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Abstract Background The choice of de novo assembler for high-throughput sequencing (HTS) data remains a pivotal factor in the HTS-based discovery of viral pathogens. This study assessed de novo assemblers, namely Trinity, SPAdes, and MEGAHIT for HTS datasets generated on the Illumina platform from 23 apple samples, representing 15 exotic and indigenous apple varieties and a rootstock. The assemblers were compared based on assembly quality metrics, including the largest contig, total assembly length, genome coverage, and N50. Results MEGAHIT was most efficient assembler according to the metrics evaluated in this study. By using multiple assemblers, near-complete genome sequences of citrus concave gum-associated virus (CCGaV), apple rubbery wood virus 1 (ARWV-1), ARWV-2, apple necrotic mosaic virus (ApNMV), apple mosaic virus, apple stem pitting virus, apple stem grooving virus, apple chlorotic leaf spot virus, apple hammerhead viroid and apple scar skin viroid were reconstructed. These viruses were further confirmed through Sanger sequencing in different apple cultivars. Among them, CCGaV, ARWV-1 and ARWV-2 were recorded from apples in India for the first time. The analysis of virus richness revealed that ApNMV was dominant, followed by ARWV-1 and CCGaV. Moreover, MEGAHIT identified novel single-nucleotide variants. Conclusions Our analyses highlight the crucial role of assembly methods in reconstructing near-complete apple virus genomes from the Illumina reads. This study emphasizes the significance of employing multiple assemblers for de novo virus genome assembly in vegetatively propagated perennial fruit crops.

Abstract Background To assess satisfaction level of patients undergoing septorhinoplasty pre- and post-operatively using Rhinoplasty Outcome Evaluation (ROE) questionnaire. Methods The present prospective study, after approval by Institutional Ethics Committee, was conducted in the Department of ENT, SMGS Hospital, from March 2021 to November 2022 on 83 patients who underwent septorhinoplasty. Demographic and clinical characteristics (age, gender, education, technique of surgery) of all participants were recorded. All patients were asked to answer the Rhinoplasty Outcome Evaluation (ROE) questionnaire 1 day prior to surgery. Postoperative ROE questionnaire was asked at the 6th month in ENT OPD by the same researcher. Results In our study, the mean preoperative total ROE score was 32.17 ± 5.38, and the mean postoperative total ROE score was 87.11 ± 5.39, the difference being statistically significant (p = 0.0039). Higher post-operative satisfaction score (89.7 ± 5.64) was seen in patients aged more than 35 years, as compared to younger patients (78.4 ± 5.8). In our study, upon gender-wise comparison, there was more statistically significant satisfaction in male patients than female patients postoperatively (p = 0.0022). Better postoperative ROE score was seen in lower literacy group (88.9 ± 6.73) than higher literacy group (83.3 ± 5.63). However, both open and closed techniques showed statistically insignificant difference in postoperative ROE score (p = 0.388). Conclusion Based on the Rhinoplasty Outcome Evaluation (ROE) questionnaire, our study showed that septorhinoplasty significantly improved quality of life in majority of our study subjects in terms of both aesthetic as well as functional parameters.

Abstract Background Where there is no identifiable cause of otologic symptoms like hearing loss and tinnitus, it is believed that etiology could possibly involve a vascular loop in AICA compressing the vestibulo-cochlear nerve within the internal auditory canal. In this study, we aim to evaluate whether there is any association of AICA vascular loops with unexplained tinnitus. Methods The present prospective study was conducted in the Department of ENT, SMGS Hospital, GMC Jammu, from October 2020 to March 2022 on 131 subjects with unexplained tinnitus. All subjects were subjected to contrast enhanced MRI brain (3D fast spin echo T2W1 with drive equilibrium pulse) for evaluation of AICA using Chavda classification—type 1 (lying in CP angle, but not entering IAC), type 2 (entering IAC but extending < 50% of length of IAC), and type 3 (extending > 50% of length of IAC). Results Out of 131 unexplained tinnitus cases, 76 patients (58%) had AICA loop on MRI. Out of 16 patients with right sided tinnitus, 11 patients had AICA on same side, while 5 patients had AICA on the opposite (left) side. Out of 26 patients with left-sided tinnitus, 12 patients had AICA on same side, while 14 patients had AICA on the opposite (right) side, these findings being statistically insignificant (p = 0.153). Conclusion From our study, we can conclude that the presence of AICA vascular loop either on CP angle or within IAC in tinnitus patients is an incidental finding and has no role in its etiology.

