The present study was aimed at fingerprinting pharmacoproteomic alterations of the Mycobacterium tuberculosis H37Rv strain induced by antitubercular drugs isoniazid (INH), ethambutol (EMB), and SQ109 [N-geranyl-N'-(2-adamantyl)ethane-1,2-diamine, a novel 1,2-diamine-based EMB analog], providing new understanding of pharmacoproteomic mechanisms of each and exploring new drug targets. The three drugs produced significant down-regulation of 13 proteins, including immunogenic ModD, Mpt64, with proteins from the Pro-Glu family being inhibited the most. Alternatively, the three drugs up-regulated 17 proteins, including secreted antigenic proteins ESAT-6 and CFP-10. Among these, ESAT-6 and AphC were most affected by INH, whereas EMB had the greatest effect on ESAT-6. All three drugs produced only moderate up-regulation of aerobic and iron metabolism proteins, i.e., electron transfer flavoprotein Fix A and Fix B, and ferritin-like protein BfrB, suggesting that the interruption of microbacterial energy metabolism is not a primary mechanism of action. INH suppressed ATP-dependent DNA/RNA helicase, but up-regulated beta-ketoacyl-acyl carrier protein synthase. These effects may contribute to its bactericidal effects. In contrast, EMB and SQ109 did just the opposite: these drugs up-regulated the helicase and down-regulated the synthase. For most of the H37Rv proteins, similar pharmacoproteomic patterns were found for both EMB and SQ109. None of the drugs significantly regulated expression of chaperonins GroES, GroEL2, and Dnak, suggesting that these drugs do not affect chaperone-mediated nascent polypeptide folding and sorting. The present study identified proteins directly modulated by the actions of INH, EMB, and SQ109 and distinguished INH activity from the diamine antitubercular compounds that inhibit M. tuberculosis H37Rv.

An HPLC method is described for measurement of plasma hydrazine (Hz) concentrations (CHz) at the same time as isoniazid (INH) levels (CINH). Study has been made of CHz during 2-5 after dose in healthy adults (A, n = 34), in adult pulmonary TB patients (B, n = 18) and in paediatric tuberculous meningitis patients (C, n = 25). Although the population has about equal proportions of 'slow' (52%) and 'fast' acetylators, in none of the groups could a correlation be shown between CHz levels or rates of Hz accumulation and any measure of acetylator type. Consequently Hz must be derived both from INH and from its metabolites during the first hours post-dose. For group A and ca. 70% of groups B and C a constant and maximal fraction of dose (ca. 0.6% for adults and 0.4% for paediatric patients) appeared as Hz at 4-5 h. For group B patients small pre-dose concentrations increased with duration of treatment. Four patients in group B showed the highest levels of CHz and rates of Hz accumulation some three times greater than the rest; all four had been identified as alcoholics and one showed evidence of hepatotoxicity at CHz (5 h) = 1.3% of dose. Amongst group C (9/25) episodes of high CHz greater than 0.5% of dose occurred during the first weeks of treatment and one developed CHz ca. 100 ng/ml = 1.3% of dose coincidentally with indications of hepatic damage.

OBJECTIVE

The Israel National Health Survey--World Mental Health Survey (INHS) was designed to collect data on (a) the prevalence of mental disorders; (b) the prevalence of impairments and disabilities; (c) chronic conditions, disability, physical health, health services utilization and out-of-pocket medical expenditure which might be associated with mental disorder; and (d) socioeconomic and demographic correlates of mental disorder. This paper presents an overview of the methods used in this survey.

METHODS

The INHS was a cross-sectional survey based on a representative sample of 5,000 adults, 21 years or older, from the general population of Israel. The Composite International Diagnostic Interview (WMH-CIDI) was administered in face-to-face interviews at the respondents' homes between May, 2003, and April, 2004, using computer assisted personal interview (CAPI) technology.

RESULTS

The overall response rate was 72.6%.

CONCLUSIONS

The methodology and the quality control procedures used have made the INHS database a unique source of information about the prevalence, disability burden and unmet health needs of people suffering from common mental disorders and substance disorders in Israel.

Serum levels of the complement proteins C3, C4, C1 inhibitor (C1 INH), factor I (C3b inactivator) and factor H (BIH) and plasma levels of cleavage products of C3 (C3c) and factor B were measured in 26 patients with acute pancreatitis. Breakdown of C3 occurred in 19 patients, as shown by a reduction in C3 level and the presence of C3c. C4 levels, however, did not fall and factor B breakdown products were not detected, thus suggesting that enzymatic cleavage of C3 occurred without significant involvement of either the early classical pathway or the alternative pathway. C1 INH and factor H both showed increases, presumably reflecting an acute phase response. Factor I showed an initial fall followed by a rise. There was no correlation between the presence or extent of C3 breakdown and the clinical condition of the patients. It is concluded that C3 cleavage in pancreatitis probably results from tryptic activity and that the measurement of complement components has no part to play in the management of the disease.

