A Study on the Outcome of Myringoplasty in Small to Medium Sized Perforations Using Collagen Sheet as Graft Material.

Chronic Otitis Media is a global health problem causing significant morbidity of life. Various medical and surgical treatment modalities are being used for the treatment of Chronic Otitis Media. Surgical treatment is the definitive option which includes different surgical procedures, all of which have been designed to attain the same goals of eradicating disease and achieve a safe and dry ear with closure of tympanic membrane perforations. Various graft materials are being used for the closure of tympanic membrane perforations like autografts, isografts, allografts and xenografts, each having its own advantages and limitations. The trend towards increasingly less invasive medical procedures, with shorter hospitalisation stay has led to demand for materials to replace autologous grafts, one such material being the collagen. The purpose of this study is to evaluate the outcome of myringoplasty in small to medium sized perforations using collagen sheet as the graft material. In the period from November 2018 to May 2020, 50 patients after fulfilling the inclusion and exclusion criteria were taken up for myringoplasty. They underwent myringoplasty using transcanal approach and collagen sheet was used to close the tympanic membrane perforation. Post-operatively, patients were treated with antibiotics and regularly followed up. Otoendoscopy was done at the end of 1st month and 3rd month. At the end of 3 months, patients were evaluated for graft uptake and pure tone audiometry repeated to evaluate the post-operative hearing gain. In this study, we examined a total of 50 patients with 25 males and 25 females (50%). The mean age of our patients was 29.42 years, ranging from 18 to 60 years; with almost 66% between 21 and 40 years. Treatment was considered successful if there was complete closure of tympanic membrane perforations. Of the 50 patients that were included; graft uptake was noted in 46 patients i.e. in 92%, closure was achieved at the end of 3 months. Four failures in our study were due to infections. In this study, the graft uptake following myringoplasty was analyzed with statistically significant objective parameters like age, gender, size of perforation, laterality of disease, etiology of CSOM that affect the success of tympanoplasty. It was observed that small sized perforations had more chances of graft uptake. Post-operative hearing outcomes were also evaluated and was found to be statistically significant. Collagen sheet provided promising results as a graft material for myringoplasty and the graft uptake rates were comparable to the success rates of other conventional autologous grafts. Post operatively, significant improvement in hearing was also noted.

Transcriptomic Variation of Amphiprion Percula (Lacepède, 1802) in Response to Infection with Cryptocaryon Irritans Brown, 1951.

Cryptocaryon irritans (Brown 1951) frequently infect the Pomacentridae fishes causing severe economic losses. However, the anti-C. irritans' molecular mechanism in these fishes remains largely unknown. To address this issue, we conducted RNA-Seq for C. irrtians-infected gills of the clownfish Amphiprion percula (Lacepède 1802) at the early (day 1) and late (day 3) stages of infection. A total of 1655 differentially expressed genes (DEGs) were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of DEGs showed a vast genetic variation related to the following aspects: ECM-receptor interaction, P13K-Akt signalling, cytokine-cytokine receptor interaction, and endocytosis. During the early phase of infection, key genes involved in ATP production, energy homeostasis, and stress control were abruptly increased. In the late phase, however, acute response molecules of the peripheral nervous system (synaptic transmission and local immunity), metabolic system triggering glycogen synthesis, energy maintenance, and osmoregulation were found to be critical. The highest number of upregulated genes (URGs) recovered during the early phase was included under the 'biological process' category, which primarily functions as response to stimuli, signalling, and biological regulation. In the late phase, most of the URGs were related to gene regulation and immune system processes under 'molecular function' category. The immune-related URGs of early infection include major histocompatibility complex (MHC) class-II molecules apparently triggering CD4+ T-cell-activated Th responses, and that of late infection include MHC class-1 molecules for the possible culmination of CD8+ T-cell triggered cytotoxicity. The high level of genic single nucleotide polymorphisms (SNPs) identified during the late phase of infection is likely to influence their susceptibility to secondary infection. In summary, the identified DEGs and their related metabolic and immune-related pathways and the SNPs may provide new insights into coordinating the immunological events and improving resistance in Pomacentridae fishes against C. irritans.

Benign acute childhood myositis following human Influenza B virus infection - A case series.

Influenza virus primarily affects ciliated cells of respiratory epithelium. Humans do not have innate immunity for these viruses and are vulnerable to get attacked. Benign acute childhood myositis usually occurs at the early convalescent phase of a influenza viral illness when fever, cough, myalgia, nasal discharge are the initial presentation.

