<A<em>b</em>stractText>Drug resistant tu<em>b</em>erculosis (TB) has <em>b</em>ecome a persistent health threat in Ethiopia. In this respect, <em>b</em>aseline data are scarce in many parts of high TB <em>b</em>urden regions including the different zones of Ethiopia.</A<em>b</em>stractText><A<em>b</em>stractText>A total of 111 culture positive M. tu<em>b</em>erculosis isolates were recovered from TB patients and identified using region of difference (RD) 9 <em>b</em>ased polymerase chain reaction (PCR) and spoligotyping. Thereafter, their drug sensitivities to Rifampicin (RIF) and Isoniazid (<em>INH</em>) were evaluated using GenoType MTBDRplus assay.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>The result showed that 18.0% (20/111) of the isolates were resistant to either RIF or <em>INH</em>. Furthermore, 16.7 and 23.8% of the isolates from new and retreatment cases were resistant to any of the two anti-TB drugs, respectively. Multi-drug resistant (MDR) TB was detected on 1.8% (2/111) of all cases. Significantly higher frequencies of any drug resistance were o<em>b</em>served among Euro-American (EA) major lineage (χ²: 9.67; p = 0.046).</p><A<em>b</em>stractText>Considera<em>b</em>ly high proportion of drug resistant M. tu<em>b</em>erculosis strains was detected which could suggest a need for an increased effort to strengthen TB control program in the study area.</A<em>b</em>stractText>

Whereas guidelines recommend the routine use of natriuretic peptides (NPs) in heart failure (HF) care, the clinical relevance and prognostic potential of midregional pro-adrenomedullin (MR-proADM) is less well established. We aimed to compare the prognostic potential of MR-proADM after acute decompensation for systolic HF with that of N-terminal pro-B-type natriuretic peptide (NT-proBNP) and midregional pro-atrial NP (MR-proANP), to investigate the significance of high/rising MR-proADM, and to evaluate the incremental prognostic yield of repeat measurements.

The Interdisciplinary Network Heart Failure (INH) programme enrolled patients hospitalized for acute systolic HF and followed them for 18 months (100% complete). Of 1022 INH participants, 917 (68 ± 12 years, 28% female) who had biomaterials available were enrolled. High MR-proADM was associated with more impaired left ventricular function, higher comorbidity burden, lower doses of HF medications, and lower likelihood of left ventricular reverse remodelling. Compared with NPs, MR-proADM had superior prognostic significance (concordance index 0.72 for all-cause mortality), improved Cox regression models including NPs (P < 0.001), and was the only biomarker also predicting non-cardiac death (hazard ratio 1.8 vs. 1.0). In the setting of low NPs, patients with high MR-proADM experienced non-cardiac death more often. Six month MR-proADM enhanced models including baseline MR-proADM (P < 0.001) for prediction of all-cause death (net reclassification index: 0.48, 95% confidence interval 0.19-0.78).

MR-proADM was found to correlate with the global disease burden in HF and proved a potent prognostic indicator, capturing the risk for both cardiac and non-cardiac death. Serial MR-proADM measurements further enhanced risk assessment, thus facilitating substantial reclassification.

Bothrops lanceolatus snake venom causes systemic thrombotic syndrome but also local inflammation involving extensive oedema, pain, and haemorrhage. Systemic thrombotic syndrome may lead to fatal pulmonary embolism and myocardial and cerebral infarction. Here, we investigated the ability of B. lanceolatus venom to activate the Complement system (C) in order to improve the understanding of venom-induced local inflammation. Data presented show that B. lanceolatus venom is able to activate all C-pathways. In human serum, the venom strongly induced the generation of anaphylatoxins, such as C5a and C4a, and the Terminal Complement complex. The venom also induced cleavage of purified human components C3, C4, and C5, with the production of biologically active C5a. Furthermore, the venom enzymatically inactivated the soluble C-regulator and the C1-inhibitor (C1-INH), and significantly increased the expression of bound C-regulators, such as MCP and CD59, on the endothelial cell membrane. Our observations that B. lanceolatus venom activates the three Complement activation pathways, resulting in anaphylatoxins generation, may suggest that this could play an important role in local inflammatory reaction and systemic thrombosis caused by the venom. Inactivation of C1-INH, which is also an important inhibitor of several coagulation proteins, may also contribute to inflammation and thrombosis. Thus, further in vivo studies may support the idea that therapeutic management of systemic B. lanceolatus envenomation could include the use of Complement inhibitors as adjunct therapy.