F11.2.32, a monoclonal antibody raised against HIV-1 protease (Kd = 5 nM), which inhibits proteolytic activity of the enzyme (K(inh) = 35(+/-3)nM), has been studied by crystallographic methods. The three-dimensional structure of the complex between the Fab fragment and a synthetic peptide, spanning residues 36 to 46 of the protease, has been determined at 2.2 A resolution, and that of the Fab in the free state has been determined at 2.6 A resolution. The refined model of the complex reveals ten well-ordered residues of the peptide (P36 to P45) bound in a hydrophobic cavity at the centre of the antigen-binding site. The peptide adopts a beta hairpin-like structure in which residues P38 to P42 form a type II beta-turn conformation. An intermolecular antiparallel beta-sheet is formed between the peptide and the CDR3-H loop of the antibody; additional polar interactions occur between main-chain atoms of the peptide and hydroxyl groups from tyrosine residues protruding from CDR1-L and CDR3-H. Three water molecules, located at the antigen-antibody interface, mediate polar interactions between the peptide and the most buried hypervariable loops, CDR3-L and CDR1-H. A comparison between the free and complexed Fab fragments shows that significant conformational changes occur in the long hypervariable regions, CDR1-L and CDR3-H, upon binding the peptide. The conformation of the bound peptide, which shows no overall structural similarity to the corresponding segment in HIV-1 protease, suggests that F11.2.32 might inhibit proteolysis by distorting the native structure of the enzyme.

Subnormal plasma fibronectin (Fn) levels are found in patients with severe abdominal infections (SAI). The repletion of Fn has been postulated to have therapeutic benefit by virtue of its opsonic, reticuloendothelial system (RES) stimulating effects. A controlled, prospective trial of Fn administration was performed in patients with SAI to assess its use as an adjunct to standard procedures of intensive care. Thirty-three SAI patients were given daily doses of 0.8 g of purified Fn on days 1-5 following admission to the ICU, whereas 34 control patients received no Fn. All patients received the clinical care, antibiotics, and pharmacologic agents appropriate to their individual needs. The admission status and laboratory profiles of the two patient groups (+ and -Fn) were comparable on admission to the study. No side effects of the Fn preparation were observed. As judged by subgroup averages, the Fn replacement regimen was effective in elevating Fn levels to within normal range from day 2 onwards, as measured by immunological and functional assays. The estimated intravascular recovery of Fn averaged 82% in those patients who survived, yet only 52% in the nonsurvivors. Ultimate hospital mortality was 9/33 (27.3%) in the +Fn group versus 13/34 (38.2%) in the -Fn group (p = 0.244, Fisher's exact test). Although ultimate mortality was not significantly changed by the administration of Fn, the Fn treated patients appeared to survive longer than did the control patients. This trend was confirmed through the analysis of expected survival curves (D = 3.12, 0.1 greater than p greater than 0.05). When compared to the survivors, the ultimate nonsurvivors entered the study with statistically higher group averages of bilirubin and creatinine concomitant with lower averages of Fn, antithrombin III, C4, C3, C3b-INH, and transferrin. These differences persisted throughout the 11-day monitoring period; differences between survivors and nonsurvivors with respect to platelets, plasminogen, B-1-H, alpha-2-macroglobulin, and prealbumin appeared during the same period. Dramatic differences between the +Fn and -Fn treatment groups were not seen. Other than Fn, the Fn recipients only developed higher levels of the acute phase reactants C4, C3b-INH, B-1-H and alpha-1-antitrypsin (p less than 0.05) than did their non-Fn treated counterparts. In the present study, we again found a highly significant pattern of correlations between the absolute levels as well as the changes of Fn and other plasma proteins.(ABSTRACT TRUNCATED AT 400 WORDS)

