Post-operative hearing among patients with labyrinthine fistula as a complication of cholesteatoma using "under water technique".

INTRODUCTION: During surgery in patients with labyrinthine fistula the mandatory complete removal of the cholesteatoma while preserving inner ear and vestibular function is a challenge. Options so far have been either the complete removal of the cholesteatoma or leaving the matrix on the fistula. We evaluated an alternative "under water" surgical technique for complete cholesteatoma resection, in terms of preservation of postoperative inner ear and vestibular function. METHODS: From 2013 to 2019, 20 patients with labyrinthine fistula due to cholesteatoma were operated. We used the canal wall down approach and removal of matrix on the fistula was done as the last step during surgery using the "under water technique". The pre and postoperative hearing tests and the vestibular function were retrospectively examined. RESULTS: There was no significant difference between pre and post-operative bone conduction thresholds; 20% experienced an improvement of more than 10 dB, with none experiencing a postoperative worsening of sensorineural hearing loss. Among seven patients who presented with vertigo, two had transient vertigo postoperatively but eventually recovered. CONCLUSION: Our data show that the "under water technique" for cholesteatoma removal at the labyrinthine fistula is a viable option in the preservation of inner ear function and facilitating complete cholesteatoma removal.

Nasal self-packing for epistaxis in Hereditary Hemorrhagic Telangiectasia increases quality of life.

STATEMENT OF PROBLEM: Hereditary hemorrhagic telangiectasia (HHT) is characterized by recurrent epistaxis that can lead to a feeling of losing control. We assessed potential benefits and side effects of different nasal packings used by patients themselves. METHOD OF STUDY: An online-questionnaire in English and German was used to analyze nasal self-packings. RESULTS: 588 of the 915 respondents suffered from HHT with most of them having moderate or severe epistaxis. Almost two thirds of the patients had already treated themselves with nasal packings. While one quarter used non-pneumatic nasal packings (NPNP) or pneumatic nasal packings (PNP), nearly half of the patients took only tissues to stop the bleeding. Patients with severe epistaxis used PNP more often than NPNP. Using nasal packings, most patients could stop their nosebleeds after a while. Patients using PNP reported the feeling of losing control less often and significant improvements in quality of life with a positive GBI. CONCLUSIONS: Our study showed that most patients with HHT using nasal self-packings could stop the bleeding after a while. Nasal self-packing is a user-friendly and secure method leaving patients more self-confident and independent.

Structural basis for the allosteric inhibitory mechanism of human kidney-type glutaminase (KGA) and its regulation by Raf-Mek-Erk signaling in cancer cell metabolism.

Besides thriving on altered glucose metabolism, cancer cells undergo glutaminolysis to meet their energy demands. As the first enzyme in catalyzing glutaminolysis, human kidney-type glutaminase isoform (KGA) is becoming an attractive target for small molecules such as BPTES [bis-2-(5 phenylacetamido-1, 2, 4-thiadiazol-2-yl) ethyl sulfide], although the regulatory mechanism of KGA remains unknown. On the basis of crystal structures, we reveal that BPTES binds to an allosteric pocket at the dimer interface of KGA, triggering a dramatic conformational change of the key loop (Glu312-Pro329) near the catalytic site and rendering it inactive. The binding mode of BPTES on the hydrophobic pocket explains its specificity to KGA. Interestingly, KGA activity in cells is stimulated by EGF, and KGA associates with all three kinase components of the Raf-1/Mek2/Erk signaling module. However, the enhanced activity is abrogated by kinase-dead, dominant negative mutants of Raf-1 (Raf-1-K375M) and Mek2 (Mek2-K101A), protein phosphatase PP2A, and Mek-inhibitor U0126, indicative of phosphorylation-dependent regulation. Furthermore, treating cells that coexpressed Mek2-K101A and KGA with suboptimal level of BPTES leads to synergistic inhibition on cell proliferation. Consequently, mutating the crucial hydrophobic residues at this key loop abrogates KGA activity and cell proliferation, despite the binding of constitutive active Mek2-S222/226D. These studies therefore offer insights into (i) allosteric inhibition of KGA by BPTES, revealing the dynamic nature of KGA's active and inhibitory sites, and (ii) cross-talk and regulation of KGA activities by EGF-mediated Raf-Mek-Erk signaling. These findings will help in the design of better inhibitors and strategies for the treatment of cancers addicted with glutamine metabolism.

