Highly sensitive and quantitative HiBiT-tagged Nipah virus-like particles: A platform for rapid antibody neutralization studies.

Biocontainment regulations restrict the research on NiV to BSL-4 laboratories, thus limiting the mechanistic studies related to viral entry and allied pathogenesis. Understanding the precise process of viral-particle production and host cell entry is critical for designing targeted therapies or particle-based vaccines. In this study, we have synthesized HiBiT-tagged-NiV-VLPs to ease in-vitro BSL-2 particle handling. We propose a simple yet effective approach of generating substantial amount of HiBiT-tagged NiV-VLPs in vitro by co-expressing viral structural proteins in HEK293T cells. Though homologous to parent virus, the incapacitated replication potential facilitates a BSL-2 handling of these particles. The inclusion of a highly sensitive HiBiT tag on these VLPs allows for a quick detection of viral binding and entry, as well as in assessing the efficiency of neutralizing antibodies in vitro using the NanoBiT technology. The HiBiT-tag binds in high affinity with LgBiT (Large BiT an 18 kDa fusion protein and complementary subunit of HiBiT peptide), and the resultant complex elicits high intensity luminescence in the presence of substrate. The VLPs produced were morphologically and functionally identical to the native virus, and the HiBiT-tag permitted their quick application in viral binding, entry, and antibody neutralization assays. "Thus, we report a simple setting for generating HiBiT-NiV VLPs which can be utilized in a BSL-2 laboratory, to concurrently quantify features of NiV assembly, binding and entry. This also offers an alternate-safe and effective platform for viral based antibody neutralization assays in vitro".

Intraluminal Suture Placement for Management of Hypotony After Glaucoma Drainage Device Surgery.

PRCIS: Intraluminal 3-0 Supramid stent placement can be an effective long-term solution for hypotony after glaucoma drainage device surgery. This procedure may obviate the need for conjunctival dissection. In some cases, additional procedures are required. PURPOSE: To describe the utility of implantation of a multifilament polyamide suture (Supramid) in the tube lumen to reverse hypotony after glaucoma drainage device (GDD) surgery. PATIENTS: Patients who underwent tube revision with intraluminal placement of a 3-0 Supramid stent, with or without external ligature, to manage hypotony following GDD surgery between January 2010-October 2020. METHODS: Resolution of hypotony was defined as IOP >5 mmHg and the absence of hypotony-related structural abnormalities. Overall success was the absence of failure criteria (IOP ≤5 mmHg, >21 mmHg, or <20% reduction below baseline IOP after 3 months for 2 consecutive study visits; additional glaucoma surgery; or loss of light perception), with or without adjunctive ocular hypotensive therapy. RESULTS: Nine eyes of 9 patients underwent placement of a 3-0 Supramid stent with resolution of hypotony in all eyes with mean follow-up duration of 33.3 ±24.0 months. Overall success was achieved in 7 of 9 eyes. Four eyes required intervention after stent placement: 1 eye required selective laser trabeculoplasty followed by replacement of the original stent with a shorter 3-0 Supramid suture and external ligature; 1 eye underwent revision for tube exposure; and 2 eyes underwent laser suture lysis to release an external suture ligature. CONCLUSIONS: In cases of hypotony after GDD, particularly when it is desirable to reduce the extent of additional conjunctival dissection, intraluminal placement of a 3-0 Supramid stent via an intracameral approach can be an effective long-term solution. Post-operative adjustments were required in many cases.

Oral health in cancer palliative care: cross-sectional study.