Host-directed therapies are gaining considera<em>b</em>le impetus <em>b</em>ecause of the emergence of drug-resistant strains of pathogens due to anti<em>b</em>iotic therapy. Therefore, there is an urgent need to exploit alternative and novel strategies directed at host molecules to successfully restrict infections. The C-type lectin receptor CLEC4E and Toll-like receptor TLR4 expressed <em>b</em>y host cells are among the first line of defense in encountering pathogens. Therefore, we exploited signaling of macrophages through CLEC4E in association with TLR4 agonists (C<su<em>b</em>)4</su<em>b</em>).T<su<em>b</em>)4</su<em>b</em>)) to control the growth of <i>Myco<em>b</em>acterium tu<em>b</em>erculosis</i> (<i>Mt<em>b</em></i>). We o<em>b</em>served significant improvement in host immunity and reduced <em>b</em>acterial load in the lungs of <i>Mt<em>b</em>-</i>infected mice and guinea pigs treated with C<su<em>b</em>)4</su<em>b</em>).T<su<em>b</em>)4</su<em>b</em>) agonists. Further, intracellular killing of <i>Mt<em>b</em></i> was achieved with a 10-fold lower dose of isoniazid or rifampicin in conjunction with C<su<em>b</em>)4</su<em>b</em>).T<su<em>b</em>)4</su<em>b</em>) than the drugs alone. C<su<em>b</em>)4</su<em>b</em>).T<su<em>b</em>)4</su<em>b</em>) activated MYD88, PtdIns3K, STAT1 and RELA/NFK<em>B</em>, increased lysosome <em>b</em>iogenesis, decreased <i>Il10</i> and <i>Il4</i> gene expression and enhanced macroautophagy/autophagy. Macrophages from autophagy-deficient (<i>atg5</i> knockout or <i><em>B</em>ecn1</i> knockdown) mice showed elevated survival of <i>Mt<em>b</em></i>. The present findings also unveiled the novel role of CLEC4E in inducing autophagy through MYD88, which is required for control of <i>Mt<em>b</em></i> growth. This study suggests a unique immunotherapeutic approach involving CLEC4E in conjunction with TLR4 to restrict the survival of <i>Mt<em>b</em></i> through autophagy. (<em>b</em>)A<em>b</em><em>b</em>reviations:</<em>b</em>) 3MA: 3 methyladenine; AO: acridine orange; Atg5: autophagy related 5; AVOs: acidic vesicular organelles; <em>B</em>ECN1: <em>b</em>eclin 1, autophagy related; <em>B</em>MDMs: <em>b</em>one marrow derived macrophages; <em>b</em>w: <em>b</em>ody weight; C<su<em>b</em>)4</su<em>b</em>).T<su<em>b</em>)4</su<em>b</em>): agonists of CLEC4E (C<su<em>b</em>)4</su<em>b</em>)/TD<em>B</em>) and TLR4 (T<su<em>b</em>)4</su<em>b</em>)/ultra-pure-LPS); CFU: colony forming unit; CLEC4E/Mincle: C-type lectin domain family 4, mem<em>b</em>er e; CLR: c-type lectin receptor; <em>INH</em>: isoniazid; LAMP1: lysosomal-associated mem<em>b</em>rane protein 1; Mφ^C4.T4: <i>Mt<em>b</em>-</i>infected C<su<em>b</em>)4</su<em>b</em>).T<su<em>b</em>)4</su<em>b</em>) stimulated macrophages; MAP1LC3/LC3: microtu<em>b</em>ule-associated protein 1 light chain 3; MDC: monodansylcadaverine; MTOR: mechanistic target of rapamycin kinase; MYD88: myeloid differentiation primary response 88; NFK<em>B</em>: nuclear factor of kappa light polypeptide gene enhance in <em>B</em> cells; NLR: NOD (nucleotide-<em>b</em>inding oligomerization domain)-like receptors; PFA: paraformaldehyde; PPD: purified protein derivative; PtdIns3K: class III phosphatidylinositol 3-kinase; RELA: v-rel reticuloendotheliosis viral oncogene homolog A (avian); RIF: rifampicin; RLR: retinoic acid-induci<em>b</em>le gene-I-like receptors; TD<em>B</em>: trehalose-6,6´-di<em>b</em>ehenate; TLR4: toll-like receptor 4; Ultra-pure-LPS: ultra-pure lipopolysaccharide-EK; V-ATPase: vacuolar-type H⁺ ATPase.

OBJECTIVE

This study was designed to evaluate the toxic effects of Atrazine (ATZ) on the reproductive system of male rats.

METHODS

Male Sprague-Dawley rats were exposed to ATZ by gavage at dosages of 0, 38.5, 77, and 154 mg/kg bw/day for 30 d. The toxic effects of ATZ to rats were assessed through histopathologcal observation, spermatozoa quality evaluation, testicular marker enzyme indicators, antioxidant capacity and reproductive hormone levels.

