.68) ng/L, P < 0.01]. Mean serum inhibin B were significantly higher in patients with nonobstructive azoospermia who had spermatozoa on TESE than in those in whom no spermatozoa was found on TESE [INHB: (90.31 +/- 72.18) ng/L vs (19.54 +/- 20.38) ng/L, r = 0.528, P < 0.01], but mean FSH levels did not have similar predictive power (P>> 0.05).

CONCLUSIONS

Serum INH B level seems to be more accurate than serum FSH in the prediction of presence of testicular spermatozoa in patients with nonobstructive azoospermia. Serum inhibin B determination may be substitute of TESE as a diagnostic index.

OBJECTIVE

To investigate the relationship between pathological alterations of spermatogenic impairment in seminiferous tubules and serum inhibin B concentration in patients with azoospermia and to verify the significance of INH B in evaluating spermatogenesis.

METHODS

Eighty-three cases of azoospermia underwent testicular biopsy for the purpose of diagnosis. In accordance with the pathological alterations of spermatogenesis in seminiferous tubules, the samples were divided into four groups: Sertoli cell-only syndrome (n = 21); hypospermatogenesis (n = 20); maturation arrest (n = 24) and almost normal spermatogenesis (n = 18). Serum INHB and FSH, LH, T concentrations were tested before testicular biopsy for each patient respectively.

RESULTS

The INHB levels were (20. 85 +/- 18.78) pg/ml, (67.25 +/- 40.98) pg/ml, (73.63 +/- 25.54) pg/ml and (149.48 +/- 27.92) pg/ml in the above four groups, respectively. There was no significant statistical difference in the level of serum INH B between maturation arrest and hypospermatogenesis groups (P>> 0.05), and there was a very significant difference in almost normal spermatogenesis group and the other three groups, respectively (P < 0.001). There was no significant difference in the concentration of serum FSH when maturation arrest group compared with spermatogenesis group (P>> 0.05), whereas between the other two groups and between each of them and maturation arrest or almost normal spermatogenesis there was a very significant difference in the level of serum FSH (P < 0.05); The concentrations of LH and T were not significantly different among the four groups (P>> 0.05).

CONCLUSIONS

Serum INHB concentration was decreased when spermatogenesis got impaired. It dropped the most markedly in Sertoli cell-only syndrome group. INH B reflects directly the spermatogenic function in seminiferous tubules of the testis. Therefore, it could be considered valuable for spermatogenesis and potential fertility in patients with azoospermia.

(<em>b</em>)Introduction</<em>b</em>): HAE is a very de<em>b</em>ilitating disease that causes significant distress for patients not only during an acute attack <em>b</em>ut also constant fear for a su<em>b</em>sequent attack. It is important to address long-term prophylactic (LTP) therapy to prevent attacks, decrease mor<em>b</em>idity and increase quality of life. When discussing LTP, the drug <em>b</em>urden, convenience and efficacy must <em>b</em>e taken into account.(<em>b</em>)Areas covered</<em>b</em>): We review the literature and the different phases of clinical trials leading up to approval <em>b</em>y the US FDA of su<em>b</em>cutaneous highly concentrated C1-Inhi<em>b</em>itor (SC-C1-<em>INH</em>), called Haegarda. The dose approved is of 60 IU/kg twice weekly showing significant improvement in reduction of attacks and need for on-demand therapy for attacks with minimal side effects.(<em>b</em>)Expert opinion</<em>b</em>): SC-C1-<em>INH</em> has added significantly to the armamentarium of physicians that treat HAE. The a<em>b</em>ility to achieve a steady state of C1-<em>INH</em> a<em>b</em>ove 40% function is key to the success of the drug. The drug <em>b</em>urden is a SC injection twice a week that exceeds the newly approved lanadeluma<em>b</em>. The <em>b</em>enefit may <em>b</em>e that the protein that is deficient in HAE is replaced and with this the complement, fi<em>b</em>rinolytic, coagulation pathways and contact system are also regulated; however, evidence that this is of <em>b</em>enefit is still lacking.

Members of two Australian families with type A Hereditary Angioedema (HAE), having affected individuals in three generations, were typed for a large number of genetic marker systems in a search for close linkage with the locus controlling C1 inhibitor (C1 inh). The evidence from both families indicated lack of close linkage with HLA or with the loci for Bf and GLO on chromosome 6. Very close linkage was also excluded between the locus for C1 inh and the loci for 6PGD, PGM1 and MNSs. The other markers were not informative, but data on all systems showing variation are reported. The publication of similar data for other kindreds will help to determine lod scores for the probability of linkage between the C1 inh locus and loci controlling common protein polymorphisms. Linkage studies of this kind could establish whether the loci controlling type A and B HAE are identical or separate.

