Developmental regulation of beta-carboline-induced inhibition of glycine-evoked responses depends on glycine receptor beta subunit expression

In this work, we show that beta-carbolines, which are known negative allosteric modulators of GABA(A) receptors, inhibit glycine-induced currents of embryonic mouse spinal cord and hippocampal neurons. In both cell types, beta-carboline-induced inhibition of glycine receptor (GlyR)-mediated responses decreases with time in culture. Single-channel recordings show that the major conductance levels of GlyR unitary currents shifts from high levels (> or = 50 pS) in 2 to 3 days in vitro (DIV) neurons to low levels (<50 pS) in 11 to 14 DIV neurons, assessing the replacement of functional homomeric GlyR by heteromeric GlyR. In cultured spinal cord neurons, the disappearance of beta-carboline inhibition of glycine responses and high conductance levels is almost complete in mature neurons, whereas a weaker decrease in beta-carboline-evoked glycine response inhibition and high conductance level proportion is observed in hippocampal neurons. To confirm the hypothesis that the decreased sensitivity of GlyR to beta-carbolines depends on beta subunit expression, Chinese hamster ovary cells were permanently transfected either with GlyR alpha2 subunit alone or in combination with GlyR beta subunit. Single-channel recordings revealed that the major conductance levels shifted from high levels (> or = 50 pS) in GlyR-alpha2-transfected cells to low levels (<50 pS) in GlyR-alpha2+beta-containing cells. Consistently, both picrotoxin- and beta-carboline-induced inhibition of glycine-gated currents were significantly decreased in GlyR-alpha2+beta-transfected cells compared with GlyR-alpha2-containing cells. In summary, we demonstrate that the incorporation of beta subunits in GlyRs confers resistance not only to picrotoxin but also to beta-carboline-induced inhibition. Furthermore, we also provide evidence that hippocampal neurons undergo in vitro a partial maturation process of their GlyR-mediated responses.     

Striatal PSA-NCAM(+) precursor cells from the newborn rat express functional glycine receptors

Immunocytochemical analysis showed that ionotropic glycine receptors are expressed in neurogenic progenitors purified from the newborn rat striatum and expressing the polysialylated form of the neural cell adhesion molecule, both in vitro and in situ. To ascertain whether glycine receptors were functional in vitro, whole-cell patch-clamp recordings demonstrated that glycine triggers inward strychnine-sensitive currents in the majority of these cells. Moreover, we found that glycine receptors expressed by these neurogenic progenitors display intermediate electrophysiological characteristics between those of glycine receptors expressed by neural stem cells and by mature interneurons from the rat striatum. Altogether, the present data show that functional strychnine-sensitive glycine receptors are expressed in neurogenic progenitors purified from the newborn rat striatum.     

Iatrogenic bile duct strictures: a review of 22 cases

The incidence of iatrogenic bile duct strictures in Ethiopia appears to be increasing. Of 27 patients that sustained bile duct injuries at open cholecystectomy, admitted during May 1996 to December 2002, 22 cases of bile duct strictures are presented to evaluate outcome of treatment. The mean age was 40 years, 15 females. Twenty-one were referrals. The usual presenting features were biliary peritonitis and jaundice. The average time lapse between the original surgery and admission to hospital was eight months. About 73% had Bismuth grade III-IV strictures and all patients underwent Roux-en-Y hepatico-jejunostomy. Postoperatively, biliary-cutaneous fistula, recurrent ascending cholangitis and wound infection were observed frequently. The overall mortality rate was 13.6%. Bile duct injuries and strictures occur in young productive age groups. Prevention of the occurrence of bile duct injury and its progression to a devastating stricture reduces morbidity and mortality.     