RESULTS

Significant adverse effects on reproductive system were observed in rats exposed to ATZ at different dosages compared with 0 mg/kg group, including an irregular and disordered arrangement of the seminiferous epithelium in 154 mg/kg group; a decreased spermatozoa number and an increased spermatozoa abnormality rate in 77 and 154 mg/kg groups; decreased levels of acid phosphatase (ACP), alkaline phosphatase (AKP), lactic dehydrogenase (LDH), and succinate dehydrogenase (SDH) with the increasing of ATZ concentration; a decreased level of total antioxidant capacity (TAC) in a dose-dependent manner, and a decreased reduced glutathione (GSH) level and an increased malondialdehyde (MDA) content in 154 mg/kg group; and decreased serum levels of testosterone (T) and inhibin-B (INH-B) and an increased serum level of follicle stimulating hormone (FSH) in 77 and 154 mg/kg groups, and an increased serum level of luteinizing hormone (LH) in 154 mg/kg group.

CONCLUSIONS

These results suggested that relatively high doses of ATZ could exert reproductive toxicity of male rats.

Xenotransplantation may be a future alternative due to increased shortage of organs. Classical complement activation is central in hyperacute rejection in pig-to-human combinations. We investigated the effects of C1-inhibitor (C1-INH), a regulator of the complement and contact systems, on hyperacute rejection. Pig kidneys were perfused with fresh human blood to which either C1-INH (n = 6) or human serum albumin (n = 6) was added. The survival of the C1-INH perfused kidneys (mean 327 min) was significantly longer (p < 0.00001) than the controls (79 min). C1-INH substantially inhibited complement activation (C1rs-C1-INH complexes, C4bc, C3bc and terminal complement complex) (p < 0.001 for all) compared with the marked complement activation in the controls. No contact activation was found. Leukocytes and platelets were substantially activated (counts, myeloperoxidase, beta-thromboglobulin, thrombospondin, soluble P-selectin) in the control group, and this activation was markedly reduced by C1-INH (p < 0.02 for all). Immunohistochemistry showed less C1q, C3, TCC, IgG and fibrin deposition in the C1-INH group. C1-INH may be useful to attenuate hyperacute rejection, probably through inhibition of complement. The reduced activation of neutrophils and platelets may mainly be secondary to inhibition of complement.

Tuberculosis (TB) is highly endemic in India. The first-line anti-TB therapy (ATT) involving isoniazid (INH), rifampicin and pyrazinamide causes hepatotoxicity in approximately 11.5% of Indian patients. Studies have shown that ATT-induced hepatotoxicity is primarily due to oxidative stress caused by the drugs and metabolites. Herbal drugs with antioxidative properties have been tested in animal studies and clinical trials for the management of hepatotoxicity. The objective of this study was to investigate the role of curcumin (CUR), silymarin (SILY) and N-acetylcysteine (N-ACET) on hepatotoxicity by ATT drugs using an in vitro model of human hepatocellular carcinoma cell line (HepG2). HepG2 cells were treated with ATT drugs alone or along with CUR, SILY or N-ACET for a 48-h duration. The cells were monitored for viability, morphology, respiring mitochondria and cell cycle. Our results suggest that the presence of hepatoprotective drugs during treatment of HepG2 cells with ATT drugs lowers the hepatotoxic effect of the latter. This is observed in terms of (a) increased cell viability, (b) healthy-looking cell morphology as revealed by phase contrast microscopy, (c) active respiring cells as observed with confocal microscopy upon staining with a mitochondrial membrane-specific dye, MitoTracker(®) Red, and reduction in the sub-G(1) peak in cell cycle analysis by flow cytometry. Our results suggest that these hepatoprotective drugs need to be further explored as potential adjuvant therapy along with ATT drugs.

Rifampicin and pyrazinamide constitute an important part of drug regimens advocated for tuberculosis therapy, along with INH and streptomycin/ethambutol. The International Union against Tuberculosis and Lung Disease has also suggested the inclusion of these two drugs as part of short course chemotherapy for tuberculosis. Hence this study was undertaken to evaluate the influence of pyrazinamide on rifampicin kinetics when the two are given together. In a randomized, cross-over, single dose study, 16 patients with untreated pulmonary tuberculosis, after an overnight fast, were administered either rifampicin 450 mg + INH 300 mg (study A) or rifampicin 450 mg + INH 300 mg+pyrazinamide 1500 mg (study B). Blood samples were collected for serum rifampicin estimation at 0, 0.5, 2, 4, 6 and 8 h. Various pharmacokinetic parameters (Cmax, Tmax, t1/2, Kel, area under plasma concentration time curve (AUC), Vd & Cpl of rifampicin) were calculated. It was observed that rifampicin concentration in study A in contrast to study B was significantly higher at 6 h (P < 0.01) and 8 h (P < 0.05), while there were no significant differences in serum rifampicin concentration at 0.5, 2 and 4 h. A significant difference was also observed in AUC and Cpl. In study A, AUC was higher (P < 0.05), while Cpl was lower (P < 0.02), than in study B. From the above data it appears that on concomitant administration of pyrazinamide in patients on rifampicin therapy, AUC of rifampicin is decreased while its clearance is increased.