The effect of chlorhexidine gluconate chips on clinical status of peri-implant mucosa and plaque formation on healing abutments following single-stage implant surgeries.

Background: An experimental study was designed to evaluate the effect of chlorhexidine gluconate chips on clinical status of peri-implant mucosa and plaque formation on healing abutments following single-stage implant surgeries. Materials and Methods: Twenty-eight single-stage implant sites were grouped into 14 test and control sites. The study commences from the time of suture removal following surgery, designated as day 0. Chlorhexidine chip insertion into peri-implant sulcus in test sites was done on day 0, 10, and 20. Peri-implant crevicular fluid was collected on day 0, 10, 20, and 30 for biochemical estimation of aspartate aminotransferase (AST). The modified sulcus bleeding index (mBI) score was obtained in both test and control sites on day 10, 20, and 30. On day 30, all the healing abutments were unscrewed and sent for stereomicroscopic analysis to assess the plaque formation on its surface. Results: Statistically significant difference was not observed in AST levels and mBI in both test and control groups in various time intervals. In the stereomicroscopic assessment of healing abutment, mild grade of plaque accumulation was seen in three samples in test group, one sample in control group, and severe grade was seen in six samples in test group and nine samples in the control group. Conclusion: Inflammatory condition of peri-implant mucosa and plaque retentive properties on healing abutment surface were found to have reduced with the usage of chlorhexidine gluconate chips. However, the study failed to establish a statistically significant correlation of these observations.

Dengue and scrub typhus co-infection in children: Experience of a teaching hospital in an endemic area.

Background: Dengue fever and scrub typhus are considered an endemic disease in the Indian subcontinent. The epidemiology and clinical presentations are complex and vary each year. Objective: The objective of this study was to estimate the prevalence of coinfection with scrub typhus in children diagnosed with dengue fever. Methods: A retrospective hospital-based, cross-sectional study was done in the Department of Pediatrics of a teaching hospital in Puducherry. All children (0-14 years) who had enzyme-linked immunosorbent assay (ELISA) reported scrub typhus among those diagnosed with dengue fever (NS1Ag or immunoglobulin M ELISA positivity) during 2012-2016. Medical records with incomplete data were excluded from the study. Odds ratio was calculated to find out the association of coinfections. An independent t-test was used to find out the statistical significance. P < 0.05 was considered statistically significant. Results: Atypical features of dengue were present in 250/318 (78.6%) children. Coinfections were seen in 62/318 (19.4%) children. Scrub typhus was the most common (n = 51/62, 82.2%). The chance of scrub typhus in a dengue serology-positive child is significant when the symptoms are atypical or protracted (OR- 2.6, P = 0.033). Conclusion: High index of suspicion should be present in endemic dengue and scrub typhus coinfection.

Modern biotechnological strategies for vaccine development in aquaculture - Prospects and challenges.

Advances in genomics and the gradual reduction of cost for technologies like whole-genome sequencing have provided exciting opportunities for developing modern biotechnological-based vaccines in aquaculture. This systemic review describes the prospects and challenges of implementing these high-tech vaccines in fish species. The majority of the commercial vaccines in aquaculture utilize conventional procedures for which cost of administration, protective immunity and safety issues are the major challenges. In recent years, more efficient vaccines are being developed by adopting the advances in vaccine technology. Vaccines based on surface antigens, protein/peptide/polysaccharide subunits, recombinant DNA/mRNA/plasmids, novel antigen expression and delivery systems (bacteriophage particles, virus like particles/VLPs, recombinant yeast, mucosal vaccines), novel molecular adjuvants (IL-8, IL-12, HSPs), and encapsulation polymers and polysaccharides like chitosan nanoparticles and PLGA microcapsule were successfully developed. These biotechnology-based vaccines have proved to be very efficient in field trials, but are always in the research pipeline or as patents. Only very few of them are licensed for use, that too, in high-valued fishes like salmonids. Currently, commercial aquaculture vaccines are available for Aeromonas salmonicida, Vibrio salmonicida, Yersinia ruckeri, Vibrio anguillarum, Edwardsiella ictalurid, and for certain Betanodaviruses. Nevertheless, no registered vaccines are available for other major infectious diseases/pathogens such as viral hemorrhagic septicemia virus (VHSV), viral nervous necrosis virus (VNN) and certain other betanodaviruses, channel catfish virus (CCV), gill disease bacteria, mycobacteria, flavobacterium, Edwardsiella tarda, and certain streptococci. Despite the important economic losses that the pathogens cause to aquaculture worldwide, the commercialization of vaccines remains limited due to immunological pitfalls in aquatic species, large-scale vaccination issues, unregulated use of antibiotics and chemicals, gene-based vaccine regulations and commercial viability. If attempts are to be made to develop novel delivery methods, cost-effective procedures, and relaxations in DNA vaccine regulations, biotechnology-based vaccination could circumvent the emerging disease challenges in aquaculture.

