Effectiveness of voice therapy in reflux-related voice disorders

Gastroesophageal reflux (GER) with laryngopharyngeal reflux plays a significant role in voice disorders. A significant proportion of patients attending ear, nose, and throat clinics with voice disorders may have gastresophageal reflux disease (GERD). There is no controlled study of the effect of voice therapy on GERD. We assessed the effect of voice therapy in patients with dysphonia and GERD. Thirty‐two patients with dysphonia and GERD underwent indirect laryngoscopy and voice analysis. Esophageal and laryngeal symptoms were assessed using the reflux symptom index (RSI). At endoscopy, esophagitis was graded according to Los Angeles classification. Patients were randomized to receive either voice therapy and omeprazole (20 mg bid) (n = 16, mean [SD] age 36.1 [9.6] y; 5 men; Gp A) or omeprazole alone (n = 16, age 31.8 [11.7] y; 9 men; Gp B). During voice analysis, jitter, shimmer, harmonic‐to‐noise ratio (HNR) and normalized noise energy (NNE) were assessed using the Dr. Speech software (version 4 1998; Tigers DRS, Inc). Hoarseness and breathiness of voice were assessed using a perceptual rating scale of 0–3. Parameters were reassessed after 6 weeks, and analyzed using parametric or nonparametric tests as applicable. In Group A, 9 patients had Grade A, 3 had Grade B, and 1 had Grade C esophagitis; 3 had normal study. In Group B, 8 patients had Grade A, 2 had Grade B esophagitis, and 6 had normal study. Baseline findings: median RSI scores were comparable (Group A 20.0 [range 14–27], Group B 19.0 [15–24]). Median rating was 2.0 for hoarseness and breathiness for both groups. Values in Groups A and B for jitter 0.5 (0.6) versus 0.5 (0.8), shimmer 3.1 (2.5) versus 2.8 (2.0), HNR 23.0 (5.6) versus 23.1 (4.2), and NNE ?7.3 (3.2) versus ?7.2 (3.4) were similar. Post‐therapy values for Groups A and B: RSI scores were 9.0 (5–13; P < 0.01 as compared with baseline) and 13.0 (10–17; P < 0.01), respectively. Ratings for hoarseness and breathiness were 0.5 (P < 0.01) and 1.0 (P < 0.01) and 2.0. Values for jitter were 0.2 (0.0; P = 0.02) versus 0.4 (0.7), shimmer 1.3 (0.7; P < 0.01) versus 2.3 (1.2), HNR 26.7 (2.3; P < 0.01) versus 23.7 (3.2), and NNE ?12.3 (3.0, P < 0.01) versus ?9.2 (3.4; P < 0.01). Improvement in the voice therapy group was significantly better than in patients who received omeprazole alone. Dysphonia is a significant problem in GER. Treatment for GER improves dysphonia, but in addition, voice therapy enhances the improvement.     

A unique C2 domain at the C terminus of Munc13 promotes synaptic vesicle priming

Neurotransmitter release during synaptic transmission comprises a tightly orchestrated sequence of molecular events, and Munc13-1 is a cornerstone of the fusion machinery. A forward genetic screen for defects in neurotransmitter release in Caenorhabditis elegans identified a mutation in the Munc13-1 ortholog UNC-13 that eliminated its unique and deeply conserved C-terminal module (referred to as HC2M) containing a Ca²⁺-insensitive C2 domain flanked by membrane-binding helices. The HC2M module could be functionally replaced in vivo by protein domains that localize to synaptic vesicles but not to the plasma membrane. HC2M is broadly conserved in other Unc13 family members and is required for efficient synaptic vesicle priming. We propose that the HC2M domain evolved as a vesicle/endosome adaptor and acquired synaptic vesicle specificity in the Unc13ABC protein family.