<A<em>b</em>stractText>Hereditary angioedema (HAE) comprises HAE with C1-inhi<em>b</em>itor deficiency (C1-<em>INH</em>-HAE) and HAE with normal C1-<em>INH</em> activity (nl-C1-<em>INH</em>-HAE), due to mutations in factor XII (FXII-HAE), plasminogen (PLG-HAE), angiopoietin 1 (ANGPT1-HAE), kininogen 1 genes (KNG1-HAE) or angioedema of unknown origin (U-HAE). The Italian network for C1-<em>INH</em>-HAE (ITACA) created a registry including different forms of angioedema without wheals.</A<em>b</em>stractText><A<em>b</em>stractText>We analyzed clinical and la<em>b</em>oratory features of a cohort of Italian su<em>b</em>jects with nl-C1-<em>INH</em>-HAE followed <em>b</em>y ITACA to identify specific <em>b</em>iomarkers.</A<em>b</em>stractText><A<em>b</em>stractText>105 nl-C1-<em>INH</em>-HAE patients were studied. Plasma concentrations of cleaved high molecular weight kininogen (cHK), Vascular Endothelial Growth Factors (VEGFs), angiopoietins (Angs) and secreted phospholipase A2 enzymes (sPLA2) were evaluated.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>We identified 43 FXII-HAE patients, 58 U-HAE and 4 ANGPT1-HAE. We assessed a prevalence of 1:1.4 x 10⁶ for FXII-HAE and 1:1.0 x 10⁶ for U-HAE. cHK levels in U-HAE patients were similar to controls in plasma collected using protease inhi<em>b</em>itors cocktail (PIC), <em>b</em>ut they significantly increased in a<em>b</em>sence of PIC. In FXII-HAE patients cHK levels, in a<em>b</em>sence of PIC, were significantly higher than in controls. We found a significant increase of VEGF-A, VEGF-C, Ang1 levels in U-HAE patients compared to controls. In FXII-HAE only VEGF-C levels were increased. Ang2 concentrations and sPLA2 activity were not modified. The levels of these mediators in ANGPT1-HAE patients were not altered.</p><A<em>b</em>stractText>Our results suggest that pathogenesis of FXII-, ANGPT1- and U-HAE moves through an un<em>b</em>alanced control of kallikrein activity, with <em>b</em>radykinin as most likely mediator. VEGFs and Ang1 participate in the pathophysiology of U-HAE increasing the <em>b</em>asal vascular permea<em>b</em>ility.</A<em>b</em>stractText>

Background: Hereditary angioedema (HAE) is a potentially fatal disorder resulting in recurrent attacks of severe swelling. It may be associated with a genetic deficiency of functional C1 inhibitor (C1-INH) or with normal C1-INH (HAEnCI). In families with HAEnCI, HAE-linked mutations in the F12, PLG, KNG1, ANGPT1, or MYOF genes have been identified. In many families with HAEnCI the genetic cause of the disease is currently unknown.

Objective: The aim of this study was to identify a novel disease-linked mutation for HAEnCI.

Methods: Methods comprised whole exome sequencing (WES), Sanger sequencing analysis, pedigree analysis, bioinformatical analysis of the mutation, and biochemical analysis of parameters of the kallikrein-kinin (contact) system.

Results: By performing WES on a multi-generation family with HAEnCI we identified the HS3ST6 mutation c.430A>T (p.Thr144Ser) in all three affected family members that were sequenced. This gene encodes the heparan sulfate glucosamine 3-O-sulfotransferase 6 (3-OST-6) which is involved in the last step of heparan sulfate biosynthesis. The p.Thr144Ser mutation is likely to affect the interaction between two beta sheets stabilizing the active center of the 3-OST-6 protein.

Conclusions: We conclude that mutant 3-OST-6 fails to transfer sulfo groups to the 3-OH position of heparan sulfate, resulting in incomplete heparan sulfate biosynthesis. This is likely to affect cell surface interactions of key players in angioedema formation and is a novel mechanism for disease development.

Keywords: 3-OST-6; HAEnCI; HS3ST6; Hereditary angioedema; heparan sulfate glucosamine 3-O-sulfotransferase 6; normal C1-INH.