Immune responses of Cyprinus carpio induced by protein extracts of Lernaea cyprinacea Linnaeus, 1758.

Lernaea cyprinacea Linnaeus, 1758 is an ectoparasite showing widespread infections in tropical aquaculture, and the present study aimed to determine the specific immune responses against this parasite. For the experiment, whole parasite extracts were injected intraperitoneally into Cyprinus carpio Linnaeus, 1758, and samples of epidermal mucus and blood were drawn at 0, 1, 7 and 14-days post-injection (DPI). The results revealed high levels of protein, protease and lysozyme activities in the experimental fish which were injected with L. cyprinacea protein extract. In the epidermal mucus, the total protein concentration of the control fish was 460 µg/mL, and the level raised significantly to 800 µg/mL in the experimental fish. The lysozyme activity increased from 741.5 u/mL to a peak level of 1448.5 u/mL at 7DPI. The protease activity was also found elated gradually from 2.91 u/µL to 4.49 u/µL at 1 to 14 DPI. In the serum samples, the protein concentration remained steady throughout the experiment period. However, all the experimental fish displayed statistically high levels of lysozyme and protease activity, from 890 u/mL to 1220 u/mL, and 6.10 u/µL to 11.88 u/µL, respectively. In the whole blood samples, the haemoglobin content and the red blood cells (RBC) count did not show any significant change in any of the experimental groups. But, the percentage of lymphocytes showed a marginal increase from 0.47 to 0.6 in the experimental groups. Overall, the immune responses induced by L. cyprinacea protein extracts depicts a pattern of specific responses, in which the local humoral responses dominate the systemic humoral/cellular response. The results further revealed the possibility of futuristic approaches to control freshwater ectoparasites.

Comparative Evaluation of Different Soft Tissue Coverage Techniques at Immediate Implant Sites: A Cohort Study.

AIM AND OBJECTIVE: To compare the mucosal thickness, width of attached gingiva, and extent of coverage achieved with the usage of connective tissue graft, platelet-rich fibrin (PRF) membrane and buccally advanced flap along with bone grafts in the closure of immediate implant site. MATERIALS AND METHODS: Twenty-one sites requiring immediate implants were randomly divided into three groups of seven samples each. The techniques comprised bone grafting in jumping space along with either buccally advanced flap (group A), PRF membrane (group B), or connective tissue graft (group C) to cover the socket. In each group, the extent of socket coverage, mucosal phenotype, and width of attached mucosa achieved were assessed after 3 months and intercompared with initial measurements to identify the best technique in achieving primary tissue closure of immediate implant sites. RESULTS: Comparative assessment of gain in attached mucosal width had a remarkable difference in all the groups but exhibited no statistical significance among the groups compared. Also, comparative assessment of mucosal thickness and the extent of socket coverage equally indicated a clinical significance among all groups, but failed to achieve any statistical significance. CONCLUSION: All the three techniques were found to be equally effective in achieving additional width of attached mucosa around implants, coverage of mucosa at osteotomy sites, and a thicker mucosal phenotype at implant sites. In comparison with one another, no single technique was found to be advantageous over the other. CLINICAL SIGNIFICANCE: The commonly used three techniques for socket coverage following the immediate implant placement has been proven to be equally effective. Thus, the selection of the technique to meet the objective of complete closure of an extraction socket along with implant platform is left open to operator skill and operation site, which should consider least traumatic and most feasible technique.

Comparative evaluation of the effect of Ozone therapy and Photodynamic therapy in non-surgical management of Chronic periodontitis: A split mouth longitudinal study.

