Transcriptome mining and expression analysis of ABC transporter genes in a monophagous herbivore, Leucinodes orbonalis (Crambidae: Lepidoptera).

Insecticide resistance is a global concern and requires immediate attention to manage dreadful insect pests. One of the resistance mechanisms adopted by insects is through ATP-binding cassette (ABC) transporter proteins. These proteins rapidly transport and eliminate the insecticidal molecules across the lipid membranes (Phase III detoxification mechanism). In the present study, we investigated the potential role of ABC transporter genes in imparting insecticide resistance in field-collected insecticide resistant larvae of eggplant shoot and fruit borer (Leucinodes orbonalis; Crambidae: Lepidoptera). Dose-mortality bioassays against five insecticidal molecules revealed moderate to high levels of insecticide resistance (32.2. to 134.1-fold). Thirty-one genes encoding ABC transporter proteins were mined from the transcriptome resources of L. orbonalis. They were classified under eight sub-families (ABCA to ABCH). Phylogenetic analysis indicated ABCG is the most divergent, composed of nine genes as compared to many other insects. Transcriptome analysis of the insecticide resistant and susceptible strains of L. orbonalis revealed differential expression of 13 ABC transporter genes. The altered expression of these genes was further validated using qRT-PCR. The knockdown studies indicated the involvement of ABCD1 and ABCG2 genes in chlorantraniliprole resistance in the insecticide-resistant strain of L. orbonalis. This study unveils the additional mechanisms employed by L. orbonalis in resisting insecticide toxicity through accelerated excretion mode. These ABCD and ABCG family genes could be candidate targets in developing genome-assisted pest management strategies in the future.

Efficacy of a RADA-16 peptide hydrogel versus chitosan-based polymer in improving patient comfort during postoperative debridement: A randomized controlled trial.

BACKGROUND: Bioresorbable nasal packing is associated with a decreased incidence of adhesions and bleeding postoperatively after endoscopic sinus surgery (ESS). However, discomfort during postoperative debridement is still a major area of concern for patients. Our objective was to compare the efficacy of a peptide hydrogel to that of a chitosan-based polymer in reducing pain during debridement after ESS. METHODS: A prospective, multicenter, randomized, blinded trial was conducted in adults undergoing bilateral total ethmoidectomy for chronic rhinosinusitis. Participants served as their own controls with each subject receiving the hydrogel in a randomized ethmoid cavity and chitosan-based polymer in the contralateral ethmoid cavity. Participants were evaluated at 1, 4, and 12 weeks postoperatively. Pain during debridement as well as endoscopic evaluation of mucosal healing and hemostasis were measured. RESULTS: Thirty patients who underwent ESS were included in this trial. During the week 1 postoperative debridement, patients reported significantly less pain on the hydrogel-treated side compared to the chitosan-based polymer-treated side. There were no significant differences in bleeding severity, Lund-Kennedy scores, debridement time, or need for further intervention between the two groups. CONCLUSION: This study demonstrated the efficacy of a peptide hydrogel in minimizing pain during postoperative debridement.

Comparative analysis of CBNAAT (GeneXpert) and Ziehl-Nielsen staining test as diagnostic modalities of tuberculosis.

Background: As centuries and millennia passed, tuberculosis remains the major health threat for millions of people. Due to this resurgence of tuberculosis, scientists around the globe have intensified their research efforts to curb down the transmission of this disease and that is the reason which led to the development of new and efficient diagnostic tools, one such being the rapid cartridge-based nucleic acid amplification test which can give the real-time results and that too in a short period of time. These diagnostic tools which are based on the latest technologies help in paving the way to eliminate tuberculosis by early detection method and further prevent the transmission. Aim: In this study, we did a comparative analysis of the effectiveness of CBNAAT technique in the early detection of Mycobacterium tuberculosis over the conventional method of Ziehl-Nielsen microscopy as diagnostic modalities. Materials and Methods: Comparative analysis of CBNAAT (GeneXpert) and Ziehl-Nielsen staining test was done as diagnostic modalities using standard protocols. Results: Out of total 300 samples, 153 were female, 146 were male, while one was transgender. For all these samples, the positivity rates were 48% for the CBNAAT technique and 21.3% for ZN staining method, respectively. For extrapulmonary samples, positivity rates were 49.1% and 11.86% for CBNAAT and ZN microscopy, respectively, while the same for pulmonary samples was 47.7% and 23.6%, respectively. Conclusions: ZN microscopy and CBNAAT are both initial tests for the detection of Mycobacterium tuberculosis, but the introduction of CBNAAT is a boon for peripheral sites. The higher positivity rate of CBNAAT over ZN staining is the icing on the cake with the ability to detect even those cases that could be missed by the latter.