Down-regulation of the IGF-2/H19 locus during normal and malignant hematopoiesis is independent of the imprinting pattern

H19 and IGF-2 are two growth regulatory genes located on chromosome 11p15 implicated in tumorigenesis. Both genes are imprinted and regulated reciprocally under many circumstances. In order to elucidate the contribution of H19 and IGF-2 to leukemogenesis, the mRNA expression level of both genes were quantitated in bone marrow biopsies and peripheral blood samples from normal (n=98), chronic myelomonocytic leukemia (CMML, n=43), chronic myelogenous leukemia (CML, n=40) and, acute myelogenous leukemia (AML, n=32) cases. A concomitant reduction of H19 and IGF-2 expression was observed in all leukemic samples compared to the healthy controls. This down-regulation was not accompanied by changes in methylation of the differentially methylated region (DMR). Whereas the H19 gene showed strict monoallelic expression in all informative normal (n=31) and leukemic (n=54) samples, the imprinting pattern of the IGF2 gene was found to be heterogeneous. No correlations between imprinting status (mono- versus biallelic expression), quantitative mRNA expression levels and course of disease were found for the IGF-2 gene. The data suggest a disturbed regulation of the IGF-2/H19 locus in myeloid leukemias which is not caused by loss of imprinting.     

Modified Subtotal Lothrop Procedure for Extended Frontal Sinus and Anterior Skull Base Access: A Cadaveric Feasibility Study with Clinical Correlates

Objective The endoscopic modified Lothrop procedure (EMLP) is an established approach for recalcitrant frontal sinus disease and anterior skull base exposure. However, in select cases, this technique may involve unnecessary resection of sinonasal structures. In this study, we propose a modification of the EMLP, termed the modified subtotal-Lothrop procedure (MSLP), to access the anterior skull base and complex frontal sinus disease for which access to the bilateral frontal sinus posterior table is required.Methods A cadaveric dissection with photo documentation was performed at an academic medical center on four cadaver heads using standard endoscopic techniques to demonstrate the MSLP and its feasibility.Results The endoscopic MSLP allowed ample access for instrumentation in each of the dissections using a 30- or 70-degree endoscope. Adequate bilateral access to the posterior table of the frontal sinus was gained in all cases without the need for dissection of the contralateral frontal sinus recess (FSR).Conclusion The MSLP appears to be a feasible technique for exposure of the anterior skull base and accessing complex frontal sinus pathology. This modification provides similar anterior skull base exposure and surgical maneuverability as the EMLP while limiting surgical dissection to one FSR, thereby preserving as much of the natural mucociliary drainage pathways as possible.     

Factors associated with complete immunization coverage in children aged 12¿23 months in Ambo Woreda, Central Ethiopia

ABSTRACT: BACKGROUND: Vaccination is a proven tool in preventing and eradicating communicable diseases, but a considerable proportion of childhood morbidity and mortality in Ethiopia is due to vaccine preventable diseases. Immunization coverage in many parts of the country remains low despite the efforts to improve the services. In 2005, only 20% of the children were fully vaccinated and about 1 million children were unvaccinated in 2007. The objective of this study was to assess complete immunization coverage and its associated factors among children aged 12-23 months in Ambo woreda. METHODS: A cross-sectional community-based study was conducted in 8 rural and 2 urban kebeles during January- February, 2011. A modified WHO EPI cluster sampling method was used for sample selection. Data on 536 children aged 12-23 months from 536 representative households were collected using trained nurses. The data collectors assessed the vaccination status of the children based on vaccination cards or mother's verbal reports using a pre-tested structured questionnaire through house-to-house visits. Bivariate and multivariate logistic regression analyses were used to assess factors associated with immunization coverage. RESULTS: About 96% of the mothers heard about vaccination and vaccine preventable diseases and 79.5% knew the benefit of immunization. About 36% of children aged 12-23 months were fully vaccinated by card plus recall, but only 27.7% were fully vaccinated by card alone and 23.7% children were unvaccinated. Using multivariate logistic regression models, factors significantly associated with complete immunization were antenatal care follow-up (adjusted odds ratio(AOR=2.4, 95% CI: 1.2- 4.9), being born in the health facility (AOR=2.1, 95% CI: 1.3-3.4), mothers' knowledge about the age at which vaccination begins (AOR= 2.9, 95% CI: 1.9-4.6) and knowledge about the age at which vaccination completes (AOR=4.3, 95% CI: 2.3-8), whereas area of residence and mother's socio-demographic characteristics were not significantly associated with full immunization among children. CONCLUSION: Complete immunization coverage among children aged 12-23 months remains low. Maternal health care utilization and knowledge of mothers about the age at which child begins and finishes vaccination are the main factors associated with complete immunization coverage. It is necessary that, local interventions should be strengthened to raising awareness of the community on the importance of immunization, antenatal care and institutional delivery.     