Chemical synaptic transmission is the primary mode of cellular communication within the nervous system. The presynaptic piece of this process encompasses a remarkable set of sequential and highly regulated interactions between a host of proteins, synaptic vesicles (SV), the plasma membrane, and calcium ions (Ca²⁺). Fusion of neurotransmitter-containing vesicles with the presynaptic plasma membrane is driven by the assembly of the neuronal SNAREs SNAP-25 and Syntaxin 1 on the plasma membrane and Synaptobrevin-2/VAMP2 on the SV. The assembly process and its coupling to intracellular Ca²⁺ are choreographed by a deeply conserved group of proteins including Munc13, Munc18, Synaptotagmin 1, and Complexin (1–4). Together with the SNAREs, these proteins form the core of the fusion apparatus across all metazoan nervous systems (5–7).First identified in a landmark genetic screen for nervous system mutants in the nematode Caenorhabditis elegans, UNC-13 is the founding member of the highly conserved metazoan Unc13 secretory protein family that includes Unc13ABC in humans (Munc13-1/2/3 in mice) (8–10). Munc13-1/UNC-13 localizes to the presynaptic active zone and is implicated in numerous presynaptic functions including initiation of release site assembly, SV docking and priming, Ca²⁺- and lipid-dependent forms of short-term synaptic plasticity, opening and positioning Syntaxin 1 for SNARE assembly, and protecting SNARE complexes from disassembly by NSF/alpha-SNAP (3, 11–13). Loss of Munc13-1 orthologs in the nervous system almost entirely eliminates all forms of chemical synaptic transmission, establishing the Unc13 family as essential to this process (14–16). All UNC-13 orthologs contain a large Syntaxin-binding MUN domain flanked by a Ca²⁺- and lipid-binding C1-C2 module and an additional C2 domain on its C terminus referred to as C2C (5, 10, 17).The C-terminal end of UNC-13 is the least understood domain within the Unc13 protein family in terms of both structure and mechanism (18, 19). Recent work on the MUN and C2C domains of Munc13-1 both in vitro and in cultured hippocampal synapses supports the notion that the MUN-C2C region attaches Munc13-1 to SVs as a means of preparing SVs for fusion (20, 21), but several questions remain unresolved. Is the SV interaction mediated by direct membrane binding? Does the C2C domain itself bind to SVs or does the MUN domain serve this role? Does either domain provide cargo specificity as part of the priming process? Interestingly, the C-terminal end of the MUN domain of CAPS, another Unc13 family member, can bind dense-core vesicles (DCVs) although it lacks a C-terminal C2 domain (22). Moreover, the MUN domain without the C2C domain has also been demonstrated to bind liposomes through an interaction with Synaptobrevin 2 (23). These observations bring up several possibilities for interactions with the C terminus of Munc13 including direct MUN–membrane interactions, C2C–membrane interactions, or protein–protein interactions involving either or both domains. Other Unc13 family members possessing a MUN domain with a C-terminal C2 domain such as Unc13D/Munc13-4 and BAIAP3 have been proposed to tether specific cargo such as endosomes, secretory granules, and large DCVs (24, 25). How Unc13 proteins select among different cargos remains largely unanswered (24, 26, 27).Through behavioral, electrophysiological, biochemical, and genetic approaches, we uncover a deeply conserved C-terminal membrane-binding domain within Munc13-1/UNC-13 termed the Munc13 C-terminal (MCT) domain. This region, together with C2C and a neighboring N-terminal helix fold together into a stable membrane-binding protein domain in vitro, and loss of any part of this module in vivo impairs SV priming and nervous system function. Moreover, the C-terminal domain can be replaced by foreign domains that bind SVs but not the plasma membrane, demonstrating a role in SV interactions at the synapse. Phylogenetic protein sequence comparisons suggest that the ancestral Unc13/BAIAP3 homolog possessed a similar C-terminal domain prior to the emergence of metazoa, and subsequently, the UNC-13ABC subfamily domain evolved as an SV adaptor that plays a critical role in neurotransmission in all animals.     

Studies of Antiviral Activity and Cytotoxicity of Wrightia tinctoria and Morinda citrifolia

Different extracts of leaf parts of Wrightia tinctoria and fruit powder of Morinda citrifolia have been studied against replication of HIV-1(IIIB) in MT-4 cells and HCV in Huh 5.2 cells. Chloroform extract of Wrightia tinctoria exhibited a maximum protection of 48% against the cytopathic effect of HIV-1(IIIB) in MT-4 cells. Fruit juice of Morinda citrifolia exhibited a displayed marked cytotoxic activity in lymphocyte (MT-4) cells (CC50: 0.19 mg/ml). The 50% effective concentration for inhibition of HCV subgenomic replicon replication in Huh 5-2 cells by Morinda citrifolia was 0.98 μg/ml and by chloroform extract of Wrightia tinctoria was 10 μg/ml. The concentration that reduced the growth of exponentially proliferating Huh 5-2 cells by 50% was greater than 50 μg/ml.     

Synthesis, Antiviral and Cytotoxic Activities of 2-(2-Phenyl carboxylic acid)-3-Phenylquinazolin -4(3H)-one Derivatives

A series of novel 2,3-disubstitutedquinazolin-4(3H)-ones have been synthesized by condensation of 2-substituted benzo[1,3]oxazine-4-ones and anthranilic acid. Synthesized compounds were evaluated for in vitro antiviral activity against HIV, HSV and vaccinia viruses. 5-Bromo-2-(6-bromo-4-oxo-2-phenyl-4H-quinazolin-3-yl)-benzoic acid (MBR2) exhibited distinct antiviral activity against Herpes simplex and vaccinia viruses.     

