Evaluation of Postoperative Pain Perception Incidence after Single-Visit Versus Multiple-Visit Root Canal Therapy: A Randomized Controlled Trial.

Background: Newer concept of root canal therapy is single-visit one rather than conventional multivisit therapy. Major complaint of patients after root canal therapy is mild or severe pain. Aim: This study aims to assess the prevalence of postoperative discomfort after root canal treatment conducted in both single and multiple visits. Materials and Methods: An experiment using a randomized controlled trial design was conducted, including a total of 80 participants. These individuals were then separated into two groups, with each group consisting of 40 participants. Group A had single-visit root canal therapy, whereas Group B received multivisit root canal treatment. The incidence of pain after therapy was evaluated and compared at four time points: 6 hours, 12 hours, 24 hours, and 48 hours after obturation. Results: The level of pain experienced by patients in Group B was notably greater in comparison with individuals in Group A. Nevertheless, there was no statistically significant difference in the level of pain reported by the patients 48 hours after treatment in either of the groups. Conclusion: There is no significant difference in the occurrence of discomfort after endodontic treatment conducted in either a single visit or many visits, as seen during a 48-hour period after obturation.

Prediction and Diagnosis of Breast Cancer Using Machine and Modern Deep Learning Models.

Background &Objective: Carcinoma of the breast is one of the major issues causing death in women, especially in developing countries. Timely prediction, detection, diagnosis, and efficient therapies have become critical to reducing death rates. Increased use of artificial intelligence, machine, and deep learning techniques create more accurate and trustworthy models for predicting and detecting breast cancer. This study aims to examine the effectiveness of several machine and modern deep learning models for prediction and diagnosis of breast cancer. METHODS: This research compares traditional machine learning classification methods to innovative techniques that use deep learning models. Established usual classification models such as k-Nearest Neighbors (kNN), Gradient Boosting, Support Vector Machine (SVM), Neural Network, CN2 rule inducer, Naive Bayes, Stochastic Gradient Descent (SGD), and Tree, and deep learning models such as Neural Decision Forest and Multilayer Perceptron used. The investigation, which was carried out using the Orange and Python tools, evaluates their diagnostic effectiveness in breast cancer detection. The evaluation uses UCI's publicly accessible Wisconsin Diagnostic Data Set, enabling transparency and accessibility in the study approach. RESULT: The mean radius ranges from 6.981 to 28.110, while the mean texture runs from 9.71 to 39.28 in malignant and benign cases. Gradient boosting and CN2 rule inducer classifiers outperform SVM in accuracy and sensitivity, whereas SVM has the lowest accuracy and sensitivity at 88%. The CN2 rule inducer classifier achieves the greatest ROC curve score for benign and malignant breast cancer datasets, with an AUC score of 0.98%. MLP displays distinguish positive and negative classes, with a higher AUC-ROC of 0.9959. with accuracy of 96.49%, precision of 96.57%, recall of 96.49%, and an F1-Score of 96.50%. CONCLUSION: Among the most commonly used classifier models, CN2 rule and  GB performed better than other models. However, MLP from deep learning produced the greatest overall performance.

To Assess the Efficacy of Various Disinfection and Hemostasis Procedures in Providing Postoperative Pain Relief Following Pulpotomy in Cases of Symptomatic Irreversible Pulpitis.

AIM: The purpose of this study is to assess the efficacy of various disinfection and hemostasis procedures in providing postoperative pain relief following pulpotomy in cases of symptomatic irreversible pulpitis. MATERIALS AND METHODS: The data from a cohort of 50 patients who received treatment with sodium hypochlorite (NaOCl) and another cohort of 50 patients who received treatment with potassium titanyl phosphate (KTP) laser were subjected to analysis. The patients were provided a questionnaire to evaluate pain levels before and after surgery. The patients documented their levels of postoperative pain at specific time intervals, including the sixth hour, first day, second day, third day, and eighth day. This was done using a 100 mm visual analog pain scale, where a marking of 0 mm indicated no pain and a marking of 100 mm indicated the highest level of pain, reflecting the severity of the pain experienced. RESULTS: The pain score of group B was significantly lower than that of group A on the first day. While no notable disparity was detected among the groups during the remaining postoperative periods, it is worth noting that the KTP laser exhibited comparatively lower pain scores. In both groups, the initial pain score before surgery was found to be significantly higher than the pain scores recorded at all subsequent time intervals after surgery. Within group A, it was observed that the pain score during the sixth hour after the surgical procedure was notably greater compared to the pain scores recorded during all other time intervals following the operation. In group B, the pain score at the sixth hour exhibited a statistically significant increase compared to the pain scores observed on the third day and eighth day. CONCLUSION: The KTP laser group exhibited a lower postoperative pain score in comparison to the NaOCl group. The utilization of KTP laser-assisted pulpotomy demonstrated enhanced efficacy in alleviating pain among individuals diagnosed with symptomatic irreversible pulpitis.