<p><div>(<em>b</em>)RATIONALE</<em>b</em>)</div>Despite therapeutic progress in treating cystic fi<em>b</em>rosis (CF) airway disease, airway inflammation with associated mucociliary dysfunction remains largely unaddressed. Inflammation reduces the activity of apically expressed large conductance, Ca²⁺-activated and voltage-dependent K⁺ channels (BK), critical for mucociliary function in the a<em>b</em>sence of CFTR.</p><A<em>b</em>stractText>Losartan's anti-inflammatory effectiveness to rescue BK activity and there<em>b</em>y mucociliary function was tested in vitro using primary, fully re-differentiated human airway epithelial cells homozygous for F508del and in vivo using a previously validated, now expanded pharmacological sheep model of CF- and inflammation-associated mucociliary dysfunction.</A<em>b</em>stractText><p><div>(<em>b</em>)MEASUREMENTS AND MAIN RESULTS</<em>b</em>)</div>Nasal scrapings from CF patients showed that neutrophilic inflammation correlated with reduced expression of LRRC26, the γ su<em>b</em>unit mandatory for BK function in the airways. TGF-β1, downstream of neutrophil elastase, decreased mucociliary parameters in vitro. These were rescued <em>b</em>y losartan at concentrations achieved <em>b</em>y ne<em>b</em>ulization in the airway and oral application in the <em>b</em>loodstream: BK dysfunction recovered acutely and over time (the latter via an increase in LRRC26 expression); ciliary <em>b</em>eat frequency and airway surface liquid (ASL) volume improved; and mucus hyperconcentration and cellular inflammation decreased. These effects did not depend on angiotensin receptor <em>b</em>lockade. Expanding on a validated and pu<em>b</em>lished nongenetic sheep model of CF, ewes <em>inh</em>aled CFTR<su<em>b</em>)<em>inh</em></su<em>b</em>)172 and neutrophil elastase for three days, which resulted in prolonged tracheal mucus velocity reduction, mucus hyperconcentration and increased TGF-β1. Ne<em>b</em>ulized losartan rescued <em>b</em>oth mucus transport and mucus hyperconcentration and reduced TGF-β1.</p><A<em>b</em>stractText>Losartan effectively reversed CF- and inflammation-associated mucociliary dysfunction, independent of its angiotensin receptor <em>b</em>lockade.</A<em>b</em>stractText>

Proteinase inhibitor I (Inh I) and proteinase inhibitor II (Inh II) from potato tubers are effective proteinase inhibitors of chymotrypsin and trypsin. Inh I and Inh II were shown to suppress irradiation-induced transformation in mouse embryo fibroblasts suggesting that they possess anticarcinogenic characteristics. We have previously demonstrated that Inh I and Inh II could effectively block UV irradiation-induced activation of transcription activator protein 1 (AP-1) in mouse JBInh I and Inh II on the expression and composition pattern of the AP-1 complex following stimulation by UV B (UVB) irradiation in the JBInh I and Inh II specifically inhibited UVB-induced AP-1, but not NFkappaB, activity in JBBoth Inh I and Inh II up-regulated AP-1 constituent proteins, JunD and Fra-2, and suppressed c-Jun and c-Fos expression and composition in bound AP-1 in response to UVB stimulation. This regulation of the AP-1 protein compositional pattern in response to Inh I or Inh II may be critical for the inhibition of UVB-induced AP-1 activity by these agents found in potatoes.

BACKGROUND

Infundibuloneurohypophysitis (INH) is reported to be a self-limiting inflammatory disease involving neurohypophysis. The authors experienced a case of INH presenting a large mass compressing the brain stem.

METHODS

The patient exhibited polyuria followed by left hemiparesis and dysarthria lasting a year. Magnetic resonance imaging showed a large sellar mass extending into the right cavernous sinus and prepontine cistern and compressing pons. Endocrinologically, diabetes insipidus was diagnosed and anterior pituitary function was almost normal. Microscopic examination of the surgical specimen obtained by a transsphenoidal route demonstrated diffuse infiltration of lymphoid cells with predominance of B cells over T cells and the granulation tissue. The patient underwent 40 Gy local radiation because of initial misinterpretation of histologic findings as malignant lymphoma and short-term corticostertoid administration.

CONCLUSIONS

The mass gradually shrank and the patient has become neurologically intact in 6 months. At this moment, 67 months after the onset, the patient is free from disease and has no other lesion. INH seems to be a clinical entity possessing a wide spectrum from infundibular tumorlet to an aggressive sellar mass trespassing on surrounding structures.