Abstract Pediatric liver transplantation rejection affects 20% of children. Currently, liver biopsy, expensive and invasive, is the best method of diagnosis. Discovery and validation of clinical biomarkers from blood or other biospecimens would improve clinical care. For this study, stored plasma samples were utilized from two cross-sectional cohorts of liver transplant patients at Children’s Healthcare of Atlanta. High resolution metabolic profiling was completed using established methods. Children with (n = 18) or without (n = 25) acute cellular rejection were included in the analysis (n = 43 total). The mean age of these racially diverse cohorts ranged from 12.6 years in the rejection group and 13.6 years in the no rejection group. Linear regression provided 510 significantly differentiating metabolites between groups, and OPLS-DA showed 145 metabolites with VIP > 2. A total of 95 overlapping significant metabolites between OPLS-DA and linear regression analyses were detected. Pathway analysis (p 500 significant metabolites. This result suggests that development of a non-invasive biomarker-based test is possible for rejection screening.

Geminiviruses are known to infect several fields and horticultural crops around the globe. Grapevine geminivirus A (GGVA) was reported in the United States in 2017, and since then, it has been reported in several countries. The complete genome recovered through high-throughput sequencing (HTS)-based virome analysis in Indian grapevine cultivars had all of the six open reading frames (ORFs) and a conserved nonanucleotide sequence 5′-TAATATTAC-3′ similar to all other geminiviruses. Recombinase polymerase amplification (RPA), an isothermal amplification technique, was developed for the detection of GGVA in grapevine samples employing crude sap lysed in 0.5 M NaOH solution and compared with purified DNA/cDNA as a template. One of the key advantages of this assay is that it does not require any purification or isolation of the viral DNA and can be performed in a wide range of temperatures (18°C–46°C) and periods (10–40 min), which makes it a rapid and cost-effective method for the detection of GGVA in grapevine. The developed assay has a sensitivity up to 0.1 fg μl-1 using crude plant sap as a template and detected GGVA in several grapevine cultivars of a major grapevine-growing area. Because of its simplicity and rapidity, it can be replicated for other DNA viruses infecting grapevine and will be a very useful technique for certification and surveillance in different grapevine-growing regions of the country.

Objectives: The bone quantity and quality determine the prosthetic success outcome. This research was performed to evaluate the bone density for insertion of pterygoid implants in edentulous and dentulous participants with cone-beam computed tomography (CBCT). Materials and Methods: CBCT evaluation was done for 66 dentate and edentulous patients for pterygoid implants at the pterygomaxillary region. The calculation of joint width, height, and volume of bone was done. Density of the bone was evaluated at the superior and inferior aspects of the pterygomaxillary column. Results: It was observed that average pterygomaxillary joint height for dentulous (dentate) was −12.7 ± 7.2 mm, edentulous −12.4 ± 7.1 mm, the average pterygomaxillary joint width for dentulous was 8.15 ± 7.3 mm, and 8.13 ± 6.2 mm for edentulous. The average pterygomaxillary joint volume in dentulous participants was 279.4 ± 189.2 mm3 and for edentulous was 254.5 ± 176.4 mm3. There was expressively greater density of the bone in dentulous participants over edentulous participants (P < 0.05). Conclusion: There was better bone density found in dentate participants in comparison to edentulous participants. CBCT is a recent investigative device which measures pterygoid area efficiently. Pterygoid implants may be deliberated as an alternative method for resorbed (atrophic) maxilla.