OBJECTIVES: Patients on anticancer therapy attending palliative care services often have oral health problems, but not enough is known in this regard (in India). This cross-sectional study aimed to elucidate this issue. METHODS: Participants were 98 patients with metastatic cancer (52 males, 46 females) who attended the palliative care clinic and were assessed by a multidisciplinary team consisting of dental health professionals working alongside the palliative care team, between August 2021 and October 2022. Their demographic and medical details were collected from the charts. Details about oral health were assessed using the National Cancer Institute Common Terminology Criteria for Adverse Events for dysphagia and xerostomia, Dental Caries Decayed, Missing, and Filled Teeth (DMFT) index, and the WHO Mucositis scale. Quality of life was assessed using the European Organisation for Research and Treatment of Cancer Quality of Life Module for Oral Health (EORTC QLQ-OH15). RESULTS: Patients had an average age of 58 years, with 46.9% having an Eastern Cooperative Oncology Group score of 1, and 61 (64.9%) receiving anticancer treatment along with palliative care. Dental issues were prevalent, with 39.5% classified as DMFT score grade 2. Subgroup analysis showed higher mean DMFT scores in head/neck cancers, particularly in those receiving radiation therapy (2.3) versus without (1.7), and older age (p<0.05). CONCLUSION: Patients with cancer on anticancer treatment attending a palliative care clinic, especially those with head/neck cancers and older age, had poor oral health. Further prospective research with a dentist on the team is warranted to assess its impact.

Safety protocols at the American Board of Pediatric Dentistry Oral Clinical Examination during COVID-19.

BACKGROUND: There are continued concerns about holding large group gatherings with the ongoing COVID-19 pandemic. The American Board of Pediatric Dentistry (ABPD) made the decision to cancel the Oral Clinical Examination (OCE) in 2020 with a goal to host the exam in 2021. This cancellation postponed the board certification of over 400 pediatric dentists. There is limited published data on safety protocols for in-person examinations. AIM: To determine if safety protocols utilized in the administration of the 2021 ABPD Oral Clinical Examination (OCE) were reassuring to candidates and examiners, and effective at addressing prevention of COVID-19 infection. DESIGN: An electronic survey was emailed to candidates and examiners following completion of the board examination process. The candidate survey had 784 and the examiner survey had 108 eligible participants. Eight questions overlapped between the two surveys. Descriptive statistics were utilized. p-values < 0.05 were considered significant. RESULTS: Three hundred ninety-four candidates (response rate = 50.3%) and 97 examiners (response rate = 89.8%) completed their respective surveys. The majority of candidates (86.8%) and examiners (94.8%) felt safe participating the OCE, and found COVID-19 protocols reassuring, 69.3% and 89.7%. CONCLUSIONS: Participants agreed that safety protocols enacted by the ABPD kept everyone safe and led to a successful and safely conducted high-stakes oral clinical examination during the COVID-19 pandemic.

Inactivated whole-virion vaccine BBV152/Covaxin elicits robust cellular immune memory to SARS-CoV-2 and variants of concern.

BBV152 is a whole-virion inactivated vaccine based on the Asp614Gly variant. BBV152 is the first alum-imidazoquinolin-adjuvanted vaccine authorized for use in large populations. Here we characterized the magnitude, quality and persistence of cellular and humoral memory responses up to 6 months post vaccination. We report that the magnitude of vaccine-induced spike and nucleoprotein antibodies was comparable with that produced after infection. Receptor binding domain-specific antibodies declined against variants in the order of Alpha (B.1.1.7; 3-fold), Delta (B.1.617.2; 7-fold) and Beta (B.1.351; 10-fold). However, pseudovirus neutralizing antibodies declined up to 2-fold against the Delta followed by the Beta variant (1.7-fold). Vaccine-induced memory B cells were also affected by the Delta and Beta variants. The SARS-CoV-2-specific multicytokine-expressing CD4+ T cells were found in ~85% of vaccinated individuals. Only a ~1.3-fold reduction in efficacy was observed in CD4+ T cells against the Beta variant. We found that antigen-specific CD4+ T cells were present in the central memory compartment and persisted for at least up to 6 months post vaccination. Vaccine-induced CD8+ T cells were detected in ~50% of individuals. Importantly, the vaccine was capable of inducing follicular T helper cells that exhibited B-cell help potential. These findings show that inactivated vaccine BBV152 induces robust immune memory to SARS-CoV-2 and variants of concern that persists for at least 6 months after vaccination.