To compare outcomes of popliteal artery aneurysm (PAA) repair by endovascular treatment, great saphenous vein (GSV) bypass, and prosthetic bypass.Single center retrospective analysis of patients presenting PAA from 2000 to 2013. Patients were divided into endovascular treatment (group A); GSV bypass (group B); and prosthetic graft bypass (group C). Outcomes were technical success, perioperative mortality, and morbidity. Survival, primary and secondary patency, and freedom from reintervention rate were estimated. Differences in ankle-brachial index (ABI), in-hospital length of stay (InH-Los), red blood cell (RBC) transfusion, and limb loss were reported. Mean follow-up was 49 (median: 35; 1-145; SD 42) months.Sixty-seven patients were included; 25 in group A, 28 in group B, and 14 in group C. PAA was symptomatic in 23 (34%) cases. Technical success was 100%. No perioperative death occurred. Three (4.5%) perioperative complications were reported with no significant difference between groups (P = 0.866). Five-years estimated survival was 78%. Estimated 5-years primary patency for groups A, B, and C was 71%, 81%, and 69%, respectively (P = 0.19). Estimated 5-years secondary patency for groups A, B, and C was 88%, 85%, and 84% (P = 0.85). Estimated 5-years freedom from reintervention for groups A, B, and C was 62%, 84%, and 70%, respectively (P = 0.16). A significant difference between preoperative ABI versus postoperative ABI was observed (P = 0.001). InH-LoS was significantly shorter in group A (P < 0.001). RBC transfusions were required significantly less in group A when compared to group C (P = 0.045). Overall limb salvage was achieved in all but 1 patient.PAA repair has good early and long-term outcomes with different treatment options. Endovascular treatment was not inferior to surgical repair with a reduced InH-LoS and RBC transfusion. It can be successfully employed even in nonelective setting. A randomized controlled trial with long-term follow-up and appropriate patient inclusion criteria is necessary to compare these 3 treatment options.

Many pathogenic microorganisms express fibronectin-binding molecules that facilitate their adherence to the extracellular matrix and/or entry into mammalian cells. We have previously described a Borrelia recurrentis gene, cihC that encodes a 40-kDa surface receptor for both, fibronectin and the complement inhibitors C4bp and C1-Inh. We now provide evidence for the expression of a group of highly homologues surface proteins, termed FbpA, in three B. hermsii isolates and two tick-borne relapsing fever spirochetes, B. parkeri and B. turicatae. When expressed in Escherichia coli or B. burgdorferi, four out of five proteins were shown to selectively bind fibronectin, whereas none of five proteins were able to bind the human complement regulators, C4bp and C1-Inh. By applying deletion mutants of the B. hermsii fibronectin-binding proteins a putative high-affinity binding site for fibronectin was mapped to its central region. In addition, the fibronectin-binding proteins of B. hermsii were found to share sequence homology with BBK32 of the Lyme disease spirochete B. burgdorferi with similar function suggesting its involvement in persistence and/or virulence of relapsing fever spirochetes.

Bone tuberculosis (TB) is one of the most common extrapulmonary TB. Effective integration of chemotherapy and bone regeneration is an optimal solution for bone TB therapy. Herein, we produce a composite scaffold drug delivery system fabricated with an isoniazid conjugated star poly(lactide-co-glycolide) (PLGA-INHβ-TCP. The cytological assay indicated the composite system possesses good biocompatibility. The in vitro and in vivo drug release evaluations showed that the composite system can intactly release the pristine INH and maintain effective INH concentration in a controlled manner for more than 100 days, and achieve high localized drug concentration and low systemic drug concentration. The rabbit radius repair experiment testified the scaffold has good bone regeneration capacity. Our work demonstrate the composite system can simultaneously achieve localized long-term drug controlled release and bone regeneration, which provides a promising route for improved bone TB treatment.

The role of surface-bound type Ia group B Streptococcus (GBS) capsular polysaccharide in antibody-independent binding of C1 and activation of the classic complement pathway was investigated. In a radiolabeled bacterial-polymorphonuclear leukocyte (PMN) association assay, a measure of bacterial opsonization, preincubation of 3H-type Ia GBS with purified F(ab')2 to the organism blocked the association of the bacteria with PMN', and the inhibitory effect was dose dependent. The specificity of F(ab')2 blocking was shown after adsorption of F(ab')2 with type Ia polysaccharide-sensitized erythrocytes. Polysaccharide-adsorbed F(ab')2 had a 70% decrease in ability to block the association of bacteria with PMN. Evidence for the requirement of the capsular polysaccharide in classic complement pathway activation came from a C1 transfer assay with the use of neuraminidase-digested type Ia GBS. Neuraminidase digestion removed 80% of the terminal sialic acid residues from the native polysaccharide. These neuraminidase-digested organisms had a 72% decrease in binding and transfer of purified C1 compared with non-enzyme-treated organisms. Type Ia capsular polysaccharide bound to sheep erythrocytes promoted classic complement pathway-mediated hemolysis of the cells. The role of C1 inhibitor (INH) in modulation of C1 activation by the organisms was investigated. The possibility existed that the C1 INH could be bound by the bacteria, allowing C1 activation to occur in the fluid phase. The inhibitor was purified from human serum, and its activity was measured before and after incubation with type Ia GBS. The organisms had no effect on C1 INH activity. Thus surface-bound capsular polysaccharide of type Ia GBS mediates C1 binding and classic pathway activation, and this does not involve the C1 INH.