DNA Deamination Is Required for Human APOBEC3A-Driven Hepatocellular Carcinoma In Vivo.

Although the APOBEC3 family of single-stranded DNA cytosine deaminases is well-known for its antiviral factors, these enzymes are rapidly gaining attention as prominent sources of mutation in cancer. APOBEC3's signature single-base substitutions, C-to-T and C-to-G in TCA and TCT motifs, are evident in over 70% of human malignancies and dominate the mutational landscape of numerous individual tumors. Recent murine studies have established cause-and-effect relationships, with both human APOBEC3A and APOBEC3B proving capable of promoting tumor formation in vivo. Here, we investigate the molecular mechanism of APOBEC3A-driven tumor development using the murine Fah liver complementation and regeneration system. First, we show that APOBEC3A alone is capable of driving tumor development (without Tp53 knockdown as utilized in prior studies). Second, we show that the catalytic glutamic acid residue of APOBEC3A (E72) is required for tumor formation. Third, we show that an APOBEC3A separation-of-function mutant with compromised DNA deamination activity and wildtype RNA-editing activity is defective in promoting tumor formation. Collectively, these results demonstrate that APOBEC3A is a "master driver" that fuels tumor formation through a DNA deamination-dependent mechanism.

Flow cytometry profiling of cellular immune response in COVID-19 infected, recovered and vaccinated individuals.

INTRODUCTION: SARS-CoV-2 has infected over 753 million individuals and caused more than 6.8 million deaths globally to date. COVID-19 disease severity has been associated with SARS-CoV-2 induced hyper inflammation and the immune correlation with its pathogenesis remains unclear. Acute viral infection is characterised by vigorous coordinated innate and adaptive activation, including an early cellular response that correlates well with the amplitude of virus specific humoral response. OBJECTIVE: The present study covers a wide spectrum of cellular immune response against COVID-19, irrespective of infection and vaccination. METHODS: We analysed immune status of (a) COVID-19 hospitalised patients including deceased and recovered patients, and compared with home isolated and non-infected healthy individuals, and (b) infected home isolated individuals with vaccinated individuals, using flow cytometry. We performed flow cytometry analysis of PBMCs to determine non-specific cell-mediated immune response. RESULTS: The immune response revealed extensive induction and activation of multiple immune lineages, including T and B cells, Th17 regulatory subsets and M1, M2 macrophages in deceased and hospitalised recovered patients, vaccinated and healthy individuals. Compromised immune cell expression was observed in deceased patients even in later stages, while expression was restored in hospitalised recovered patients and home isolated individuals. CONCLUSION: The findings associated with recovery and convalescence define a new signature of cellular immune response that persists in individuals with SARS-CoV-2 infection and vaccination. The findings will help in providing a better understanding of COVID-19 disease and will aid in developing better therapeutic strategies for treatment.

Pharmacological tumor PDL1 depletion with chlorambucil treats ovarian cancer and melanoma: improves antitumor immunity and renders anti-PDL1-resistant tumors anti-PDL1-sensitive through NK cell effects.

BACKGROUND: Tumor intracellular programmed cell death ligand-1 (PDL1) mediates pathologic signals that regulate clinical treatment responses distinctly from surface-expressed PDL1 targeted by αPDL1 immune checkpoint blockade antibodies. METHODS: We performed a drug screen for tumor cell PDL1 depleting drugs that identified Food and Drug Administration (FDA)-approved chlorambucil and also 9-[2-(phosphonomethoxy)ethyl] guanine. We used in vitro and in vivo assays to evaluate treatment and signaling effects of pharmacological tumor PDL1 depletion focused on chlorambucil as FDA approved, alone or plus αPDL1. RESULTS: PDL1-expressing mouse and human ovarian cancer lines and mouse melanoma were more sensitive to chlorambucil-mediated proliferation inhibition in vitro versus corresponding genetically PDL1-depleted lines. Orthotopic peritoneal PDL1-expressing ID8agg ovarian cancer and subcutaneous B16 melanoma tumors were more chlorambucil-sensitive in vivo versus corresponding genetically PDL1-depleted tumors. Chlorambucil enhanced αPDL1 efficacy in tumors otherwise αPDL1-refractory, and improved antitumor immunity and treatment efficacy in a natural killer cell-dependent manner alone and plus αPDL1. Chlorambucil-mediated PDL1 depletion was relatively tumor-cell selective in vivo, and treatment efficacy was preserved in PDL1KO hosts, demonstrating tumor PDL1-specific treatment effects. Chlorambucil induced PDL1-dependent immunogenic tumor cell death which could help explain immune contributions. Chlorambucil-mediated PDL1 reduction mechanisms were tumor cell-type-specific and involved transcriptional or post-translational mechanisms, including promoting PDL1 ubiquitination through the GSK3ß/ß-TRCP pathway. Chlorambucil-mediated tumor cell PDL1 depletion also phenocopied genetic PDL1 depletion in reducing tumor cell mTORC1 activation and tumor initiating cell content, and in augmenting autophagy, suggesting additional treatment potential. CONCLUSIONS: Pharmacological tumor PDL1 depletion with chlorambucil targets tumor-intrinsic PDL1 signaling that mediates treatment resistance, especially in αPDL1-resistant tumors, generates PDL1-dependent tumor immunogenicity and inhibits tumor growth in immune-dependent and independent manners. It could improve treatment efficacy of selected agents in otherwise treatment-refractory, including αPDL1-refractory cancers, and is rapidly clinically translatable.