IL-13 and IL-13-induced periostin levels are specifically decreased in patients following endoscopic sinus surgery for chronic rhinosinusitis.

BACKGROUND: Type 2 (T2) inflammation plays a pathogenic role in chronic rhinosinusitis (CRS). The effects of endoscopic sinus surgery (ESS) on T2 inflammation are unknown. OBJECTIVE: The aim of this study was to compare T2 inflammatory biomarkers from middle meatal (MM) mucus for distinguishing patients with CRS from CRS-free patients, identifying major phenotypes (CRS without nasal polyps [CRSsNP] and CRS with nasal polyps [CRSwNP]), assessing endotypic change, and establishing cross-sectional and longitudinal outcomes in patients undergoing ESS. METHODS: MM mucus samples were collected from patients with CRSsNP and patients with CRSwNP before and 6 to 12 months after ESS and compared with samples from CRS-free control patients. T2 biomarkers were evaluated both continuously and using threshold-based definitions of T2 endotype to identify relationships with patient-reported (based on the 22-Item Sinonasal Outcomes Test and Chronic Rhinosinusitis Patient-Reported Outcomes Measure) and clinician-reported (radiographic and endoscopic) severity. Linear mixed models were developed to analyze clinical variables associated with T2 biomarker levels. RESULTS: A total of 154 patients with CRS (89 with CRSsNP and 65 with CRSwNP) were enrolled, with a mean interval of 9 months between ESS and follow-up. An analysis of pre-ESS MM mucus samples revealed elevated levels of T2 mediators in patients with CRSwNP versus in patients with CRSsNP and CRS-free controls. Temporally stable correlations between levels of IL-13 and IL-5, levels of periostin and complement 5a, and levels of eosinophil cationic protein (ECP) and eotaxin-3 were observed. On this basis and on the basis of pathologic significance, levels of IL-13, periostin and ECP were further analyzed. After ESS, levels of IL-13 and periostin decreased significantly, whereas ECP levels remained unchanged. Across pre- and post-ESS evaluation, the T2 endotype was associated with radiographic severity but did not predict outcomes. CRSwNP status and African American race were associated with higher levels of IL-13 and periostin, whereas ECP level was higher in patients undergoing extensive surgery. CONCLUSION: ESS decreased levels of IL-13 and periostin in the middle meatus. T2 inflammation after ESS was correlated with patient- and clinician-reported severity across phenotypes. Pre-ESS T2 inflammation did not predict post-ESS outcomes.

Single-cell transcriptomic analysis of corneal organoids during development.

Corneal organoids are useful tools for disease modeling and tissue transplantation; however, they have not yet been well studied during maturation. We characterized human iPSC-derived corneal organoids at 1, 2, 3, and 4 months of development using single-cell RNA sequencing to determine the cellular heterogeneity at each stage. We found pluripotent cell clusters committed to epithelial cell lineage at 1 month; early corneal epithelial, endothelial, and stromal cell markers at 2 months; keratocytes as the largest cell population at 3 months; and a large epithelial cell population at 4 months. We compared organoid to fetal corneal development at different stages and found that 4-month organoids closely resemble the corneal cellular complexity of the fetal (16 post conception week) and adult cornea. Using RNA velocity trajectory analysis, we found that less differentiated cells appear to give rise to corneal epithelial cells during development.