Hereditary angioedema (HAE) is a rare, potentially life-threatening disease that manifests as recurrent episodes of nonpruritic swelling that may affect the extremities, face, genitalia, gastrointestinal tract, and/or larynx. HAE is the result of a deficiency of functional C1-esterase inhibitor (C1-INH), a key regulator of the complement, coagulation, and kallikrein-kinin cascades. In HAE patients, overactivation of the kallikrein-kinin cascade results in excessive release of bradykinin, the mediator of the pain and swelling that is characteristic of HAE. Historically, treatment options for HAE have been limited, but newly approved and emerging therapies, such as C1-INH replacement products, a plasma kallikrein inhibitor, and a bradykinin B₂-receptor antagonist, appear to provide safe and effective relief for a significant proportion of patients with HAE. Because they may have therapeutic and practical advantages over existing HAE therapies, the new agents have the potential to improve the overall management of patients with HAE. This article reviews the results from recent clinical trials of these drugs and considers their role in clinical practice.

<p><div>(<em>b</em>)Introduction</<em>b</em>)</div>For prophylaxis of hereditary angioedema (HAE) attacks, replacement therapy with human C1-inhi<em>b</em>itor (C1-<em>INH</em>) treatment is approved and availa<em>b</em>le as intravenous [C1-<em>INH</em>(IV)] (Cinryze^®) and su<em>b</em>cutaneous [C1-<em>INH</em>(SC)] HAEGARDA^® preparations. In the a<em>b</em>sence of a head-to-head comparative study of the two treatment modalities, an indirect comparison of data from 2 independent <em>b</em>ut similar clinical trials was undertaken.</p><A<em>b</em>stractText>Two similar randomized, dou<em>b</em>le-<em>b</em>lind, place<em>b</em>o-controlled, crossover studies were identified which evaluated either C1-<em>INH</em>(SC) (COMPACT; NCT01912456; 16 weeks) or C1-<em>INH</em>(IV) (CHANGE; NCT01005888; 14 weeks) vs. place<em>b</em>o (on-demand treatment only) for routine prevention of HAE attacks. Individual patient data from each trial were used to conduct an indirect comparison of treatment effects. Attack reductions (a<em>b</em>solute and percent of mean/median num<em>b</em>er of monthly HAE attacks reduction over place<em>b</em>o) were compared <em>b</em>etween the two C1-<em>INH</em> formulations at approved/recommended doses: C1-<em>INH</em>(SC) 60 IU/kg twice weekly (n = 45) and 1000 U of C1-<em>INH</em>(IV) twice weekly (n = 22). Point estimates were adjusted using mixed and quantile regression models that controlled for study design.</A<em>b</em>stractText><p><div>(<em>b</em>)Results</<em>b</em>)</div>The a<em>b</em>solute mean monthly num<em>b</em>ers of HAE attack reductions were 3.6 (95% CI 2.9, 4.2) for C1-<em>INH</em>(SC) 60 IU/kg vs. place<em>b</em>o and 2.3 (1.4, 3.3) for C1-<em>INH</em>(IV) vs. place<em>b</em>o; <em>b</em>etween-product difference, 1.3 (0.1, 2.4; <i>P </i>= 0.034). The mean percent reduction in monthly attack rate was significantly greater with C1-<em>INH</em>(SC) as compared with C1-<em>INH</em>(IV) (84% vs. 51%; <i>P </i>< 0.001). The percentages of su<em>b</em>jects experiencing ≥ 50%, ≥ 70%, and ≥ 90% reductions in monthly HAE attack rates versus place<em>b</em>o were significantly higher with C1-<em>INH</em>(SC) 60 IU/kg as compared to C1-<em>INH</em>(IV) 1000 U (≥ 50% reduction: 91% vs. 50%, odds ratio [OR] = 10.33, <i>P </i>= 0.003; ≥ 70% reduction: 84% vs. 46%, OR = 6.19, <i>P</i> = 0.005; ≥ 90% reduction: 57% vs. 18%, OR = 6.04, <i>P</i> = 0.007).</p><A<em>b</em>stractText>Within the limitations of an indirect study comparison, this analysis suggests greater attack reduction with twice-weekly C1-<em>INH</em>(SC) 60 IU/kg as compared to twice-weekly C1-<em>INH</em>(IV) 1000 U for the routine prevention of HAE attacks.</A<em>b</em>stractText>