Effect of various malocclusion on maximal bite force- a systematic review.

Objective: This systematic review evaluates the variation in molar bite force (MBF) with the type and severity of dental malocclusion in young adults with permanent dentition. Methods: We searched seven electronic databases until December 31, 2021, and identified 1898 articles, of which 22 full-texts were reviewed. Eight clinical studies with subjects having permanent dentition with various dental malocclusions and quantifying maximal bite force were included for the review. Newcastle Ottawa scale was used to assess the risk of bias and GRADE to study the certainty of evidence. Articles were evaluated for the primary outcome (variation of MBF in different malocclusion groups) and confounding factors affecting MBF. Results: All studies measured MBF in individuals with normal and malocclusion, with 2329 subjects having permanent dentition. A positive correlation of Class I normal occlusion with the bite force was seen compared to Class II and III malocclusion. Unilateral crossbite patients had lesser bite force. Six studies with 1023 males and 1175 females showed MBF more in males than females. In 3 studies (332 subjects), no significant difference for MBF between the right and left sides of the jaws was measured. Conclusion: MBF decreases significantly with vertical and transverse craniofacial and dental discrepancy. Normal sagittal occlusion has more molar bite force than patients with different malocclusions. Also, MBF is more in males than females, and it increases with age. Registration: PROSPERO CRD42021249328.

Development of robust, indigenous ELISA for detection of IgG antibodies against CoV-2 N and S proteins: mass screening.

The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic has adversely affected humankind and caused millions of deaths globally since January 2020. Robust and quick serological tests such as antibody detection assays for SARS-CoV-2 provide relevant information and aid in the process of vaccine development and diagnostics, as well as in sero-epidemiological monitoring of antibody response to the virus. The receptor-binding domain (RBD) of spike and nucleocapsid protein are specific targets for detecting SARS-CoV-2 antibodies. Here, we present the development of a stable spike (S) and nucleocapsid (N) protein-based ELISA antibody detection test "CoroSuchak," with 99% sensitivity, 98% specificity, cost-effective, and detection in a minimum time for serodiagnosis and mass screening of the population for antibodies against SARS-CoV-2. Blood samples were analyzed from 374 SARS-CoV-2 reverse transcription-polymerase chain reaction (RT-PCR) positive, 772 negative and asymptomatic, and 874 random groups of subjects. We found that the antibody titer was significantly higher (p < 0.0001) in infected and vaccinated group compared to the only vaccinated and only infected group. Using enzyme-linked immunosorbent assay (ELISA), we detected SARS-CoV-2 immunoglobulin G (IgG) antibodies in 118/123 (96%) infected individuals, 570/653 (87%) non-infected but vaccinated individuals, 231/237 (97%) individuals who were both infected and vaccinated, and 499/874 (57%) from randomly selected individuals from the first and second waves of the pandemic. Similarly in the third wave, 14/14 (100%) infected and 16/20 (80%) RT-PCR-negative but symptomatic subjects were detected. Thus, the highly sensitive and specific in-house developed ELISA antibody detection kit "CoroSuchak" is extremely useful to determine the seroprevalence of SARS-CoV-2 antibodies in the coronavirus-exposed population. KEY POINTS: •Indigenous kit using a combination of spike and nucleocapsid proteins and peptide sequences. •High sensitivity and specificity to detect variants. •Highly sensitive for mass screening.

In-situ generation of fluorescent silica nano-aggregates of silatranyl appended furfural Schiff base and its application to the spectrofluorimetric analysis of phenolic brominated flame retardants in aqueous medium.