Double-stranded (ds) DNA, DNA- or RNA-associated nucleoproteins are the primary autoimmune targets in SLE, yet their relative inability to trigger similar autoimmune responses in experimental animals has fascinated scientists for decades. While many cellular proteins bind non-specifically negatively charged nucleic acids, it was discovered only recently that several intracellular proteins are involved directly in innate recognition of exogenous DNA or RNA, or cytosol-residing DNA or RNA viruses. Thus, endosomal Toll-like receptors (TLR) mediate responses to double-stranded RNA (TLR-3), single-stranded RNA (TLR-7/8) or unmethylated bacterial cytosine (phosphodiester) guanine (CpG)-DNA (TLR-9), while DNA-dependent activator of IRFs/Z-DNA binding protein 1 (DAI/ZBP1), haematopoietic IFN-inducible nuclear protein-200 (p202), absent in melanoma 2 (AIM2), RNA polymerase III, retinoic acid-inducible gene-I (RIG-I) and melanoma differentiation-associated gene 5 (MDA5) mediate responses to cytosolic dsDNA or dsRNA, respectively. TLR-induced responses are more robust than those induced by cytosolic DNA- or RNA- sensors, the later usually being limited to interferon regulatory factor 3 (IRF3)-dependent type I interferon (IFN) induction and nuclear factor (NF)-kappaB activation. Interestingly, AIM2 is not capable of inducing type I IFN, but rather plays a role in caspase I activation. DNA- or RNA-like synthetic inhibitory oligonucleotides (INH-ODN) have been developed that antagonize TLR-7- and/or TLR-9-induced activation in autoimmune B cells and in type I IFN-producing dendritic cells at low nanomolar concentrations. It is not known whether these INH-ODNs have any agonistic or antagonistic effects on cytosolic DNA or RNA sensors. While this remains to be determined in the future, in vivo studies have already shown their potential for preventing spontaneous lupus in various animal models of lupus. Several groups are exploring the possibility of translating these INH-ODNs into human therapeutics for treating SLE and bacterial DNA-induced sepsis.

Activation of TLR7 and TLR9 by endogenous RNA- or DNA-containing ligands, respectively, is thought to contribute to the complicated pathophysiology of systemic lupus erythematosus (SLE). These ligands induce the release of type-I interferons by plasmacytoid dendritic cells and autoreactive antibodies by B-cells, both responses being key events in perpetuating SLE. We recently described the development of inhibitory oligonucleotides (INH-ODN), which are characterized by a phosphorothioate backbone, a CC(T)XXX3-5GGG motif and a chemical modification of the G-quartet to avoid the formation of higher order structures via intermolecular G-tetrads. These INH-ODNs were equally or significantly more efficient to impair TLR7- and TLR9-stimulated murine B-cells, macrophages, conventional and plasmacytoid dendritic cells than the parent INH-ODN 2088, which lacks G-modification. Here, we evaluate the inhibitory/therapeutic potential of our set of G-modified INH-ODN on human immune cells. We report the novel finding that G-modified INH-ODNs efficiently inhibited the release of IFN-α by PBMC stimulated either with the TLR7-ligand oligoribonucleotide (ORN) 22075 or the TLR9-ligand CpG-ODN 2216. G-modification of INH-ODNs significantly improved inhibition of IL-6 release by PBMCs and purified human B-cells stimulated with the TLR7-ligand imiquimod or the TLR9-ligand CpG-ODN 2006. Furthermore, inhibition of B-cell activation analyzed by expression of activation markers and intracellular ATP content was significantly improved by G-modification. As observed with murine B-cells, high concentrations of INH-ODN 2088 but not of G-modified INH-ODNs stimulated IL-6 secretion by PBMCs in the absence of TLR-ligands thus limiting its blocking efficacy. In summary, G-modification of INH-ODNs improved their ability to impair TLR7- and TLR9-mediated signaling in those human immune cells which are considered as crucial in the pathophysiology of SLE.