Lignocellulolytic extremozymes and their biotechnological applications.

Lignocellulolytic enzymes are used in different industrial and environmental processes. The rigorous operating circumstances of these industries, however, might prevent these enzymes from performing as intended. On the other side, extremozymes are enzymes produced by extremophiles that can function in extremely acidic or basic; hot or cold; under high or low salinity conditions. These severe conditions might denature the normal enzymes that are produced by mesophilic microorganisms. The increased stability of these enzymes has been contributed to a number of conformational modifications in their structures. These modifications may result from a few amino acid substitutions, an improved hydrophobic core, the existence of extra ion pairs and salt bridges, an increase in compactness, or an increase in positively charged amino acids. These enzymes are the best option for industrial and bioremediation activities that must be carried out under difficult conditions due to their improved stability. The review, therefore, discusses lignocellulolytic extremozymes, their structure and mechanisms along with industrial and biotechnological applications.

2-{N-[(2,4,5-trichlorophenoxy) acetyl]-N-methylamino}-3-pyrrolidinepropanamide analogs as potential antagonists of Urotensin II receptor.

THE PURPOSE OF THE ARTICLE: To identify novel small molecule antagonists of Urotensin II receptor with acceptable pharmacological profile. MATERIALS AND METHODS: Structure-activity-relationship (SAR) studies on 2-{N-[(2,4,5-trichlorophenoxy) acetyl]-N-methylamino}-3-pyrrolidinepropanamide series were conducted and shortlisted compounds were synthesized and evaluated in in vitro cell-based assays. Human and mouse Urotensin II receptor overexpressing CHO cells were used for calcium release and radioligand binding assays. Initial molecules in this series had solubility and inter-species variability issue in the calcium release assay. We, therefore, conducted SAR to overcome these 2 issues and molecules with accepted in vitro profile were evaluated further in mouse pressor response model to generate the in vivo proof of concept for UII receptor antagonization. RESULTS AND CONCLUSIONS: We report herewith identification of 2-{N-[(2,4,5-trichlorophenoxy)acetyl]-N-methylamino}-3-pyrrolidinepropanamides series to obtain novel small molecule antagonists of Urotensin II receptor with acceptable pharmacological profile.

Mucoid Staphylococcus haemolyticus: an unheeded multidrug-resistant pathogen.

Coagulase-negative Staphylococci (CoNS) are among the most abundant members of human skin microbiome. CoNS have lately been recognized as substantial agents in plethora of infections, especially nosocomial infections in preterm infants and immunocompromised patients. Staphylococcus haemolyticus is the second most common species isolated from blood, and identification is further hindered when there is a deviation in morphology from the classical one. Here, we report an uncommon case of multidrug resistant mucoid S. hemolyticus isolated from blood in a patient of polytrauma. The patient was managed with ceftriaxone-sulbactam, gentamicin, and meropenem as empirical therapy, which was subsequently changed to intravenous vancomycin. The patient showed favorable response to treatment. Mucoid isolates are known to be more virulent and multi-drug resistant than the classical morphotypes. We also conducted systematic review to decipher the prevalence of mucoid S. hemolyticus and linezolid (LZD) resistance in the same. This case highlights the significance of awareness of mucoid phenotypes of Gram-positive cocci for clinical microbiologists to reach accurate identification. Resistance to LZD further underscores the need of restriction policies in hospitals and to roll out antimicrobial stewardship program stringently, so that the growing resistance could be contained.

Sympathetic Ophthalmia - An Overview.