Equine Herpesvirus‐1 Myeloencephalopathy,an Emerging Threat of Working Equids in Ethiopia

Although equine herpesvirus myeloencephalopathy (EHM) is a sporadic and relatively uncommon manifestation of equine herpesvirus‐1 (EHV‐1), it has the potential for causing devastating outbreaks in horses. Up till now, there were no reported EHM outbreaks in donkeys and mules. This study describes the isolation and molecular characterization of EHV‐1 from clinically EHM‐affected horses (n = 6), mules (n = 3) and donkeys (n = 82) in Ethiopia during outbreaks from May 2011 to December 2013. The incidence of EHM cases was higher from April to mid‐June. EHM in donkeys was more severe and death without clinical signs of paralysis, and recumbency was frequently observed. The main age of affected equines ranged from 7 to 10 years (n = 51; 56.0%), and females (n = 58; 63.7%) were more affected than males. The incidence of neuropathogenic (D₇₅₂) and non‐neuropathogenic (N₇₅₂) variants of EHV‐1 from EHM‐affected equines in Ethiopia was assessed by sequencing the DNA polymerase gene (ORF30) of the EHV‐1 isolates. The results indicated that from the total of 91 clinically affected equines, 90 (98.9%) of them had an ORF30 D₇₅₂ genotype. An ORF30 N₇₅₂ variant was only found in one donkey. Analysis of ORF68 as grouping marker for geographical differences showed that the Ethiopian EHV‐1 isolates belong to geographical group 4. Due to the fatal nature of EHV‐1 in donkeys, it would be interesting to examine the pathogenesis of EHM in this species. At present, there is no vaccine available in Ethiopia, and therefore, outbreaks of EHV‐1 should be controlled by proper management adaptations. In addition, it is important to test the efficacy of the commercial vaccines not only in horses, but also in donkeys and mules.     

Magnetite nanoparticle decorated reduced graphene oxide for adsorptive removal of crystal violet and antifungal activities

This work reports the synthesis and application of magnetic rGO/Fe₃O₄ NCs using a pod extract of Dolichos lablab L. as areducing agent. GO was synthesized by a modified Hummers method, however GO was reduced using the plant extract to produce rGO. The as-synthesized rGO/Fe₃O₄ NCs were characterized by UV-vis spectrophotometer, Fourier transform infrared (FT-IR) spectroscopy, FT-Raman spectroscopy, X-ray diffraction (XRD), field emission scanning electron microscopy supported with energy dispersed X-ray spectroscopy (FESEM-EDX), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM). The synthesis of magnetic rGO/Fe₃O₄ NCs was confirmed from characterization results of FT-Raman, TEM and VSM. The FT-Raman results showed the D and G bands at 1306.92 cm⁻¹ and 1591 cm⁻¹ due to rGO and a peak at around 589 cm⁻¹ due to Fe₃O₄ NPs that were anchored on rGO sheets; TEM results showed the synthesis of Fe₃O₄ with an average particle size of 8.86 nm anchored on the surface of rGO sheets. The VSM result confirmed the superparamagnetic properties of the rGO/Fe₃O₄ NCs with a saturation magnetization of 42 emu g⁻¹. The adsorption capacity of rGO/Fe₃O₄ NCs towards crystal violet (CV) dye was calculated to be 62 mg g⁻¹. The dye removal behavior fitted well with the Freundlich isotherm and the pseudo-second-order kinetic model implies possible chemisorption. Besides, rGO/Fe₃O₄ NCs showed antifungal activities against Trichophyton mentagrophytes and Candida albicans by agar-well diffusion method with a zone inhibition of 24 mm and 21 mm, respectively. Therefore, rGO/Fe₃O₄ NCs can be used as an excellent adsorbent to remove organic dye pollutants and kill pathogens.

This work reports synthesis of magnetic rGO/Fe₃O₄ NCs using pod extract of Dolichos lablab L. as a reducing agent and its applications.