Inactivated virus vaccine BBV152/Covaxin elicits robust cellular immune memory to SARS-CoV-2 and variants of concern

The characteristics of immune memory established in response to inactivated SARS-CoV-2 vaccines remains unclear. We determined the magnitude, quality and persistence of cellular and humoral memory responses up to 6 months after vaccination with BBV152/Covaxin. Here, we show that the quantity of vaccine-induced spike- and nucleoprotein-antibodies is comparable to that following natural infection and the antibodies are detectable up to 6 months. The RBD-specific antibodies decline in the range of 3 to 10-fold against the SARS-CoV-2 variants in the order of alpha (B.1.1.7) > delta (B.1.617.2) > beta (B.1.351), with no observed impact of gamma (P.1) and kappa (B.1.617.1) variant. We found that the vaccine induces memory B cells, similar to natural infection, which are impacted by virus variants in the same order as antibodies. The vaccine further induced antigen-specific functionally potent multi-cytokine expressing CD4+ T cells in [~]85% of the subjects, targeting spike and nucleoprotein of SARS-CoV-2. Marginal [~]1.3 fold-reduction was observed in vaccine-induced CD4+ T cells against the beta variant, with no significant impact of the alpha and the delta variants. The antigen-specific CD4+ T cells were populated in the central memory compartment and persisted up to 6 months of vaccination. Importantly the vaccine generated Tfh cells that are endowed with B cell help potential, similar to the Tfh cells induced after natural infection. Altogether, these findings establish that the inactivated virus vaccine BBV152 induces robust immune memory to SARS-CoV-2 and variants of concern, which persist for at least 6 months after vaccination. This study provides insight into the attributes of BBV152-elicited immune memory, and has implication for future vaccine development, guidance for use of inactivated virus vaccine, and booster immunization.

Dental Topics for Plastic Surgeons, Part Three: Infection and Trauma of the Dentition.

ABSTRACT: Conditions that affect dental and periodontal structures receive sparse coverage in the plastic surgery literature, yet a working knowledge of this subject matter is important in certain areas of clinical practice and a fundamental understanding is part of plastic surgery competency tested on the in-service and written board examinations. This 4-part series written to provide plastic surgeons with a working knowledge of dental topics that may be relevant to their clinical practice. This section, Part III, covers inflammatory and infectious conditions of the dentition and related structures, as well as dentoalveolar trauma.

Synthesis, characterization, in silico, and in vitro biological screening of coordination compounds with 1,2,4-triazine based biocompatible ligands and selected 3d-metal ions.

A bidentate Schiff base ligand, MHMMT, obtained from 1,2,4-triazine derivative and 4-hydroxy-3-methoxy benzaldehyde and its Fe(III), Co(II), Ni(II), Cu(II), and Zn(II) complexes were synthesised in ethanolic media and characterized by various analytical techniques like elemental analyses, magnetic susceptibility measurements, FTIR, UV-VIS, proton NMR, ESR, spectroscopic and thermogravimetric studies. Various geometries like a tetrahedral for Co(II) and Zn(II) complexes, an octahedral for Fe(III) and Ni(II) complexes, and square planar for Cu(II) complex has been assigned. For all metals complexes except Co(II), a 2:1 ligand to metal ratio is observed, while Co(II) complex has a 1:1 ratio. In accordance with the probable activity spectra of substances as obtained from PASS analysis, in vitro α-amylase inhibition studies by starch-iodine method for ligand and complexes except that of Fe(III) and anticancer screening against human breast cancer cell lines MCF-7 using MTT assay for Fe(III) complex were conducted. The tested compounds were found to be good α-amylase inhibitors, characteristically similar to most of the antidiabetic drugs. Among the compounds, Cu(II) complex exhibited the highest α-amylase inhibitory activity. Furthermore, ligand and complexes were also exposed to in vitro antimicrobial activities, drug-likeness, bioactivity score prediction by Molinspiration software. Molecular docking analysis of selected compounds on α-amylase and VEGFR-2 kinase were carried out for confirming the experimental observations.