Antitumour Activity of Bauhinia variegata Against Ehrlich Ascites Carcinoma Induced Mice

The antitumour activity of ethanol extract of Bauhinia variegata (EBV) has been evaluated against Ehrlich ascites carcinoma (EAC) in Swiss albino mice. A significant enhancement of mean survival time of EBV treated tumour bearing mice was found with respect to the control group. EBV treatment was found to enhance peritoneal cell counts. After 14 days of inoculation, EBV is able to reverse the changes in the haemotological parameters, protein and PCV consequent to tumour inoculation. Oral administration of EBV was effective in reducing solid tumour mass development induced by EAC cells. EBV was found to be a potent cytotoxic towards EAC tumour cells.     

LC-MS/MS-ESI method for simultaneous quantitation of three endocannabinoids and its application to rat pharmacokinetic studies

An LC-MS/MS-ESI method has been validated for simultaneous estimation of the three endocannabinoids; N-arachidonoylethanolamine (AEA), N-oleoylethanolamine (OEA) and palmitoylethanolamide (PEA), in surrogate matrix using AEA-d (4) as an internal standard with highest sensitivity over the existing methods. Simple precipitation was used to extract analytes and these were subsequently analyzed on a monolithic column. Linear response function was established over the concentration range 12.3 to 1225 pg/ml for AEA (r > 0.994); 0.70 to 641 ng/ml for OEA (r > 0.999) and 0.54 to 321 ng/ml (r > 0.998) for PEA. The intra- and inter-day precision values met the acceptance to criteria as per US FDA guidelines. Analytes were found to be stable in the battery of stability studies. The method was applied to quantify endogenous levels of analytes in rat plasma.     

A glance over the fence: Using phylogeny and species comparison for a better understanding of antigen recognition by human γδ T-cells

Both, jawless and jawed vertebrates possess three lymphocyte lineages defined by highly diverse antigen receptors: Two T-cell- and one B-cell-like lineage. In both phylogenetic groups, the theoretically possible number of individual antigen receptor specificities can even outnumber that of lymphocytes of a whole organism. Despite fundamental differences in structure and genetics of these antigen receptors, convergent evolution led to functional similarities between the lineages. Jawed vertebrates possess αβ and γδ T-cells defined by eponymous αβ and γδ T-cell antigen receptors (TCRs). “Conventional” αβ T-cells recognize complexes of Major Histocompatibility Complex (MHC) class I and II molecules and peptides. Non-conventional T-cells, which can be αβ or γδ T-cells, recognize a large variety of ligands and differ strongly in phenotype and function between species and within an organism. This review describes similarities and differences of non-conventional T-cells of various species and discusses ligands and functions of their TCRs. A special focus is laid on Vγ9Vδ2 T-cells whose TCRs act as sensors for phosphorylated isoprenoid metabolites, so-called phosphoantigens (PAg), associated with microbial infections or altered host metabolism in cancer or after drug treatment. We discuss the role of butyrophilin (BTN)3A and BTN2A1 in PAg-sensing and how species comparison can help in a better understanding of this human Vγ9Vδ2 T-cell subset.     

Long‐term administration of AMD3100, an antagonist of SDF‐1/CXCR4 signaling,alters fracture repair

Fracture healing involves rapid stem and progenitor cell migration, homing, and differentiation. SDF‐1 (CXCL12) is considered a master regulator of CXCR4‐positive stem and progenitor cell trafficking to sites of ischemic (hypoxic) injury and regulates their subsequent differentiation into mature reparative cells. In this study, we investigated the role of SDF‐1/CXCR4 signaling in fracture healing where vascular disruption results in hypoxia and SDF‐1 expression. Mice were injected with AMD3100, a CXCR4 antagonist, or vehicle twice daily until euthanasia with the intent to impair stem cell homing to the fracture site and/or their differentiation. Fracture healing was evaluated using micro‐computed tomography, histology, quantitative PCR, and mechanical testing. AMD3100 administration resulted in a significantly reduced hyaline cartilage volume (day 14), callus volume (day 42) and mineralized bone volume (day 42) and reduced expression of genes associated with endochondral ossification including collagen Type 1 alpha 1, collagen Type 2 alpha 1, vascular endothelial growth factor, Annexin A5, nitric oxide synthase 2, and mechanistic target of rapamycin. Our data suggest that the SDF‐1/CXCR4 signaling plays a central role in bone healing possibly by regulating the recruitment and/or differentiation of stem and progenitor cells. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:1853–1859, 2012     

Spontaneous esophageal perforation in a patient with achalasia cardia and rheumatoid arthritis.

Perforation of stasis ulcers in achalasia cardia has not been reported in literature. We report a 45-year-old lady with achalasia and rheumatoid arthritis who developed perforation and esophago-mediastinal sinus at the site of stasis ulcers. She succumbed to respiratory infection after resection of the sinus tract, Heller's cardiomyotomy, cervical esophagostomy and feeding jejunostomy.