Objectives: Many authors' have advocated a minimum of two implants to support a mandibular overdenture, but financial restraints specifically among the geriatric population in developing countries made this treatment plan economically difficult. Hence, this study was planned to assess the symphyseal (midline) single implant-assisted complete overdenture for patient satisfaction and masticatory performance. Materials and Methods: In this clinical study, 12 edentulous first-time denture wearers underwent placement of a single implant in the mandibular symphyseal region. After 1 week, new complete dentures were fabricated and delivered to the patients. Post 3 months, the denture was fixed with a nylon cap-ball attachment to the anchor implant. Patients were questioned about comparison in the level of satisfaction and complaint before loading the implant (control group) and after 1 week, 1 month, and 3 months. The implant-assisted overdenture was fabricated with the help of a questionnaire. Masticatory performance was calculated with the help of a bite force measuring device at the same time intervals. SPSS 17.0 statistical software was used to analyze the data. Results: It was found that single implant anchorage of the mandibular complete denture resulted in a significant increase (P < 0.05) in patient's subjective satisfaction and a decrease (P < 0.05) in complaints at the end of 3 months. There was a significant (P < 0.01) increase in bite force in implant overdenture after 3 months (5.459 kgf) as compared to that of the complete denture (3.406 kgf). Conclusion: Single implant-assisted overdenture can be an appropriate treatment modality to treat edentulousness in the geriatric population. It insinuates the remarkable improvement of prosthesis function and oral comfort with minor surgical procedures.

Many youths participate in sports, and it is of interest to understand the impact of youth sports participation on later-life outcomes. However, prospective studies take a long time to complete and retrospective studies may be more practical and time-efficient to address some questions. We pilot a retrospective survey of youth sports participation and examine agreement between respondent's self-reported participation with high school records in a sample of 84 adults who graduated from high school between 1948 and 2018. The percent agreement between our survey and the school resources for individual sports ranged between 91.5% and 100%. These findings provide preliminary evidence for the reliability of retrospective self-report of youth sports participation. This survey may serve as an efficient approach for evaluating relationships between involvement in youth sports and health outcomes later in adulthood.

The pathophysiology of periodontal disease involves a perturbed immune system to a dysbiotic microflora leading to unrestrained inflammation, collateral tissue damage, and various systemic complications. Gingival epithelial cells function as an important part of immunity to restrict microbial invasion and orchestrate the subsequent innate responses. A20 (TNFAIP3), an ubiquitin-editing enzyme, is one of the key regulators of inflammation and cell death in numerous tissues including gastrointestinal tract, skin, and lungs. Emerging evidence indicates A20 as an essential molecule in the oral mucosa as well. In this study, we characterized the role of A20 in human telomerase immortalized gingival keratinocytes (TIGKs) through loss and gain of function assays in preclinical models of periodontitis. Depletion of A20 through gene editing in TIGKs significantly increased IL-6 and IL-8 secretion in response to Porphyromonas gingivalis infection while A20 over-expression dampened the cytokine production compared to A20 competent cells through modulating NF-κB signaling pathway. In the subsequent experiments which assessed apoptosis, A20 depleted TIGKs displayed increased levels of cleaved caspase 3 and DNA fragmentation following P. gingivalis infection and TNF/CHX challenge compared to A20 competent cells. Consistently, there was reduced apoptosis in the cells overexpressing A20 compared to the control cells expressing GFP further substantiating the role of A20 in regulating gingival epithelial cell fate in response to exogenous insult. Collectively, our findings reveal first systematic evidence and demonstrate that A20 acts as a regulator of inflammatory response in gingival keratinocytes through its effect on NF-κB signaling and desensitizes cells to bacteria and cytokine induced apoptosis in the oral mucosa. As altered A20 levels can have profound effect on different cellular responses, future studies will determine whether A20-targeted therapies can be exploited to restrain periodontal inflammation and maintain oral mucosa tissue homeostasis.