OBJECTIVE

Low complement factor C4 is usually considered a valuable screening tool for patients with the potentially life-threatening hereditary angioedema with C1-inhibitor (C1-INH) deficiency (C1-INH-HAE). However, there are patients with C1-INH-HAE presenting with normal C4 levels. This means, that C1-INH-HAE may potentially be overlooked, if screening is performed only by measurement of C4. It has been suggested that measurement of C4 activation products is better suited to avoid false negative results. Our aim was to investigate whether total antigenic C4 or non-functional C4c is a better measure of the increased C4 activation in C1-INH-HAE patients.

METHODS

Two different monoclonal antibodies (mAb) to human C4 were produced: one had specificity for the β-chain of C4 and would thus react with both functional and non-functional C4, and the other was developed against the factor I cleavage site on the α3-domain of C4 and was thus specific for activated, non-functional C4c. With these mAb we investigated plasma from 19 Danish C1-INH-HAE patients in three different enzyme-linked immunosorbent assays (ELISAs): a total antigenic C4 assay, a functional C4 assay and an assay measuring non-functional C4c.

RESULTS

The amount of total antigenic C4 varied considerably between patients and 2 patients had total antigenic C4 levels in the normal area. Functional C4 was low in all C1-INH-HAE patients. A C4c/C4 ratio showed that around half the C4 measured in patients was non-functional and captured all C1-INH-HAE patients.

CONCLUSIONS

This study shows that the C4c/C4 ratio seems to be a better diagnostic measure than total antigenic C4 alone. Our findings underline that screening with total antigenic C4 implies a risk of overlooking C1-INH-HAE patients.

Since coccidioidal granulomas are histologically indistinguishable from tuberculous granulomas, a long course of isonicotinic acid hydrazide therapy was tried experimentally in three cases of coccidioidomycosis, with good results. In two cases the disease was far advanced and prognosis poor before INH therapy was begun. In one case the disease was mild and symptoms abated after a short course of small doses of INH. It recurred when INH therapy was discontinued, and again resolved when larger doses of INH were given over a longer period.INH seemed to have an effect on appetite also, although the patients were taking B-complex vitamins both before and during INH treatment. The three patients ill with coccidioidomycosis averaged a weight gain of four and a half pounds a month during the period of INH therapy. Six well persons who were underweight and lacked appetite were given INH without other drugs, and they then had an increase in appetite and in weight.

UNASSIGNED

To explore the clinical effect of testicular artery-sparing microscopic varicocelectomy (MV) in combination with Qilin Pills (QL) in the treatment of bilateral varicocele-induced oligoasthenospermia.

METHODS

Sixty patients with bilateral varicocele-induced oligoasthenospermia were randomly assigned to receive MV (n=30) or MV+QL (n=30), those in the latter group treated with oral QL for 12 weeks postoperatively. At 4, 8, and 12 weeks after operation, we compared the semen volume, sperm concentration, sperm motility, the levels of serum Inh B, luteinizing hormone (LH) and total testosterone (TT), and the testosterone secretion index (TSI) between the two groups.

RESULTS

After surgery, all the patients showed disappearance of varicocele symptoms, remarkably improved semen volume, sperm concentration, sperm motility, serum Inh B and TT levels, TSI, decreased LH and FSH (P<0.01). At 12 weeks after treatment, statistically significant differences were found between the MV and MV+QL groups in Inh B (138.96±22.26 vs 129.54±22.23) ng/L, LH (3.17±0.12 vs 3.59±0.11) IU/L, TT (13.98±3.02 vs 12.68±3.12) nmol/L, and TSI (4.41±0.53 vs 3.53±0.51) nmol/ IU (P<0.05). The pregnancy rate was significantly higher in the MV+QL than in the MV group (73.4% vs 36.6%, P<0.05).

CONCLUSIONS

Testicular artery-sparing microscopic varicocelectomy combined with Qilin Pills is an effective strategy for the treatment of bilateral varicocele-induced oligoasthenospermia by significantly improving the semen quality of the patient.

Using indirect immunofluorescence, three regulatory proteins of the complement system, C1-inhibitor (C1-INH), C3b inactivator (C3bINA) and beta 1H globulin have been detected in the glomeruli of patients with glomerulonephritis associated with complement activation. C1-INH and beta 1H were found frequently but C3bINA was rarely detected (only 14 biopsies). beta 1H globulin and C3b inactivator which modulate C3b activity were never found in the absence of C3, but C1-INH was sometimes found in the absence of C1s. The patterns for staining for C1s and C1-INH, C3 and beta 1H were almost identical suggesting that the regulatory proteins are binding to the proteins they regulate, as has been demonstrated in vitro. Thus, in tissues undergoing complement-mediated tissue damage, the extent of complement activation is controlled by the normal regulatory mechanisms.