Dietzia spp. have broad potential applications in industries. However, genetic manipulation of these species is obstructed by their low transformation efficiency, which is in the range of 10(4)colony-forming units (CFU)μg(-1) exogenous DNA. In this study, both single-factor and orthogonal experiments were conducted to test the effects of competent cell concentration (parameter A), electroporation time (B), and field strength (C), as well as the concentrations of glycine (D), isonicotinic acid hydrazide (INH, E), Tween 80 (F), and penicillin G (G) on the electrotransformation efficiency of Dietzia sp. DQ12-45-1b. The order in which the parameters contributed to electrotransformation efficiency was C>D>F>G>B)A>E, suggesting that field strength, glycine, and Tween 80 each had a greater capability to increase electrotransformation efficiency than the other parameters tested. Using the optimized protocol deduced from the results of the orthogonal experiments, the electrotransformation efficiency of Dietzia sp. DQ12-45-1b reached 2.51×10(7)CFUμg(-1) plasmid. To the best of our knowledge, this is the highest transformation efficiency that has been reported for Dietzia bacteria to date. Thus, we present a method that combines the transformation factors to obtain the optimal transformation efficiency for a target bacterium.

Bridelia micrantha and Croton macrostachyus are medicinal plants used empirically in traditional medicine to treat epilepsy. In vivo mice model (maximal electroshock, strychnine, pentylenetetrazol, picrotoxin, isonicotinic hydrazide acid)-induced convulsions were used to evaluate the anticonvulsant activities of those plants. Diazepam-induced sleep was used for the evaluation of the sedative properties. B. micrantha protected 100, 80, 80, and 80% of mice against PIC, STR, PTZ and MES-induced seizures, respectively. C. macrostachyus at the doses 34 and 67 mg/kg protected 80, 80, 80 and 60% of mice from PIC, STR, PTZ and MES-induced seizures, respectively. B. micrantha and C. macrostachyus also delayed the onset to seizures in INH test. B. micrantha was more potent than C. macrostachyus in protecting mice against convulsions. The co-administration of the sub effective dose of the decoction of B. micrantha or C. macrostachyus with the sub effective dose of diazepam or clonazepam resulted in a synergistic effect. The decoctions of B. micrantha and C. macrostachyus also exerted sedative activity by increasing the total duration of sleep induced by diazepam and by reducing the latency time to sleep. The effect of the decoctions of B. micrantha and C. macrostachyus suggests the presence of anticonvulsant activities that might show efficacy against secondarily generalized tonic-clonic seizures and primary generalized seizures in humans.

The aim of our study was to determine whether the combination of an anticholinergic treatment with a beta 2-adrenergic medication is a more effective treatment for acute asthma attack than the two treatments individually. The association of salbutamol-ipratropium was compared to treatment with salbutamol and ipratropium alone. It was a prospective double-blind study in children with acute asthma attack, participating as outpatients. Their clinical history and characteristics of bronchial obstruction were recorded on a standard form. Afterwards, they were included in one of the three following study groups: group one, 100 micrograms/inh salbutamol; group two, 20 micrograms/inh ipratropium; group three, 100 micrograms/inh of salbutamol plus 20 micrograms/inh ipratropium. There were 40 patients in each group, with Tal score +/- 5 and PEF < 80% of the predicted value. They were evaluated at the beginning (0 min), and at 15, 30, 45, 60, 80, 100 and 120 min. Each patient was treated with two inhalations of the study medication and was then evaluated for variations in Tal score. The mean age was 7.3 years; Tal score was 5.6, 5.6 and 6.0 at 0 min (p>> 0.05). Decrease in Tal score after 15 min meant p < 0.01 for salbutamol-ipratropium and salbutamol vs. ipratropium. At 30 min p < 0.05 for salbutamol-ipratropium vs. salbutamol, and at 45 min p < 0.01 for salbutamol-ipratropium vs. salbutamol. PEF at 0 min was 70.9%, 71.3% and 68.6% (p>> 0.05) increasing after 15 min. At 30 min p < 0.05 for salbutamol-ipratropium vs. salbutamol, and p < 0.01 vs. ipratropium. At 45 min p < 0.01 for salbutamol-ipratropium vs. salbutamol and ipratropium. A total 4.7 doses of salbutamol were needed to improve the asthma attack, 5.3 of ipratropium and 3.7 of salbutamol-ipratropium, with p < 0.01 for salbutamol-ipratropium vs. salbutamol and ipratropium. We conclude that the combination of salbutamol and ipratropium is more effective than each medication alone in treating acute asthma attacks in pediatric patients.

OBJECTIVE

To investigate the changes of antisperm antibodies (AsAb), sexual hormones, and inhibin B (INH B) in patients before and after testicular torsion, as well as the effects of these factors on testicular function and reproduction.