Novel fluorescent protein from Hydnophora rigida possesses green emission.

Fluorescent proteins are a family of proteins capable of producing fluorescence at various specific wavelengths of ultra violet light. We have previously reported the identification and characterization of a novel cyan fluorescent protein (HriCFP) from a reef coral species, Hydnophora rigida. In search of new members of the diverse family of fluorescent proteins, here we report a new green fluorescent protein (HriGFP) from H. rigida. HriGFP was identified, cloned, expressed in Escherichia coli and purified to homogeneity by metal affinity and size exclusion chromatography. The dynamic light scattering and gel filtration experiments suggested the presence of monomers in solution. The peptide mass fingerprint on the purified protein established the identity of HriGFP. HriGFP had excitation peak at 507 nm and emission peak at 527 nm. HriGFP was similar to HriCFP except the last 16 amino acid sequence at the C-terminal; however, they have shown least similarity with other known fluorescent proteins. Moreover the computational model suggests that HriGFP is a globular protein which consists of 6 α-helices and 3 ß-sheets. Taken together our results suggested that HriGFP is a novel naturally occurring fluorescent protein that exists as a monomer in solution.

Efficacy, safety and tolerability of topical terbinafine nail solution in patients with mild-to-moderate toenail onychomycosis: results from three randomized studies using double-blind vehicle-controlled and open-label active-controlled designs.

BACKGROUND: Terbinafine nail solution (TNS) was developed for the treatment of onychomycosis. OBJECTIVE: To assess the efficacy of TNS vs. vehicle and amorolfine 5% nail lacquer. METHODS: Subjects with mild-to-moderate toe onychomycosis (25% to ≤75% nail-involvement, matrix uninvolved) were randomized to receive either TNS or vehicle in two double-blind studies, and to TNS or amorolfine in an active-controlled, open-label study. Primary endpoint was complete cure (no residual clinical involvement and negative mycology) at week 52. Secondary endpoints were mycological cure (negative mycology defined as negative KOH microscopy and negative culture) and clinical effectiveness (≤10% residual-involvement and negative mycology) at week 52. RESULTS: Complete cure was not different between TNS vs. vehicle and amorolfine. Mycological cure was higher with TNS vs. vehicle, as was clinical effectiveness with TNS vs. vehicle, and TNS and amorolfine were not different for secondary efficacy endpoints. Patients achieving mycological cure had a better clinical outcome, and efficacy was improved in subjects with milder disease. Post hoc analysis suggests that nail thickness is an important prognostic factor. Moreover, mycological cure may require 6 months of treatment regimen while complete cure and clinical effectiveness may be achievable only after 10 months. A simulation study suggests that longer treatment duration would have resulted in higher complete cure with TNS vs. vehicle. Study treatments were well-tolerated. CONCLUSION: Primary efficacy objectives were not met in the studies reported herein. Possible reasons for failure to achieve significant outcomes include insufficient length of treatment; stringency of primary endpoint and severity of nail involvement of study population.

An analysis of health-related quality of life in the phase III PROSELICA and FIRSTANA studies assessing cabazitaxel in patients with metastatic castration-resistant prostate cancer.

BACKGROUND: Men with metastatic castration-resistant prostate cancer (mCRPC) are living longer, therefore optimizing health-related quality of life (HRQL), as well as survival outcomes, is important for optimal patient care. The aim of this study was to assess the HRQL in patients with mCRPC receiving docetaxel or cabazitaxel. PATIENTS AND METHODS: PROSELICA (NCT01308580) assessed the non-inferiority of cabazitaxel 20 mg/m2 (C20) versus 25 mg/m2 (C25) in patients with mCRPC after docetaxel. FIRSTANA (NCT01308567) assessed the superiority of C25 or C20 versus docetaxel 75 mg/m2 (D75) in patients with chemotherapy-naive mCRPC. HRQL and pain were analyzed using protocol-defined, prospectively collected, Functional Assessment of Cancer Therapy-Prostate (FACT-P) and McGill-Melzack questionnaires. Analyses included definitive improvements in HRQL, maintained or improved HRQL, and HRQL over time. RESULTS: In total, 2131 patients were evaluable for HRQL across the two studies. In PROSELICA, 38.8% and 40.5% of patients receiving C20 and C25, respectively, had definitive FACT-P total score (TS) improvements. In FIRSTANA, 43.4%, 49.7%, and 44.9% of patients receiving D75, C20, and C25, respectively, had definitive FACT-P TS improvements. In both trials, definitive improvements started after cycle 1 and were maintained for the majority of subsequent treatment cycles. More than two-thirds of patients maintained or improved their FACT-P TS. CONCLUSIONS: In PROSELICA and FIRSTANA, >40% of the 2131 evaluable patients with mCRPC had definitive FACT-P TS improvements; improvements occurred early and were maintained. More than 75% of patients maintained or improved their FACT-P TS.