Cytokine Storm in COVID-19-Immunopathological Mechanisms, Clinical Considerations, and Therapeutic Approaches: The REPROGRAM Consortium Position Paper.

Cytokine storm is an acute hyperinflammatory response that may be responsible for critical illness in many conditions including viral infections, cancer, sepsis, and multi-organ failure. The phenomenon has been implicated in critically ill patients infected with SARS-CoV-2, the novel coronavirus implicated in COVID-19. Critically ill COVID-19 patients experiencing cytokine storm are believed to have a worse prognosis and increased fatality rate. In SARS-CoV-2 infected patients, cytokine storm appears important to the pathogenesis of several severe manifestations of COVID-19: acute respiratory distress syndrome, thromboembolic diseases such as acute ischemic strokes caused by large vessel occlusion and myocardial infarction, encephalitis, acute kidney injury, and vasculitis (Kawasaki-like syndrome in children and renal vasculitis in adult). Understanding the pathogenesis of cytokine storm will help unravel not only risk factors for the condition but also therapeutic strategies to modulate the immune response and deliver improved outcomes in COVID-19 patients at high risk for severe disease. In this article, we present an overview of the cytokine storm and its implications in COVID-19 settings and identify potential pathways or biomarkers that could be targeted for therapy. Leveraging expert opinion, emerging evidence, and a case-based approach, this position paper provides critical insights on cytokine storm from both a prognostic and therapeutic standpoint.

Key Strategies for Clinical Management and Improvement of Healthcare Services for Cardiovascular Disease and Diabetes Patients in the Coronavirus (COVID-19) Settings: Recommendations From the REPROGRAM Consortium.

Patients with cardiovascular disease and diabetes are at potentially higher risk of infection and fatality due to COVID-19. Given the social and economic costs associated with disability due to these conditions, it is imperative that specific considerations for clinical management of these patients be observed. Moreover, the reorganization of health services around the pandemic response further exacerbates the growing crisis around limited access, treatment compliance, acute medical needs, and mental health of patients in this specific subgroup. Existing recommendations and guidelines emanating from respective bodies have addressed some of the pressure points; however, there are variations and limitations vis a vis patient with multiple comorbidities such as obesity. This article will pull together a comprehensive assessment of the association of cardiovascular disease, diabetes, obesity and COVID-19, its impact on the health systems and how best health systems can respond to mitigate current challenges and future needs. We anticipate that in the context of this pandemic, the cardiovascular disease and diabetes patients need a targeted strategy to ensure the harm to this group does not translate to huge costs to society and to the economy. Finally, we propose a triage and management protocol for patients with cardiovascular disease and diabetes in the COVID-19 settings to minimize harm to patients, health systems and healthcare workers alike.

Dental Topics for Plastic Surgeons, Part Two: Abnormal Tooth Development, Conditions, and Treatment.

Conditions that affect dental and periodontal structures receive sparse coverage in the plastic surgery literature, yet a working knowledge of this subject matter is important in certain areas of clinical practice and a fundamental understanding is part of plastic surgery competency tested on the in-service and written board examinations. This four-part series written to provide plastic surgeons with a working knowledge of dental topics that may be relevant to their clinical practice. This section, Part II, covers abnormal tooth development and related conditions.

Dental Topics for Plastic Surgeons, Part One: Normal Anatomy, Growth and Development.

Normal and abnormal conditions affecting the dentition and the periodontal structures receive sparse coverage in the plastic surgery literature, textbooks, and training programs. Nevertheless, a working knowledge of this subject matter is important in certain areas of clinical practice, and a fundamental understanding is often part of plastic surgery competency tested for in the In-service and written board examinations. This four-part series is written to provide plastic surgeons with a working knowledge of relevant dental topics. Part 1 covers fundamental aspects of normal dental embryology, growth and anatomy.