METHODS

Ten patients with single acute testicular torsion (left side 9 and right side 1), aged 16-45 years (19.6 on average), disease course of 3-6 days (averaging 4.7 days), underwent surgical removal of the damaged testis. Before and after the operation, serum AsAb (IgG, IgM, IgA) and INH B were measured by ELISA, and serum follicle-stimulating hormone (FSH), luteotropic hormone (LH), and testosterone (T) determined by chemoluminescence autoanalyzer.

RESULTS

After the operation, the AsAb levels rose significantly and remained high for at least 26 weeks. The level of INH B was the lowest in the 3rd week and restored to normal in the 12th week, with significant difference between preoperation and the 3rd or the 6th week after the operation. The levels of LH and INH B in the 26th week were elevated significantly compared with the 6th.

CONCLUSIONS

Testicular injury induced the elevation of AsAb, which would last a very long time. The change of INH B was closely related with the injury of the testis, which reflected the degree of testicular injury and functional restoration of the patients after the operation. Our study showed that AsAb and INH B can be used as useful tools for monitoring testicular function and reproduction.

The serpin C1 inhibitor (C1-INH) is the only regulator of classical complement activation as well as the major regulator of the contact system. Its importance is demonstrated by hereditary angioedema (HAE), a severe disease with potentially life-threatening attacks due to deficiency or dysfunction of C1-INH. C1-INH replacement is the therapy of choice in HAE. In addition, C1-INH showed to have beneficial effects in other diseases characterized by inappropriate complement and contact system activation. Due to some limitations of its clinical application, there is a need for improving the efficacy of therapeutically applied C1-INH or to enhance the activity of endogenous C1-INH. Given the known potentiating effect of heparin on C1-INH, sulfated glycans (SG) may be such candidates. The aim of this study was to characterize suitable SG by evaluating structure-activity relationships. For this, more than 40 structurally distinct SG were examined for their effects on C1-INH, C1s and FXIIa. The SG turned out to potentiate the C1s inhibition by C1-INH without any direct influence on C1s. Their potentiating activity proved to depend on their degree of sulfation, molecular mass as well as glycan structure. In contrast, the SG had no effect on the FXIIa inhibition by C1-INH, but structure-dependently modulated the activity of FXIIa. Among the tested SG, β-1,3-glucan sulfates with a Mr ≤ 10 000 were identified as most promising lead candidates for the development of a glycan-based C1-INH amplifier. In conclusion, the obtained information on structural characteristics of SG favoring C1-INH potentiation represent an useful elementary basis for the development of compounds improving the potency of C1-INH in diseases and clinical situations characterized by inappropriate activation of complement and contact system.

BACKGROUND

Intraportal islet transplantation (ITx) causes instant blood-mediated inflammatory reaction (IBMIR), resulting in an early loss of transplanted islets. Porcine islets, transplanted intraportally into nonhuman primates (NHPs), induce complement activation, contributing to the development of IBMIR; however, the exact mechanism is not clear.

METHODS

Complement activation were compared after incubation of purified adult porcine islets in 20% human serum with or without complement inhibitors, C1 esterase inhibitor (C1E-inh), anti-factor B, and purified human factor H. Intraportal porcine ITx was performed in diabetic NHPs to which cobra venom factor (CVF), factor H, or none of complement inhibitor was administered during the peritransplant period. The extent of complement activation and function of islet grafts were monitored after ITx.

RESULTS

The incubation of porcine islets with human serum resulted in generation of C3a, C4d, and factor Bb in the fluid phase. However, the generation of C3a after incubation was suppressed by anti-factor B or factor H, but not by C1E-inh. Moreover, in NHPs with porcine ITx, the administration of CVF or factor H ameliorated the increase in plasma C3a and factor Bb levels, as well as early release of porcine C-peptide after ITx. Furthermore, the functional survival of islet grafts was prolonged in the recipients of the CVF group compared to those in the control group.

CONCLUSIONS

The alternative complement pathway contributes to the development of IBMIR and the early loss of grafts in NHPs with porcine ITx. Complement inhibition during the peritransplant period may be beneficial for the survival of islet grafts.