BACKGROUND: A split-mouth longitudinal study was conducted to compare and evaluate the effect of ozonated water and photodynamic therapy (PDT) in nonsurgical management of chronic periodontitis, along with mechanical debridement procedure. MATERIALS AND METHODS: Twenty-two patients diagnosed with chronic generalized periodontitis were subjected to the study. Following the assessment of gingival index, periodontal pocket depth, and clinical attachment loss, all patients underwent full-mouth scaling and root planing. Upper right and left quadrants of each patient were considered as sample sites in the study. Among these split-mouth sites, upper right quadrant of each patient was subjected to ozonated water irrigation with a 22-gauge needle and left upper quadrant was treated with PDT, which involved sulcus irrigation with indocyanine green dye (0.05 mg/ml) followed by low-level diode laser light application at 0.5 W and 810 nm (AMD Picasso) through a fiber-optic tip of 10 mm length, default angle of 60°, and fiber core diameter of 400 µm in noncontact continuous wave mode. Patients were recalled at the 2nd and 4th months regularly, and the therapy was repeated at the same sites in the same manner. Clinical parameters recorded before the study were assessed again at the end of the 2nd- and 6th-month period. RESULTS: A statistically significant reduction (P < 0.05) was observed in gingival index scores within both the study groups at all intervals of the study. In Ozone therapy (OT) group, a statistically significant difference was noted for total periodontal pocket depth values between baseline and 2nd month (P = 0.000), baseline and 6th month (P = 0.000), and between 2nd month and 6th month (P = 0.029). In the PDT group, on contrary, a statistically significant difference was noticed in total periodontal pocket probing depth values between baseline and 2nd month (P = 0.000) and baseline to 6th month (P = 0.000), but a similar significant difference was not noticed between 2nd-month and 6th-month periods (P = 0.269). In group OT, a statistically significant difference was noted for total clinical attachment loss between baseline and 2nd month (P = 0.000), baseline and 6 months (P = 0.000), and 2nd month and 6th month (P = 0.019). In group PDT, a statistically significant difference in terms of its improvement was noted at intervals between baseline and 2 months (P = 0.000) and from baseline to 6 months (P = 0.000) but not between 2nd month and 6th month (P = 0.129). CONCLUSION: Results of the study showed that sub-gingival OT and PDT equally improved the clinical outcomes of treatment drastically following mechanical debridement at the end of first 2 months. Thereafter, it was shown to improve steadily throughout the study period, with slightly better results with OT compared with PDT.

Assessment of the effect of ozonated water irrigation on gingival inflammation in patients undergoing fixed orthodontic treatment.

BACKGROUND: A split-mouth longitudinal study was designed here to evaluate the effect of ozonated water delivered subgingivally through a jet device (Kent Ozone dental TY 820 jet) in controlling gingival inflammation of patients undergoing fixed orthodontic therapy. MATERIALS AND METHODS: Twenty-eight patients undergoing fixed orthodontic treatment following thorough oral prophylaxis were recalled after 2 weeks, and baseline gingival inflammation was estimated. This study was conducted as split-mouth technique with the upper left quadrant (Control side) was irrigated by jetting saline into sulcus by air-water syringe with gauge 22 needle tip and upper right quadrant (Test side) irrigated with ozonated water by a jet device using similar delivery tip. The patients were recalled at the end of 1st, 2nd, 3rd, and 4th month and the irrigations as well as gingival inflammation estimation were repeated. A biochemical assessment of gingival inflammation (estimation of aspartate aminotransferase [AST] in gingival crevicular fluid [GCF]) and clinical assessment (gingival index scoring) was done at each visit. RESULTS: At the test side, the AST level in the GCF was found to be maintained almost the same value as baseline till the end of 4th month, indicating not much of change in inflammatory status. Statistically significant difference in GCF-AST and gingival index score between test and control sites were found at the end of 3rd and 4th month with a significance of P < 0.05. CONCLUSION: The patient undergoing fixed orthodontic treatment showed a consistent improvement in gingival inflammation on frequent irrigation with ozone jet irrigation as compared with saline irrigation delivered with same jetting force.

Transcriptomic analysis of the housefly (Musca domestica) larva using massively parallel pyrosequencing.