A silatranyl appended furfural Schiff base (Silt-FUR) has been synthesized and characterized by spectroscopic techniques, elemental analysis and mass spectrometry. The dissolution of Silt-FUR in methanol-water (90:10 v/v) results in the formation of fluorescent nano-aggregates due to the hydrolysis of the silatranyl ring. The formation of nano-aggregates has been confirmed by dynamic light scattering, scanning electron microscopy and transmission electron microscopy. The nano-aggregates exhibit quenching of fluorescence in the presence of phenolic brominated flame retardants such as 3,3',5,5'-tetrabromobisphenol A, 2,4-dibromophenol, 2,4,6-tribromophenol, and pentabromophenol. Density-Functional Theory and NMR titration suggest that acid-base pair formation between azomethinic functionality and flame retardants is the main cause of quenching of fluorescent signal as it causes photoinduced electron transfer. Due to the excellent spectrofluorimetric response of Silt-FUR nano-aggregates to detect brominated phenols, a spectrofluorimetric method has been standardized for the quantification of brominated flame retardants. The detection limit for pentabromophenol obtained is 0.432 µM under optimal experimental conditions, and the linear range of the determination is 0.0495-1.35 µM. Thus, the in-situ generation of nano-aggregates offers a user-friendly method for the detection, quantification and extraction of the brominated phenols with exceptionally high sensitivity and selectivity for pentabromophenol.

Severe heat stress modulated nuclear factor erythroid 2-related factor 2 and macrophage migration inhibitory factor pathway in rat liver.

Heat stress impairs physiology and overall functionality of the body at tissue and organ level in animals. Liver being a vital organ performs more than hundreds regulatory functions of the body. Present study investigates the modulation of molecular pathways that are responsible for liver damage triggered by heat stress. Male Sprague dawley rats were exposed to heat stress (45 °C) in heat simulation chamber till core temperature reaches 40 °C and 42 °C in 25 and 42 min respectively. For in-depth evaluation of liver functions during severe heat stress, hepatic transcriptome and proteome were analysed by microarray and two dimensional gel electrophoresis respectively. Results revealed major alterations in redox status, inflammation, mitochondrial dysfunction and proteostasis related pathways. Data of molecular pathway analysis demonstrate that nuclear factor erythroid 2-related factor 2 (NRF-2) mediated oxidative stress response and macrophage migration inhibitory factor (MIF) regulated inflammatory pathways were upregulated in severe heat stressed liver. Expression levels of downstream molecules of above pathways such as heat shock protein 90AB 1, peroxiredoxin 5, Jun N-terminal kinases 1/2, heme-oxygenase 1, apolipoprotein 1 and interleukin 10 were examined and result suggested the upregulation of these genes modulates the NRF-2 and MIF regulated pathways in heat stressed liver. Irregularity in molecular signalling networks lead to mitochondrial dysfunction indicated by upregulation of ATP synthase ß and peroxiredoxin 1 along with decreased levels of glucose-6-phosphate dehydrogenase and enhanced activity of cytochrome c in liver mitochondria. Thus, current study demonstrated heat induced alterations in key liver functions were regulated by NRF-2 and MIF pathways.

Delaying Intraoral Radiographs during the COVID-19 Pandemic: A Conundrum.

BACKGROUND: The COVID-19 pandemic has made dentists very assiduous about cross-infection during dental treatment, thereby delaying dental radiographs for treatment. However, patients needing dental emergency treatment in the ongoing pandemic require relevant intra/extraoral dental radiography for adequate diagnosis and treatment planning. METHODS: This article is aimed at adding to the hot debate: Is delay for intraoral radiographs justified or a possible proxy? As a narrative review, it provides an insight into the reasons for delaying intra-oral dental radiographs during in the pandemic and options of the nontraditional radiographic techniques available until the pandemic subsides. Discussion and Conclusion. Cross-contamination concerns through respiratory droplets grow while using intraoral film holders that stimulate gag reflex, coughing, saliva secretion, and if proper disinfection protocols are not applied. Since the patients' acquiring emergency dental treatment cannot be neglected, the return-to-work guidelines by the health regulatory bodies urge to prioritize extraoral radiographic imaging techniques to curb the infection, offering the best diagnostic efficacy. The dental professionals can consider cone-beam computed tomography (CBCT) scans and sectional dental panoramic radiographs (SDPRs), followed by a risk assessment for COVID-19, a safer modality in reducing cross-contamination and assuring an innocuous environment for both patient and coworkers.