Adverse clinical events in primary sclerosing cholangitis (PSC) happen too slowly to capture during clinical trials. Surrogate endpoints are needed, but no such validated endpoints exist for children with PSC. We evaluated the association between gamma glutamyltransferase (GGT) reduction and long‐term outcomes in pediatric PSC patients. We evaluated GGT normalization (< 50 IU/L) at 1 year among a multicenter cohort of children with PSC who did or did not receive treatment with ursodeoxycholic acid (UDCA). We compared rates of event‐free survival (no portal hypertensive or biliary complications, cholangiocarcinoma, liver transplantation, or liver‐related death) at 5 years. Of the 287 children, mean age of 11.4 years old, UDCA was used in 81% at a mean dose of 17 mg/kg/day. Treated and untreated groups had similar GGT at diagnosis (314 versus 300, P= not significant [NS]). The mean GGT was reduced at 1 year in both groups, with lower values seen in treated (versus untreated) patients (99 versus 175, P= 0.002), but 5‐year event‐free survival was similar (74% versus 77%, P= NS). In patients with GGT normalization (versus no normalization) by 1 year, regardless of UDCA treatment status, 5‐year event‐free survival was better (91% versus 67%, P< 0.001). Similarly, larger reduction in GGT over 1 year (> 75% versus < 25% reduction) was also associated with improved outcome (5‐year event‐free survival 88% versus 61%, P= 0.005). Conclusion:A GGT < 50 and/or GGT reduction of > 75% by 1 year after PSC diagnosis predicts favorable 5‐year outcomes in children. GGT has promise as a potential surrogate endpoint in future clinical trials for pediatric PSC.

Background and Aims: Oncosurgeries may incur massive blood loss demanding frequent blood sampling to assess blood loss and the need for intraoperative blood transfusions. Accuracy of non-invasive spectrophotometric haemoglobin (hereafter to be referred as SpHb) monitoring has been studied in various perioperative settings. The intraoperative use of Radical-7®, Masimo Corp., (Radical-7®) for SpHb monitoring may be useful during cancer surgery. The aim of this study is to evaluate the intraoperative utility of SpHb monitoring by the Radical-7® to guide intraoperative transfusion in oncosurgeries. Methods: Fifty adult patients, undergoing oncosurgery with anticipated blood loss of more than 20% of blood volume, were selected. Continuous SpHb monitoring was performed intraoperatively and blood transfusion was based on SpHb values. Simultaneous laboratory haemoglobin (LabHb) samples were taken for validation. The accuracy of intraoperative blood transfusions based on SpHb was analysed using Error Grid Analysis. Paired measurements of SpHb and LabHb were compared using Bland–Altman plot analysis. Results: There were 66 paired data points for blood transfusion from fifty patients with a correlation of 73% (P < 0.001) between SpHb and LabHb. In the Bland–Altman analysis, the bias was − 0.313 g/dl with ~ 95% of values within the limits of agreement of 1.81 g/dl to −2.44 g/dl. In the Error Grid Analysis, most data points were in the least error zone (Zone A). Conclusion: The Radical-7® has the advantage of providing SpHb value continuously to take prompt decision regarding blood transfusion intraoperatively.

Objective: This study aims to study the difference in etiology and outcome in terms of implantation rate and abortion rate in fresh (self-stimulated) versus frozen (oocyte donation cycle) in vitro fertilization (IVF) and in transient versus persistent fluid. Material and Methods: This retrospective study was conducted in the Department of Reproductive Medicine of tertiary care center from January 2012 to November 2015. Data were collected retrospectively from the departmental files. Twenty-four patients from fresh IVF-stimulated cycles and 24 from frozen oocyte donation cycle with their endometrium prepared by hormone replacement treatment were included in the study. All patients selected in the study had grade-A embryo transfer of day 3–4 with maximum three embryo transferred. Pregnancy was defined by rising serum beta-human chorionic gonadotrophin levels performed after 14 days of embryo transfer and further confirmed by ultrasonographic visualization of gestational sac at 6 weeks. All biochemical pregnancies were included in implantation failure. All pregnant patients were followed till the termination of pregnancy and further noted as live birth or abortion. Results: Clinical pregnancy rate was seen more in self-stimulated cycle (62.5%) with live birth rate of 50% than hormone replacement treatment cycle, in which clinical pregnancy rate was 45.83% with live birth rate of 33.33%. Clinical pregnancy rate was highest in group with very less fluid in cavity (1–2 mm) 63% and with live birth of 52.63%. Clinical pregnancy was seen only in two patients of group B with anterior and posterior (AP) diameter of fluid in cavity of 2–3 mm with live birth of only one, whereas in group C, with AP diameter of 3–5 mm, none of the patient conceived. This difference was statistically significant. Clinical pregnancy rate was 65.62% in transient fluid accumulation with live birth rate of 53.25%, which was significantly higher than persistent fluid accumulation (P value − 0.0337 for pregnancy rate and 0.0312 for live birth rate). Conclusion: Fluid accumulation seen in fresh cycles are generally associated with better outcome because it may be associated with good prognostic factors – small AP diameter of fluid, with transient fluid accumulation and more with poly cystic ovarian syndrome as an etiological factor; however, in frozen cycle, it can be associated with poor outcome.