Suprachoroidal hemorrhage after micropulse cyclophotocoagulation diode therapy.

PURPOSE: To describe the diagnosis and management of a patient with primary open angle glaucoma (POAG) who developed suprachoroidal hemorrhage (SCH) after micropulse cyclophotocoagulation (MPCPC) therapy. OBSERVATIONS: A 77 year-old Caucasian man with end-stage POAG and multiple medical comorbidities including coronary artery disease on anticoagulation presented with 2 days of episodic, severe left eye pain 2 weeks after undergoing MPCPC diode in the left eye. His visual acuity was count fingers at 2 feet and his intraocular pressure (IOP) was 44. He had a shallow anterior chamber that was open to trabecular meshwork on gonioscopy. His dilated fundus exam showed large, dome-shaped hemorrhagic choroidals, which were confirmed on ultrasound. He was medically managed with analgesics, cycloplegics, topical steroids and IOP lowering medications. He was closely followed with serial b-scans and the SCH decreased in size without surgical intervention, however, his visual acuity did not improve from presentation. CONCLUSIONS AND IMPORTANCE: MPCPC diode has been increasingly used in refractory glaucoma and is considered to be a relatively safe procedure. Suprachoroidal hemorrhage has not yet been reported after MPCPC diode. This case demonstrates how devastating complications such as SCH can still occur with lower energy CPC therapy especially in the setting of post-procedural hypotony, and emphasizes the importance of prevention especially in high-risk patients.

Parafoveal vessel changes in primary open-angle glaucoma and normal-tension glaucoma using optical coherence tomography angiography.

PURPOSE: To evaluate parafoveal and peripapillary perfusion in healthy, glaucoma suspect, normal-tension glaucoma, and primary open-angle glaucoma subjects. PATIENTS AND METHODS: This was a retrospective cross-sectional study with optical coherence tomography angiography imaging with RTVue XR Avanti (Optovue, Inc., Fremont, CA) of 56 eyes (14 healthy, 14 glaucoma suspect, 16 normal-tension glaucoma, and 12 primary open-angle glaucoma) at a tertiary academic referral center. Parafoveal and peripapillary superficial vessel density and parafoveal superficial retinal thickness were the main parameters of interest. Area under receiver operating characteristic curves were calculated. RESULTS: There were significant decreases in parafoveal superficial vessel density in primary open-angle (40.06±4.54%, P<0.001) and normal-tension glaucoma (42.82±5.16%, P=0.010) but not suspect eyes (45.72±4.37%, P=0.916) compared to healthy eyes (48.10±2.82%). Similarly, decreases were observed in parafoveal inner retinal thickness in primary open-angle (83.19±14.29 µm, P<0.001) and normal-tension glaucoma eyes (94.97±12.44 µm, P=0.035), but not suspect eyes (99.93±9.00 µm, P=0.648), compared to healthy controls (107.00±9.55 µm). Only primary open-angle glaucoma eyes displayed significant changes in peripapillary vessel density (37.63±7.19%) compared to healthy controls (49.12±2.80%, P<0.001). Further statistical adjustment for sex and age revealed a significant decrease in parafoveal vessel density in suspects relative to controls (P=0.039). Diagnostic accuracy of parafoveal vessel density was high with an area under the curve of 0.833±0.073 for normal-tension glaucoma and 0.946±0.049 for primary open-angle glaucoma. CONCLUSION: Parafoveal vessel density was significantly reduced in glaucomatous eyes, with good diagnostic accuracy. These findings provide further evidence that these changes may be useful in the diagnosis and monitoring of disease in glaucoma patients.

Highly selective inhibition of α-glucosidase by green synthesised ZnO nanoparticles - In-vitro screening and in-silico docking studies.