This study was designed to evaluate the detrimental effect of atrazine (ATR) on germinal epitheliums (GE) cytoplasmic carbohydrate (CH) and unsaturated fatty acids (UFA) ratio and to clarify the effect of ATR on serum levels of FSH, LH, testosterone and inhibin-B (INH-B). The impact of ATR exposure on total antioxidant capacity (TAC), sperm DNA packing and integrity were also investigated. Seventy two Wistar rats were used. The rats in control group received vehicle and the animals in test groups received 100, 200 and 300 mg kg(-1) BW of ATR orally on daily bases for 12, 24 and 48 days. In ATR-received groups the spermatogenesis cell were presented with dense reactive sites for lipidophilic staining associated with faint cytoplasmic CH accumulation. Dissociated germinal epithelium, negative tubular and repopulation indexes were manifested. The serum levels of testosterone, FSH, LH and INH-B decreased by 85% after 48 days exposure to high dose of ATR. TAC was reduced in a time- and dose-dependent manner. The sperm DNA damage was marked in animals which exposed to high dose of ATR (72.50 ± 2.25%) and the percentage of nuclear immature sperm increased up to 83.40 ± 0.89%. In conclusion, ATR not only induced its detrimental effect on the endocrine function of the testes and pituitary gland but also affected the cytoplasmic CH ratio and consequently leads to inadequate energy supplement in spermatogenesis cells. Therefore the imbalanced oxidative stress occurs in testicular tissue, which in turn enhances the sperm DNA disintegrity and nuclear immaturity.

Angioneurotic edema results from acquired or genetic deficiency of C1 inhibitor (C1 INH), a member of the serpin family of protease inhibitors. C1 INH is the only plasma protease inhibitor of activated C1r and C1s, the serine protease subcomponents of the first complement component. It is also the major inhibitor of plasma kallikrein and of coagulation factor XIIa. C1 INH consists of a single polypeptide chain of 478 amino acid residues. It is the most heavily glycosylated plasma protein; a large portion of the carbohydrate is O-linked to serine and threonine residues. Hereditary angioneurotic edema (HANE) occurs in individuals heterozygous for deficiency of C1 INH. Most patients have absolute deficiency of C1 INH (type 1 HANE), while others (15% of kindred) synthesize a dysfunctional C1 INH protein. The molecular genetic defects in the C1 INH gene in both type 1 and type 2 HANE currently are being defined. Acquired angioneurotic edema (AANE) also is of two types. One of these occurs in individuals with B-cell lymphoproliferative disorders (type 1) and the other is characterized by the presence of autoantibodies directed toward the C1 INH molecule.

We have recently demonstrated that 19-hydroxyeicosatetraenoic acid (19-HETE) is the major subterminal-HETE formed in the heart tissue, and its formation was decreased during cardiac hypertrophy. In the current study, we examined whether 19-HETE confers cardioprotection against angiotensin II (Ang II)-induced cardiac hypertrophy. The effect of Ang II, with and without 19-HETE (20 μM), on the development of cellular hypertrophy in cardiomyocyte RL-14 cells was assessed by real-time PCR. Also, cardiac hypertrophy was induced in Sprague-Dawley rats by Ang II, and the effect of increasing 19-HETE by isoniazid (INH; 200mg/kg/day) was assessed by heart weight and echocardiography. Also, alterations in cardiac cytochrome P450 (CYP) and their associated arachidonic acid (AA) metabolites were determined by real-time PCR, Western blotting and liquid-chromatography-mass-spectrometry. Our results demonstrated that 19-HETE conferred a cardioprotective effect against Ang II-induced cellular hypertrophy in vitro, as indicated by the significant reduction in β/α-myosin heavy chain ratio. In vivo, INH improved heart dimensions, and reversed the increase in heart weight to tibia length ratio caused by Ang II. We found a significant increase in cardiac 19-HETE, as well as a significant reduction in AA and its metabolite, 20-HETE. In conclusion, 19-HETE, incubated with cardiomyocytes in vitro or induced in the heart by INH in vivo, provides cardioprotection against Ang II-induced hypertrophy. This further confirms the role of CYP, and their associated AA metabolites in the development of cardiac hypertrophy.

Efficacy of preventive chemotherapy in tuberculosis-infected children depends to a great extend on medical compliance and drug tolerability. Two new short-course chemoprevention-regimes of tuberculosis--four months Rifampin (A) and two months Rifampin plus Pyrazinamide (B)--were compared with the well established regimen of six months Isoniacid (C). 150 children (mean age 3.6 years with Tb conversion) were randomly allocated to these three regimens. 13 patients were non-compliant, in terms of interview, urinary INH-test strips, urine colour and prescription frequency: 7 in group C and 3 in group A and B, respectively. Adverse effects were observed in 5 patients: 3 in group C and 1 in group A and B. 1 child (group B) developed tuberculosis two years after stopping short course chemoprevention. Good compliance (94%) as well as neglectable risks of adverse effects (2%) justify further controlled studies to evaluate the efficacy of short course chemoprevention in childhood.