INTRODUCTION: Sympathetic ophthalmia (SO) is rare, bilateral granulomatous panuveitis that typically occurs following penetrating or perforating ocular trauma or surgery. This review aims to provide an update on the etiopathogenesis, clinical presentations, diagnosis and treatment of SO. METHODS: Reports cited in MEDLINE database, that analyzed SO in at least 5 patients, published prior to December 1st, 2021 were included. RESULTS: Initially, SO was associated with penetrating ocular trauma, however, various studies reported an increased incidence of SO after surgical procedures including vitreoretinal surgeries. Multimodal imaging including fluorescein and indocyanine green angiography, optical coherence tomography (OCT) and OCT angiography have added further insights into the understanding of SO. While pulse dose corticosteroids & immunosuppressive drugs are still the treatment of choice, TNF-α blockers & other biologic drugs represent new promising agents. CONCLUSION: There is a growing pool of evidence in understanding the pathogenesis of SO. Novel treatment options have provided better prognosis for this potentially blinding condition.

A rare case of Panophthalmitis due to cobra bite.

PURPOSE: Ocular manifestations of snake bite are rare, ranging from direct injury to the eye from snake venom or indirect injury due to antivenom. We report a rare case of cobra bite causing panophthalmitis due to indirect injury as a result of snake venom toxin related tissue necrosis and susceptibility to secondary infections. METHODS: Observational case report. External photographs, slit lamp photos, ultrasonography of the eye and histopathology of the eviscerated eye were used to characterize and describe the clinical findings. Thirty-nine-years-old male farmer presented with history of cobra snake bite on his right index finger and developed right eye sudden onset pain and redness 3 days later. On examination, features were suggestive of panophthalmitis and the eye had to be eviscerated with scleral excision. CONCLUSION: It is important for ophthalmologist to be aware of such grave consequences of snake bite to be prepared for the emergency management of such cases.

Validation of Hematological Markers in Early Onset Neonatal Sepsis.

Background Neonatal sepsis is considered a ubiquitous worldwide cause of mortality and morbidity in newborn infants. The incidence is 10-50 per 1000 live births. Neutrophil to lymphocyte ratio (NLR) is an easily accessible and cost-effective hematological marker for prompt diagnosis of neonatal sepsis. Aim and objectives The purpose of this study was to analyze the clinical significance of NLR in neonates clinically diagnosed with sepsis and its impact on the management. Methods This retrospective study was conducted on 265 neonates diagnosed with sepsis and compared with 341 healthy controls. The statistical analysis was performed by using the Student's t-test to compare the variables. Result Median NLR levels were significantly higher in patients than in controls. NLR had a modest power of predicting neonatal sepsis, as suggested by an area under a curve of 0.569. Conclusion NLR is an important predictor of neonatal sepsis. There is a significant modest positive correlation between NLR and sepsis.

RETINAL NONPERFUSION IN THE EARLY TREATMENT DIABETIC RETINOPATHY STUDY SEVEN FIELDS COMPARED WITH WIDEFIELD FLUORESCEIN ANGIOGRAPHY: Correlation and Use of Extrapolation Factor.

BACKGROUND: In previous landmark studies on central retinal vein occlusion, retinal nonperfusion assessments were obtained using 7-field (7F) angiography. The widespread current use of widefield imaging allows better visualization of the peripheral retina and more comprehensive estimation of the total area of nonperfusion. The relationship between nonperfusion measurement of 7F and widefield angiography (WFA) in central retinal vein occlusion has not been studied. We aim to identify the correlation of retinal nonperfusion measured within the 7F and on WFA in eyes with central retinal vein occlusion. METHODS: Retinal nonperfusion in participants with central retinal vein occlusion was determined using a 7F Early Treatment Diabetic Retinopathy Study template and the concentric rings method. RESULTS: A total of 153 eyes were included. Pearson correlation test showed a near-perfect positive, linear correlation between the nonperfusion found in the 7F and total retinal nonperfusion on WFA (0.985 95% CI [0.793, 0.999]) The regression line equation for nonperfusion on 7F and WFA was y = 37 + 3.2x. Eyes with 0 disk areas (DA), >0 DA to 10 DA and >10 DA of nonperfusion on 7-fields had on average 23 DA 95% CI (19.20, 27.06), 45 DA 95% CI (35.75, 55.18), and 115 DA 95% CI (88.89, 142.05) on widefield respectively. CONCLUSION: There is a positive and linear relationship between nonperfusion measured by 7F and WFA in central retinal vein occlusion with more than 3-times the amount of nonperfusion identified on WFA. Despite <10 DA no areas of nonperfusion on 7F, there is typically at least 35 DA of nonperfusion on WFA whereas eyes with >10 DA of nonperfusion on 7F had at least 88 DA on WFA.