Current anti-angiogenic therapy for cancer is based mainly on inhibition of the vascular endothelial growth factor pathway. However, due to the transient and only modest benefit from such therapy, additional approaches are needed. Deregulation of microRNAs (miRNAs) has been demonstrated to be involved in tumor angiogenesis and offers opportunities for a new therapeutic approach. However, effective miRNA-delivery systems are needed for such approaches to be successful. In this study, miRNA profiling of patient data sets, along with in vitro and in vivo experiments, revealed that miR-204-5p could promote angiogenesis in ovarian tumors through THBS1. By binding with scavenger receptor class B type 1 (SCARBinhibitor (miR-204-5p-inh) to tumor sites to suppress tumor growth. These results offer a new understanding of miR-204-5p in regulating tumor angiogenesis.

Isoniazid (INH) is associated with one of the highest incidences of idiosyncratic drug-induced liver failure of any commonly prescribed drug. The mechanism of this liver injury remains uncertain, and a valid animal model would greatly facilitate mechanistic studies. Most studies of INH-induced liver toxicity have been acute studies performed in rats with high doses of the drug, and this is very different from the idiosyncratic liver injury that occurs in humans. It has previously been demonstrated that covalent binding of INH in the liver of mice is greater than in rats and more like that in humans. Therefore, mice should be a better species in which to develop an animal model of INH-induced liver injury. Treatment of Cbl-b(-/-) and PD1(-/-) mice, which have impaired immune tolerance, resulted in greater injury than their C57BL/6 background, but not liver failure. This suggested that the injury was mediated by the adaptive immune system; however, Rag(-/-) mice, which do not have competent T- and B-cells, sustained more liver injury than C57BL/6 wild-type mice. This suggested that the adaptive immune system also played a protective role. INH treatment also led to a decrease in the inflammatory cytokines IL-1α and IL-12, which suggests that the drug may have immunosuppressive properties. In short, a mouse model was developed of INH-induced liver injury in which the immune system appears to play a both protective and pathogenic role, but this study was unable to develop a model of INH-induced liver failure.

BACKGROUND

Inhibin B (Inh B) is produced by pre-antral and early antral follicles whereas estradiol (E(2)) is a product of follicles undergoing antrum formation. This temporal distinction is evident in the patterns of Inh B and E(2) release earlier and later during the follicular phase of the menstrual cycle, respectively. However, in previous studies of women with polycystic ovary syndrome (PCOS) and normal controls, release of these granulosa cell (GC) products appears to be simultaneous in response to FSH stimulation. In order to reconcile these disparate findings, we conducted dose-response studies in both PCOS women and normal controls to determine whether GC product responses were due to the amount of FSH administered. In addition, we compared FSH-stimulated responses in PCOS women at various stages of recovery following ovarian suppression with a long-acting GnRH agonist to examine whether Inh B and E(2) responses reflected the level of ovarian follicle activity (i.e. circulating E(2) levels).

METHODS

Women with PCOS, 18-35 years (n = 23), and normal ovulatory controls, 18-35 years (n = 10) were recruited for study. Dose-responses were assessed over 24 h following intravenous administration of 0 (saline), 37.5, 75 and 150 IU of recombinant human FSH (r-hFSH) in PCOS and normal women. In addition, E(2) and Inh B responses to 150 IU of r-hFSH were assessed at baseline and 4, 6 and 8 weeks following suppression of ovarian steroidogenesis by a long-acting GnRH agonist in PCOS women.

RESULTS

In PCOS women and normal controls, serum Inh B and E(2) exhibit similar and simultaneous dose-responsiveness to FSH stimulation. During recovery from ovarian suppression, basal and stimulated Inh B release appear to be restored earlier than that of E(2) in PCOS women.

CONCLUSIONS

These findings are consistent with the notion that, in PCOS women, the level of ovarian follicle activity largely determines the earlier release of Inh B compared with E(2).

BACKGROUND

Hereditary angioedema (HAE) is caused by mutations in the C1inh gene, leading to dysfunction of the C1-esterase inhibitor (C1-INH). C1-INH interacts with MASP-1 and MASP-2 proteases, participating in the mannan-binding lectin (MBL) pathway of complement activation. The aim of the study was to investigate the contribution of possible changes in MBL/MASP-2 complex activity and Helicobacter pylori, hepatitis B virus (HBV), and hepatitis C virus (HCV) infections to the severity and frequency of clinical symptoms of HAE.