To explore the transcriptome of Musca domestica larvae and to identify unique sequences, we used massively parallel pyrosequencing on the Roche 454-FLX platform to generate a substantial EST dataset of this fly. As a result, we obtained a total of 249,555 ESTs with an average read length of 373 bp. These reads were assembled into 13,206 contigs and 20,556 singletons. Using BlastX searches of the Swissprot and Nr databases, we were able to identify 4,814 contigs and 8,166 singletons as unique sequences. Subsequently, the annotated sequences were subjected to GO analysis and the search results showed a majority of the query sequences were assignable to certain gene ontology terms. In addition, functional classification and pathway assignment were performed by KEGG and 2,164 unique sequences were mapped into 184 KEGG pathways in total. As the first attempt on large-scale RNA sequencing of M. domestica, this general picture of the transcriptome can establish a fundamental resource for further research on functional genomics.

Codon changed immobilization antigen (iAg), a potent DNA vaccine in fish against Cryptocaryon irritans infection.

The immobilization antigen (iAg) DNA sequence from Chiayi isolate of Cryptocaryon irritans was computationally reviewed to replace the stop codons with suitable amino acids and its GC content was intensified. The plasmid construct comprising the codon changed iAg (optiAg/optimized iAg) was successfully expressed in the bacterial strain BL21 and also in grouper fin cells (GF-1). Results of immobilization assay, ELISA and western blot of C. irritans theront and recombinant iAg by grouper antiserum against optiAg DNA indicated that the codon changed iAg retains the native conformation. The DNA vaccine construct pcDNA3.1-optiAg was encapsulated in water-oil-water triple layer emulsions measuring 19 µm diameters and was used for the immunization experiment. In trial I experiment, grouper fish were immunized twice via intramuscular injection with the pcDNA3.1-optiAg and were challenged with C. irritans at 8-day post immunization (dpi), which resulted in 46% relative percent survival (RPS). In trial II, single immunization with pcDNA3.1-optiAg boosted with recombinant iAg protein, resulted in 40% RPS. The data from this study reveal that codon change in iAg not only accomplished the expression of iAg protein in both prokaryotic and eukaryotic cell systems, but also optiAg was proved as immunogenic due to the protection it confers to the immunized fish against C. irritans infection. Hence, it is concluded that iAg can be a potent DNA vaccine in fish against infection of the ciliated protozoan, C. irritans.

Laparoscopic diagnosis of incarcerated "spigelian hernia": report of a case and review of the literature.

Spigelian hernias are rare hernias, representing only about 1-2% of all abdominal hernias. An accurate preoperative diagnosis of this condition is often difficult because the physical presence of these hernias is often not demonstrable owing to its peculiar anatomic location. Many patients with Spigelian hernias, therefore, may have no obvious findings on clinical examination. The diagnosis is even trickier in obese patients wherein the hernia sac may lie in an intraparietal plane, masked by the abdominal subcutaneous fat. Here we describe a case of incarcerated Spigelian hernia where clinical and radiological findings were inconsistent and the accurate diagnosis was made by laparoscopy and was followed by a laparoscopic-assisted repair using an open anterior approach with an onlay mesh. We therefore feel that laparoscopy can be a useful tool for confirming the diagnosis and also for performing a definitive repair in doubtful cases of Spigelian hernias.

A Toll receptor from Chinese shrimp Fenneropenaeus chinensis is responsive to Vibrio anguillarum infection.

Toll-like receptors (TLRs) are an evolutionarily ancient family of pattern recognition receptors (PRRs), playing a crucial role in innate immune responses. Here we present a Toll homolog from Chinese shrimp Fenneropenaeus chinensis, designated FcToll. The full-length cDNA of FcToll is 4115 bp including a poly A-tail of 16 bp, encoding a putative protein of 931 amino acids. The predicted protein consists of an extracellular domain with a potential signal peptide, 16 leucine-rich repeats (LRR), two LRR-C-terminal (LRR-CT) motifs, and two LRR-N-terminal (LRR-NT) motifs, followed by a transmembrane segment of 23 amino acids, and a cytoplasmic Toll/Interleukin-1R (TIR) domain of 139 residues. Genomic structure of FcToll gene contains five exons and four introns. Phylogenetic analysis revealed that it belongs to insect-type invertebrate Toll family. Transcripts of FcToll gene were constitutively expressed in various tissues, with predominant level in lymphoid organ. Real-time PCR assays demonstrated that expression patterns of FcToll were distinctly modulated after bacterial or viral stimulation, with significant enhancement after 5h post-Vibrio anguillarum challenge but markedly reduced levels immediately after white spot syndrome virus (WSSV) exposure. These results suggest that FcToll might be involved in innate host defense, especially against the pathogen V. anguillarum.