Diabetic control through the inhibition of carbohydrate hydrolysing enzymes is established as an effective strategy. Many of the inorganic materials have already been investigated as enzyme inhibitors. Present study investigates the in-vitro antidiabetic activity of ZnO nanoparticles assessing their inhibition efficiency on α-glucosidase and α-amylase. The nanoparticles obtained with average dimeson of 11, 32 and 49 nm via green strategy were subjected to in-vitro antidiabetic assays. The samples were particularly selective for α-glucosidase while very mild inhibitors for α-amylase. ZnO sample with particle dimension of 32 nm was found to be the most potent inhibitor capable of preventing 97.86% enzymatic action. The IC50, and CC50 values of the sample were determined as 1.24 µg/mL and 88.89 µg/mL respectively. The selectivity index (SI) value obtained is 71.68 which indicates good selectivity towards enzymes inhibition rather than the host body. Molecular docking models were generated for ZnO association with α-glucosidase and possible binding sites were recognized.

N-Methyl-D-Aspartate Encephalitis our Experience with Diagnostic Dilemmas, Clinical Features, and Outcome.

INTRODUCTION: A neuropsychiatric syndrome characterized by a wide spectrum of clinical manifestations. It is seen in patients with antibodies against NR1-NR2 heteromers of the NMDA receptor. As the spectrum is mainly psychiatric most patients are treated as psychiatric disease resulting in huge diagnostic delay. PATIENT AND METHODS: Here we describe 29 patients with NMDA encephalitis seen by the authors in the last five years. Percentage of Transfected cells showing granular cytoplasmic florescence was considered for positivity and severity both in CSF and serum. Their presenting diagnosis, clinical features and the dilemmas, alarming gaps, laboratory data, response to treatment and relapses are discussed. OBSERVATIONS: All patients presented with a spectrum of psychiatric symptoms varying from panic to severe aggression, seizures, chorea, hemiplegia, catatonia, mitgehen, mutism, delirium, mania and memory problems. EEG is invariably abnormal as against imaging. CONCLUSION: NMDA receptor mediated encephalitis should be suspected in all children and females of adolescent age with refractory neuropsychiatric syndrome. Both CSF and serum should be tested and regular follow up for relapses and neoplasms is mandatory.

Factors Determining the Clinical Spectrum, Course and Response to Treatment, and Complications in Seronegative Patients with Central Nervous System Tuberculosis.

INTRODUCTION: Tuberculous meningitis remains a major health issue in the community affecting young adults of both genders predominantly from rural background. In India, the disease continues to kill 2 people every 3 min or nearly 1000 daily, according to the Tuberculosis Control Society India. Tuberculosis (TB) of central nervous system (CNS) is the most devastating form of TB. As this disease is associated with very high prevalence in young adults and will ultimately contribute to great workforce loss, we decided to assess the factors deciding the disease and its course in our patients. PATIENTS AND METHODS: Seronegative patients with probable CNS TB and attending our outpatient department were included and followed up for 2 years. RESULTS: Low body mass index, low proteins, albumin, and low CD3 and CD4, pulmonary TB appears to be a common denominator in a vast majority of these patients. Delay in diagnosis and hyponatremia contributes to morbidity. The location of exudates causes morbidity when they are seen in optochiasmal region. Bacille Calmette-Guerin (BCG) vaccination status in the community appears to be very small. CONCLUSION: CNS TB causes considerable morbidity and mortality in rural young adults resulting in severe manpower loss. Awareness into the possible role of BCG in reducing the complications of TB, role of nutrition and immunity even in seronegative patients and high-degree suspicion in medical professionals can bring down the burden of this deadly disease in the society.

Right-sided endocarditis: eustachian valve and coronary sinus involvement.

Infective endocarditis in patients with intravenous drug use commonly involves right-sided heart valves. Eustachian valve (EV) endocarditis is not commonly seen given the valve's infrequent presence. Involvement of the coronary sinus (CS) with endocarditis is also an unusual finding. We present a case with echocardiographic findings consistent with EV endocarditis along with CS involvement, which appropriately responded to antibiotics.