The use of implants in the oral and maxillofacial skeleton continues to expand and since then implant surfaces have been modified in various ways to improve biocompatibility and accelerate osseointegration, which results in a shorter edentulous period for a patient. This article reviewed several methods that are widely used to modify the topography or chemistry of titanium surface, including blasting, acid etching, anodic oxidation, fluoride treatment, and calcium phosphate coating. Such modified surfaces demonstrate faster and stronger osseointegration than the turned commercially pure titanium surface.

CONTEXT: Fibroids are the most common tumors of the uterine cavity. Most of them are diagnosed during the reproductive age when the fertility is an important concern for the female. However, complications can occur after removal of fibroid (myomectomy) too for future pregnancy. Though myomectomy has been sighted as a cause of intrauterine adhesions data regarding the effect of myomectomy on endometrial cavity is lacking. AIMS: Evaluate the incidence of intrauterine adhesion formation after myomectomy and to identify the associated factors. MATERIALS AND METHODS: In this prospective observational study, hysteroscopy was done in 51 infertile patients who had undergone myomectomy 3 months before in a tertiary care center from 2012 to 2015. The presence of intrauterine adhesions noted on hysteroscopy was investigated on the basis of size, number, location and type of fibroid removed, along with intraoperative breach of the uterine cavity. STATISTICAL ANALYSIS: Chi-square test was used for the calculating significant difference in frequency of discrete variables in two groups. P < 0.05 was considered significant. RESULTS: Intrauterine adhesions were seen in 11 out of 51 (21.57%) cases. No significant relationship between intrauterine adhesions and type, size or number of fibroid was observed. No statistical difference in the rate of adhesion formation was seen irrespective of breach of the uterine cavity during myomectomy. CONCLUSION: Intrauterine adhesion formation after myomectomy is not related to the type of surgery or the nature of fibroid. However, in all cases desiring fertility postoperative hysteroscopy is highly recommended to diagnose and treat these adhesions early.

INTRODUCTION: This study was undertaken to compare between clomiphene citrate (CC) and gonadotropin-releasing hormone antagonist-based protocols in donor-recipient cycles in terms of parameters of ovarian stimulation and obstetric outcome. MATERIALS AND METHODS: Two hundred and three fertile oocyte donors were stimulated using two different protocols: Clomiphene based (n = 103) and antagonist based (n = 100). Donors in the one group were stimulated from day 1 or 2 of spontaneous or withdrawal bleeds with CC (50 mg/day) and recombinant follicle-stimulating hormone (FSH) till the day of trigger while donors in the other group were stimulated using recombinant FSH from day 1 or 2, and the antagonist was added as per flexible antagonist protocol. When >3 follicles were >17 mm in diameter, trigger was given with 2 mg leuprolide intramuscular. Transvaginal oocyte retrieval was done after 34 h of trigger. RESULTS: There was no significant difference in between the two groups in terms of age, antral follicle count, starting dose of gonadotropins, total dose required, duration of stimulation, number of follicles retrieved, mature follicles, and fertilization rate. The serum estradiol levels were significantly raised in the clomiphene group (P < 0.001). Pregnancy rate was similar in both the groups. The clinical pregnancy rate was 65.94% in the clomiphene group and 57.46% in the antagonist group. The live birth rate per cycle started was 47.8% in the clomiphene group and 39.55% in the antagonist group. There was one case of ectopic pregnancy in the antagonist group. CONCLUSION: Controlled ovarian stimulation using clomiphene and gonadotropin is a viable option for donor oocyte cycles. The cost and number of injections used per cycle can be reduced by using the clomiphene-based protocols.