Genetic diversity and relationships in mulberry (genus Morus) as revealed by RAPD and ISSR marker assays.

BACKGROUND: The genus Morus, known as mulberry, is a dioecious and cross-pollinating plant that is the sole food for the domesticated silkworm, Bombyx mori. Traditional methods using morphological traits for classification are largely unsuccessful in establishing the diversity and relationships among different mulberry species because of environmental influence on traits of interest. As a more robust alternative, PCR based marker assays including RAPD and ISSR were employed to study the genetic diversity and interrelationships among twelve domesticated and three wild mulberry species. RESULTS: RAPD analysis using 19 random primers generated 128 discrete markers ranging from 500-3000 bp in size. One-hundred-nineteen of these were polymorphic (92%), with an average of 6.26 markers per primer. Among these were a few putative species-specific amplification products which could be useful for germplasm classification and introgression studies. The ISSR analysis employed six anchored primers, 4 of which generated 93 polymorphic markers with an average of 23.25 markers per primer. Cluster analysis of RAPD and ISSR data using the WINBOOT package to calculate the Dice coefficient resulted into two clusters, one comprising polyploid wild species and the other with domesticated (mostly diploid) species. CONCLUSION: These results suggest that RAPD and ISSR markers are useful for mulberry genetic diversity analysis and germplasm characterization, and that putative species-specific markers may be obtained which can be converted to SCARs after further studies.

Structural basis for the active site inhibition mechanism of human kidney-type glutaminase (KGA).

Glutaminase is a metabolic enzyme responsible for glutaminolysis, a process harnessed by cancer cells to feed their accelerated growth and proliferation. Among the glutaminase isoforms, human kidney-type glutaminase (KGA) is often upregulated in cancer and is thus touted as an attractive drug target. Here we report the active site inhibition mechanism of KGA through the crystal structure of the catalytic domain of KGA (cKGA) in complex with 6-diazo-5-oxo-L-norleucine (DON), a substrate analogue of glutamine. DON covalently binds with the active site Ser286 and interacts with residues such as Tyr249, Asn335, Glu381, Asn388, Tyr414, Tyr466 and Val484. The nucleophilic attack of Ser286 sidechain on DON releases the diazo group (N2) from the inhibitor and results in the formation of an enzyme-inhibitor complex. Mutational studies confirmed the key role of these residues in the activity of KGA. This study will be important in the development of KGA active site inhibitors for therapeutic interventions.

Evaluation of the lateral orbital approach in management of zygomatic bone fractures.

Zygomatic maxillary fractures, also known as tripod fractures, are usually the result of a direct blow to the body of the zygoma. Tripod fracture consists of (a) zygomatic arch fracture, (b) fracture of the lateral orbital wall, and (c) fracture of the inferior orbital floor. The purpose of this study is to evaluate the functional and esthetic outcome following this lateral orbital approach in the management of zygoma fracture. This study was carried out in VMS Dental College, Salem, and in a private hospital. This study was based on the experience gained from a retrospective study of the 30 lateral orbital approaches that were used in 30 patients with fractures of the zygomatic complex, which were conducted for a period of 8 years between January 2003 and January 2011. In the retrospective study, all the 30 patients were able to open the mouth completely; eyeball movements were normal; esthetically, all patients appeared normal. There were no sinusitis or visual problems in any of the studied patients. We conclude that the lateral orbital approach is an ideal option in reduction and treatment of zygomatic bone and arch fractures.

Inferior alveolar nerve block: Alternative technique.