Immunohistochemical expression of paxillin in potentially malignant disorders and squamous cell carcinoma patients.

Background: Cell adhesion molecules are essential to maintain the integrity of stratified squamous epithelium but their expression has to be dynamic to aid the mobility and turnover of cells. Paxillin is one such multi-domain protein which integrates numerous signals from cell surface receptors, integrins and growth factors. It thus functions as a regulator of various physiological and pathological processes including tissue remodeling, cell motility, gene expression, matrix organization, cell proliferation, metastasis and survival. Hence, the assessment of paxillin expression in normal control, potentially malignant disorders and oral squamous cell carcinoma patients was carried out. Material and Methods: The present retrospective study comprised of 20 each clinically and histologically confirmed case of normal control, potentially malignant disorders, and oral squamous cell carcinomas. All the slides were stained immunohistochemically using Paxillin antibody. Results: The localization, staining intensity and percentage of positivity for paxillin expression was statistically significant among normal control and potentially malignant disorders, whereas oral squamous cell carcinoma showed a non-significant difference. Upon comparison of histopathological grading of potentially malignant disorders, mild versus severe and moderate versus severe epithelial dysplasia showed a statistical significant difference among all the parameters of paxillin expression. However, WDSCC & MDSCC a statistically significant difference among localization and staining intensity of paxillin. Conclusion: Paxillin may play an important role in pathogenesis of oral squamous cell carcinoma by altering the adhesive properties of the tumor cells interacting with the extracellular matrix which in turn affects their invasive behavior and histologic differentiation.

Reply to Comments on Manuscript "Role of Ultra-widefield Imaging in Eales' Disease: A Case Series".

In a comment on our article ''Role of Ultra-widefield Imaging in Eales' Disease: A Case Series", the author asserts that Widefield fluorescein angiography (FA) detected significantly more number of active vasculitis as compared to Optos in our study which is not in line with our conclusion. We agree that Widefield FA is the gold standard for detecting activity in uveitic eye, and in fact, we have mentioned that 'FA was considered the gold standard for diagnosing active vasculitis and clinical examination and Optos findings were compared to FA' in our Materials and methods section.

Treatment outcomes of combination of anti-vascular endothelial growth factor injection and laser photocoagulation in Type 1 ROP and APROP.

PURPOSE: To study treatment outcomes of combination treatment of intravitreal anti-vascular endothelial growth factor (VEGF) injection and laser photocoagulation in Type 1 Retinopathy of Prematurity (ROP) and Aggressive Posterior ROP (APROP). METHODS: This is a retrospective observational study of 87 eyes of 48 premature babies who presented with Type I ROP or APROP and were treated with combination of laser and anti-VEGF therapy. Retrospective evaluation of case records was done to collect data on gestational age, birth weight, age at intervention, anterior segment and fundus findings, intervention with laser and anti-VEGF and response to treatment. Outcome measure was defined as attached retina at posterior pole at last follow-up. RESULTS: Mean gestational age was 29.1 weeks, and mean birth weight was 1226.9 gms. Sixty-six (75.8%) eyes had Type I ROP and 21 (24.1%) eyes had APROP at presentation. Five eyes (3 patients) were lost to follow-up after treatment. Of 82 eyes, 80.5% (66 eyes) showed regression of ROP following combination treatment and 19.5% (16 eyes) needed surgery. Of these, 15 underwent surgery and 12 had successful outcome. Mean follow-up duration of patients was 52 weeks. Finally, 95.1% (78 eyes) had attached retina at posterior pole and 4.9% (4 eyes) had detached retina. CONCLUSION: We conclude that combination therapy is an effective and safe treatment strategy for Type I ROP and APROP.