INTRODUCTION: With the advent of assisted reproductive treatment options, the incidence of multiple pregnancies has increased. Although the need for elective single embryo transfer is emphasized time and again, its uniform applicability in practice is yet a distant goal. In view of the fact that triplet and higher order pregnancies are associated with significant fetomaternal complications, the fetal reduction is a commonly used option in such cases. This retrospective study aims to compare the perinatal outcome in patients with triplet gestation who have undergone spontaneous fetal reduction (SFR) as against those in whom multifetal pregnancy reduction (MFPR) was done. MATERIALS AND METHODS: In the present study, eighty patients with triplet gestation at 6 weeks were considered. The patients underwent SFR or MFPR at or before 12-13 weeks and were divided into two groups (34 and 46), respectively. RESULTS: Our study found no statistical difference in perinatal outcome between the SFR and MFPR groups in terms of average gestational age at delivery, abortion rate, preterm delivery rate, and birth weight. The study shows that the risk of aborting all fetuses after SFR is three times (odds ratio [OR] = 3.600, 95% confidence interval [CI] = 0.2794-46.388) that of MFPR in subsequent 2 weeks. There were more chances of loss of extra fetus in SFR (23.5%) group than MFPR group (8.7%) (OR = 3.889, 95% CI = 1.030-14.680). As neither group offers any significant benefit from preterm delivery, multiple pregnancies continue to be responsible for preterm delivery despite fetal reduction. CONCLUSION: There appears to be some advantages of MFPR in perinatal outcome when compared to SFR, especially if the latter happens at advanced gestation. Therefore, although it is advisable to wait for SFR to occur, in patients with triplet gestation at 11-12 weeks, MFPR is a viable option to be considered.

Introduction. Aim of the study was to find the effect of various prognostic factors in cases of unexplained infertility undergoing controlled ovarian stimulation (COS) with intrauterine insemination (IUI). Methods. 146 cases of unexplained infertility were included. A maximum of 3 cycles of IUI were done with clomiphene citrate/HMG. Ovulation trigger was given when the largest follicle diameter was >18 mm, and IUI was planned 36 hours later. Luteal phase support was given for 15 days, urine pregnancy test was done on day 15, ultrasonography was done at 7 weeks, and pregnancy was followed up till delivery. Results. A total of 146 couples have undergone 239 cycles of IUI out of which 27 had UPT positive after 15 days. 14.8% had 1st-trimester abortion while 3.7% were ectopic. 86.3% were singleton pregnancies and 13.6% were twins. CPR was 11.29% per cycle and 18.4% per couple; LBR was 9.2% per cycle. Apart from duration of stimulation (p=0.037) and number of treatment cycles (p=0.045), no other factors had significant prognostic value. Conclusion. For unexplained infertility, IUI can be done to provide patients with the time that they need before moving on to IVF while providing a respectable chance of pregnancy.

Objectives. This study aims to evaluate perinatal outcomes such as gestational age at delivery and live birth rate in singleton and twin gestation with or without fetal reduction. Method. A retrospective analysis was done on patients which were divided into reduced and unreduced groups on the basis of order of reduction of one or more fetuses between 6 and 13 weeks of gestation. Patients records were studied to note gestational age at delivery/abortion, birth weight, and neonatal outcome. Result. The cohort included a total of 292 patients: 102 singletons and 190 twins. 52 pregnancies were reduced in singleton cohort and 68 were reduced in twin cohort. No statistical difference was observed in live birth rate, gestational age at delivery, and birth weight and significant higher incidence of IUGR was observed in reduced and unreduced twin gestation. In singleton pregnancies however preterm delivery rate increased with fetal reduction. Conclusion. Although reduction does not reduce the live birth rate, it does reduce gestation age of delivery and birth weight of newborn. This effect is more apparent when multiple gestation is reduced to singleton.