BACKGROUND: Inferior alveolar nerve block (IANB) is a technique of dental anesthesia, used to produce anesthesia of the mandibular teeth, gingivae of the mandible and lower lip. The conventional IANB is the most commonly used the nerve block technique for achieving local anesthesia for mandibular surgical procedures. In certain cases, however, this nerve block fails, even when performed by the most experienced clinician. Therefore, it would be advantageous to find an alternative simple technique. AIM AND OBJECTIVE: The objective of this study is to find an alternative inferior alveolar nerve block that has a higher success rate than other routine techniques. To this purpose, a simple painless inferior alveolar nerve block was designed to anesthetize the inferior alveolar nerve. MATERIALS AND METHODS: This study was conducted in Oral surgery department of Vinayaka Mission's dental college Salem from May 2009 to May 2011. Five hundred patients between the age of 20 years and 65 years who required extraction of teeth in mandible were included in the study. Out of 500 patients 270 were males and 230 were females. The effectiveness of the IANB was evaluated by using a sharp dental explorer in the regions innervated by the inferior alveolar, lingual, and buccal nerves after 3, 5, and 7 min, respectively. CONCLUSION: This study concludes that inferior alveolar nerve block is an appropriate alternative nerve block to anesthetize inferior alveolar nerve due to its several advantages.

Simple and safe posterior superior alveolar nerve block.

BACKGROUND: The posterior superior alveolar nerve (PSAN) block is a dental nerve block used for profound anesthesia of the maxillary molars. Although it is being written in texts as a commonly used technique, but in dentistry it is rarely followed due to its nonreliable landmarks, variation in depth of insertion and frequent complications. The aim and objective are to find a technically simple method of the PSAN block without any complications. STUDY AND DESIGN: This study was based on the experience gained from 200 patients of 125 males and 75 female in age group of 20 to 65 years in University of Vinayaka and department of oral and maxillofacial surgery of VMS Dental College and hospital, Salem, Tamil Nadu. RESULTS: In 200 patients' positive anesthesia obtained within a period of 5 to 10 min. No visual complications reported in this study. There was no pain during and after extraction. CONCLUSION: This study shows this PSA nerve block using curved needle would avoid all complications reported in the literature. Therefore, the technique described in this study is an ideal option to anesthetize PSA nerve.

Significance of localization of mandibular foramen in an inferior alveolar nerve block.

BACKGROUND: The mandibular foramen (MF) is an opening on the internal surface of the ramus for divisions of the mandibular vessels and nerve to pass. The aim of this study is to determine the position of the MF from various anatomical landmarks in several dry adult mandibles. MATERIALS AND METHODS: A total of 102 human dry mandibles were examined, of which 93 were of dentulous and 9 were of edentulous. The measurements were taken from the anterior border of the ramus (coronoid notch) to the midportion of the MF and then from the midportion of the MF to the other landmarks such as internal oblique ridge, inferior border, sigmoid notch, and condyle were measured and recorded. RESULTS: The data were compared using Student's t-test. The MF is positioned at a mean distance of 19 mm (with SD 2.34) from coronoid notch of the anterior border of the ramus. Superio-inferiorly from the condyle to the inferior border MF is situated 5 mm inferior to the midpoint of condyle to the inferior border distance (ramus height). CONCLUSION: We conclude that failures in the anesthesia of the inferior alveolar nerve are due to the operator error and not due to the anatomical variation.

Maxillary nerve block in management of maxillary bone fractures: Our experience.

BACKGROUND AND OBJECTIVES: The objective of this study is to evaluate the intraoral high tuberocity maxillary nerve block technique in zygoma and arch fracture reduction and fixation. STUDY AND DESIGN: This study was carried out at Arvind Multi-Specialty Hospital, Namakkal, Tamil Nadu on seven male patients with zygomatic bone and arch fracture. MATERIALS AND METHODS: Intraoral high tuberocity maxillary nerve block administered in seven patients for management of isolated zygomatic bone and arch fracture. Lidocaine 2% measuring 4 mL with 1:80000 adrenaline anesthetic solutions was used to anesthetize maxillary nerve through a 3.2 cm length and 24G, needle. The following parameters were evaluated namely onset of anesthesia, nerve block duration, outcome of treatment and Patient's comfort. RESULTS: The blocks were effective and patients were comfortable without pain during initial stage of surgery, but in latter stages two patients had mild to moderate pain. Duration of block varied from 60 to 90 min while onset varied from 3 to 10 min. There were vascular punctures in three patients, however, without hematoma. CONCLUSIONS: The maxillary nerve block is a good alternative option in selective cases of zygomatic bone fracture reduction.