METHODS

The study was performed in 65 patients with HAE and 113 healthy persons. The parameters measured were C1-INH, C4, MBL concentration and MBL/MASP-2 complex activity, and serological markers of H. pylori, HBV, and HCV infection. Scores for the frequency and severity of HAE symptoms were determined.

RESULTS

HAE scores were significantly higher in patients whose C1-INH activity did not exceed 10% than in patients with activity of 10-52% (p=0.016). No significant differences were found in the median levels of MBL concentration and MBL/MASP-2 complex activity between patients and the control group. There was a slight association between contact with H. pylori in patients and HAE symptom score (p=0.052, not significant). Adult patients showed a 2.6-times higher frequency of anti-HBc than the general population. HBV DNA was negative in anti-HBc(+) patients.

CONCLUSIONS

These results suggest that the MBL complement activation pathway itself does not contribute to the frequency of angioedema attacks. Infections with H. pylori and HBV may slightly influence the disease score (not significant).

Inhibins (INH) are dimeric glycoproteins, composed of an alpha-subunit (INH-alpha) and one of two possible beta-subunits (INH-betaA or -betaB), with substantial roles in human reproduction and in endocrine-responsive tumors. The aims of this study were to determine the frequency and tissue distribution of INH-alpha, -betaA and -betaB in normal and malignant endometria. Samples were obtained from normal (n=46), atrophic (n=8) and endometrioid carcinoma tissue (EC; G1=93; G2=32; G3=14). INH-alpha was significantly higher in normal compared to malignant endometrial tissue, showing a cyclical variation throughout the menstrual cycle. EC G3 did not express this subunit. INH-betaA and -betaB showed specific staining reactions within the tumor cells. The highest intensity of INH-betaA was observed in the normal secretory phase compared to adenocarcinomas (p<0.05). For INH-betaB, the significantly highest expression was noted in EC G3 compared to EC G2 (p<0.05) and atrophic endometrial tissue. In conclusion, INH-alpha, -betaA and -betaB were immunolabeled in normal and malignant endometria. INH-alpha was expressed in a declining relationship in the transition from normal to tumor tissue, suggesting a tumor suppressive function in EC. A high expression of INH-betaB was observed in EC G3 compared to G2, suggesting an important role in the progression of endometrial carcinogenesis. However, the utilization of these subunits as specific tumor markers still remains unclear.

BACKGROUND

Duration of treatment in tuberculosis of spine has always been debatable in the absence of marker of healing. The objective of the study was to evaluate the efficacy of extended DOTS regimen (2 months of intensive phase and 6 months of continuation phase) as recommended by WHO, by using MRI observations as the healing marker.

METHODS

51 (Group A -28 prospective and Group B- 23 retrospective) patients of spine TB with mean age of 26.8 years (range 15-54 years) diagnosed clinico radiologically/imaging (n=36), histopathology or by PCR (n=15) were enrolled for the study. They were treated by extended DOTS regimen (2 months of HRZE and 6 months of HR) administered alternate day. The serial blood investigations and X-rays were done every 2 months. Contrast MRI was done at the end of 8 months and healing changes were recorded. Criteria of healing on the basis of MRI being: complete resolution of pre and paravertebral collections, resolution of marrow edema of vertebral body (VB), replacement of marrow edema by fat or by calcification suggested by iso- intense T1 and T2 weighted images in contrast enhanced MRI. Patients with non healed status, but, responding lesion on MRI after 8 months of treatment were continued on INH and rifampicin alternate day and contrast MRI was done subsequently at 12 months and 18 months till the healed status was achieved .

RESULTS

9 patients had paraplegia and required surgical intervention out of which 1 did not recover neurologically. All patients have completed 8 months of extended DOTS regimen, n=18 achieved healed status and duration of treatment was extended in rest (n=33) 5 were declared healed after 12 months, 8 after 18 months and one after 36 months of treatment, thus 32 were declared healed at varying periods.

CONCLUSIONS

35.2% patients demonstrate MRI based healed vertebral lesion at the end of 8 months of extended category 1 DOTS regimen. It is unscientific to stop the ATT by fixed time frame and MRI evaluation of the patients is required after 8 months of ATT and subsequently to decide for the continuation stoppage of treatment.

C1q, C1s and C1 Inh synthesized and secreted by human monocytes were characterized by SDS-PAGE. C1q is formed of three chains A (Mr approximately 35 000), B (Mr approximately 33 000) and C (Mr approximately 25 000) which are associated in two subunits A-B and C-C. It appears identical to C1q purified from plasma. C1s is secreted as a non-activated, monocatenar protein of Mr approximately 87 000 identical to proenzymic C1s from plasma. Secreted C1 Inh (Mr approximately 100 000) has a slightly higher Mr than purified plasmatic C1 Inh. Monensin treatment of the cells favours the intracytoplasmic accumulation of products at various glycosylation stages.