Genetic variability and genetic structure of wild and semi-domestic populations of tasar silkworm (Antheraea mylitta ) ecorace Daba as revealed through ISSR markers.

The genetic diversity in the wild and semi-domestic populations of Daba ecorace of Antheraea mylitta was studied to ascertain the distribution of variability within and among populations of semi-domestic bivoltine (DB), trivoltine (DT) and nature grown wild populations (DN) with inter-simple sequence repeat (ISSR) markers. A total of 138 markers were produced among 56 individuals of the three populations, of which 98% were polymorphic. For the individual populations, the percentage polymorphism was 58.69, 52.9 and 77.54 for DB, DT and DN, respectively. Average number of observed (1.791+/- 0.408) and effective alleles (1.389+/-0.348) was also high in the wild populations in comparison to the bivoltine and trivoltine semi-domestic populations. Genetic diversity (H(t)) in DB, DT and DN was 0.180+/- 0.033, 0.153+/- 0.032 and 0.235+/- 0.033, respectively and within-population genetic diversity (H(s)) ranged from 0.166 to 0.259 with a mean of 0.189. Mean gene differentiation (G(ST)) was found to be 0.25. Shanon's diversity index was 0.278, 0.237 and 0.361 for DB, DT and DN and overall it was 0.391. Gene flow (N(m)) among the populations was 1.509. The dendrogram produced by UPGMA with Dice's genetic distance matrices resulted in the formation of three major clusters separating the three populations. Considerable intra- and inter-population variability is found in all three populations. The population structure analysis further suggests that the semi-domestic populations of Daba ecorace are at the threshold of differentiating themselves. The high genetic variability present within wild Daba population of A. mylitta is of much importance for conservation as well as utilization in systematic breeding program.

Influence of thyroxine on different ion-dependent ATPase activities in fat body of tasar silkworm, Antheraea mylitta D.

The activities of Na(+)-K(+)-, Ca(2+)-, and Mg(2+)-ATPase of tasar silkworm, Antheraea mylitta D. fat body were investigated from fifth larval stage to adult emergence after injection of various doses (0.5, 1.0, 2.0, and 5.0 micrograms/g) of mammalian thyroxine (T4) to 1-hr-old fifth instar larvae. In normal silkworms, both sexes exhibited maximum enzyme activity before spinning (Day 12). Na(+)-K(+)-, Ca(2+)-, and Mg(2+)-ATPase activities in fat body of silkworms markedly declined after pupation and more so on the 1-day-old adults. All doses of thyroxine treated on fifth instar larvae significantly altered ATPase activity in the larval, pupal, and adult stages in both sexes. ATPase activity was not altered by lower dose of T4 (0.5 micrograms/g) in 2-day-old fifth stage larvae, while the higher dose (5.0 micrograms/g) surprisingly caused a reduction in ATPase activity during the different developmental stages. The fat body ATPases were influenced by thyroxine in all the stages of silkworm in a dose-dependent manner. Our results thus indicate that thyroxine has a controlling influence on the ATPase system in silkworm fat body.

L-thyroxine (T4) elevates the free amino acid pool of haemolymph plasma of tasar silkworm, Antheraea mylitta drury (Lepidoptera: Saturniidae).

Concentration of free amino acids (FAA) in the haemolymph plasma of male and female tasar silkworm, A. mylitta, during fifth larval instar were determined by amino acid analyzer after administration of vertebrate thyroxine (T4). Twenty FAA have been identified in haemolymph plasma in both sexes, out of which ten were found to be predominating. Treatment with anabolic doses of thyroxine (0.5 and 1.0 micrograms/g) were able to enhance the titre of all the individual FAA which has been also reflected in the total concentrations. A specific pattern of variations in plasma FAA recorded during 5th larval instar, was not altered by the administered thyroxine. Our results thus indicate that T4 has a controlling influence on the FAA profiles of silkworm as observed in other biochemical parameters of the insect.