Mononuclear phagocytes release several factors involved in host defense and inflammation. Of these, interleukin-1 (IL-1) has multiple biological activities which are controlled at different levels including modulation of gene expression, protein synthesis or secretion, and interaction with inhibitors. We have investigated the production of IL-1 alpha and beta as well as the production of a specific IL-1 inhibitor (IL-1 INH) during the in vitro maturation of human monocyte-macrophages. Highly purified monocytes isolated by counterflow centrifugal elutriation were cultured up to six weeks, producing high levels of IL-1 alpha and beta during the first week of culture. Shortly after the first week bioactivity of IL-1 decreased, preceding a decrease of IL-1 immunoreactivity. In contrast, IL-1 inhibitory activity reached a peak during the third week and remained detectable up to six weeks. Granulocyte-monocyte-colony-stimulating factor GM-CSF increased the production of IL-1 INH by approximately 20%, but did not affect IL-1 production. The IL-1 INH, apparent molecular weight approximately 23 kD, blocks the binding of [125I]IL-1 alpha to its receptor. The balance between the production of IL-1 and its antagonist may be important for the regulation of the immune response and chronic inflammation during pathological processes.

589 isolates of anaerobic bacteria, including 127 isolates of Bacterioides fracilis, were tested against 23 antimicrobial agents. At their usually achievable serum levels, carbenicillin, ticarcillin, mezlocillin and piperacillin; cefoxitin, minocycline, doxycycline, clindamycin, chloramphenicol and rifampin were effective against more than 80% of B. fragilis isolates. At usual therapeutic doses, penicillin (PEN), ampicillin (AMP), cyclacillin (CYC), cephalothin (CT), cefazolin (CZ), cephradine (CPD), cefamandole (CMD), cefaclor (CCL), cefuroxime (CRX), spectinomycin (SPT), tetracycline (TET), ethambutol (EMB) and isoniazid (INH) were ineffective against B. Fragilis. However, high and clinically tolerated doses of PEN, CMD and SPT inhibited more than 80% of B. fragilis isolates, but AMP, CYC, CT, CZ, CPD, CCL and CRX, TET, EMB and INH remained totally ineffective against B. fragilis. The therapeutic merits of these antibiotics are discussed.

New therapeutic options have modified the natural history and health care costs of multiple sclerosis (MS). An epidemiological 25 years-long model-based cost-utility analysis was performed following the Italian National Health Service (INHS) and societal perspectives to compare costs and quality-adjusted life years of treatment with Interferon beta-1b (IFNB-1b) from diagnosis of clinically isolated syndrome (CIS) versus treating at subsequent conversion to clinically definite MS (CDMS). Among patients treated (untreated) with IFNB-1b from CIS diagnosis, 40,420 (43,700) converted to CDMS after 25 years; the estimated cumulative probability of converting to CDMS during the first 3 years was 72.90% (84.94%) (P < 0.0001). Early treatment with IFNB-1b is highly cost-effective for the INHS (incremental cost-effectiveness ratio: Euros 2,574.94) and dominant from the societal viewpoint. Sensitivity analyses confirmed the base case findings. Early treatment with IFNB-1b delays conversion to CDMS in CIS patients and might be a "good value for money" health care programme.

The immunochemical properties, catabolic clearance, and organ distribution of highly purified radiolabeled human C1-INH were studied after treatment with glycosidases. The antigenicity and C1s inhibitory activity of C1-INH were unimpaired after removal of sialic acid residues (60%) or sialic acid and galactose residues (60 and 20%, respectively) with immobilized neuraminidase and beta-galactosidase. Treatment of C1-INH with neuraminidase was associated with increased cathodal migration of the glycoprotein. In vivo studies in the rabbit, however, showed that removal of sialic acid residues resulted in rapid blood clearance of C1-INH and enhanced localization of the asialo C1-INH in the liver. Subsequent removal of penultimate galactose residues returned the survival time in the circulation and organ distribution to near normal. Additionally, simultaneous infusion of asialo-C1-INH with asialo alpha 1-acid glycoprotein, a protein known to bind to hepatic asialoglycoprotein receptors, dramatically extended the intravascular survival time of asialo C1-INH. In agreement with the observations on other plasma glycoproteins, these results indicate that desialylation of C1-INH results in rapid clearance by hepatic asialoglycoprotein receptors.