Immune checkpoint expression and relationships to anti-PD-L1 immune checkpoint blockade cancer immunotherapy efficacy in aged versus young mice.

Introduction: Aging is the biggest cancer risk, and immune checkpoint (IC) inhibition (ICI) is a revolutionary cancer immunotherapy approach. Nonetheless, there are limited preclinical/clinical data regarding aging effects on ICI outcomes or age effects on IC expression in different organs or tumors. Methods: Flow cytometry assessed IC on immune and non-immune cells in various organs in young and aged BL6 mice. Comparisons: aged versus young naïve WT versus interferon-γ KO mice and WT challenged with B16F10 melanoma and treated with αPD-1 or αPD-L1 ICI. We co-cultured young and aged T cells and myeloid cells in vitro and used OMIQ analyses to test cell-cell interactions. Results: αPD-1 ICI treated melanoma in young and aged hosts, whereas αPD-L1 ICI was only effective in young. We found considerable, previously undescribed age effects on expression of various IC molecules participating in the ICI treatment, including PD-1, PD-L1, PD-L2, and CD80, in distinct organs and in the tumor. These data help explain differential ICI efficacy in young and aged hosts. Host interferon-γ influenced age effects on IC expression in both directions depending on specific IC molecule and tissue. IC expression was further affected by tumor challenge on immune, non-immune, and tumor cells in tumor and other organs. In in vitro co-culture, αPD-1 versus αPD-L1 distinctly influenced polyclonal T cells in young versus aged, suggesting mechanisms for distinct age-related ICI outcomes. Conclusion: Age affects IC expression on specific immune cells in an organ- and tissue-specific manner. ICs were generally higher on aged immune cells. High immune-cell PD-1 could help explain αPD-1 efficacy in aged. High co-expression of CD80 with PD-L1 on dendritic cells could help explain lack of αPD-L1 efficacy in aged hosts. Factors other than myeloid cells and interferon-γ also affect age-related IC expression and T cell function, meriting additional studies.

Rhino-Orbito-Cerebral Mucormycosis During the Second Wave of Covid-19: The Indian Scenario.

Mucormycosis is a life threatening, opportunistic infection often seen in individuals with a weak immune system. With an upsurge of cases of Covid-19, a drastic increase in cases of Rhino-Orbito-Cerebral Mucormycosis is being witnessed at present. This article has been written with the purpose of understanding the factors responsible for it and the challenges it brings along for the Indian health-care system at present. Possible solutions for dealing with these problems have also been included in the manuscript. Google, PubMed and ENT Cochrane databases were searched without a time limit using key words like "Mucormycosis", "Rhino-cerebral-mucormycosis" in conjunction with "COVID-19" and "SARS CoV-2". We found 34 articles to be relevant and hence included them to write this review. Rhino-Orbito-Cerebral Mucormycosis is being seen due to coming together of the three entities-the agent, host and environment that constitute the epidemiological triad for this disease in India. Responsible factors are uncontrolled diabetes mellitus, overzealous use of steroids and antibiotics and other environment related issues. The solutions for these problems lie in spreading awareness about prevention of these practices along with early diagnosis and treatment of mucormycosis. To deal effectively with this situation, particularly when there is an existing overload on otolaryngologists and the rest of the health-care system, a multipronged and multilevel collaborative approach is the need of the hour. With effective Standard Operating Procedures and guidelines promoting a multidisciplinary approach for early diagnosis and treatment, we can surely overcome this situation.

Probiotics maintain the gut microbiome homeostasis during Indian Antarctic expedition by ship.

Ship voyage to Antarctica is a stressful journey for expedition members. The response of human gut microbiota to ship voyage and a feasible approach to maintain gut health, is still unexplored. The present findings describe a 24-day long longitudinal study involving 19 members from 38th Indian Antarctic Expedition, to investigate the impact of ship voyage and effect of probiotic intervention on gut microbiota. Fecal samples collected on day 0 as baseline and at the end of ship voyage (day 24), were analyzed using whole genome shotgun sequencing. Probiotic intervention reduced the sea sickness by 10% compared to 44% in placebo group. The gut microbiome in placebo group members on day 0 and day 24, indicated significant alteration compared to a marginal change in the microbial composition in probiotic group. Functional analysis revealed significant alterations in carbohydrate and amino acid metabolism. Carbohydrate-active enzymes analysis represented functional genes involved in glycoside hydrolases, glycosyltransferases and carbohydrate binding modules, for maintaining gut microbiome homeostasis. Suggesting thereby the possible mechanism of probiotic in stabilizing and restoring gut microflora during stressful ship journey. The present study is first of its kind, providing a feasible approach for protecting gut health during Antarctic expedition involving ship voyage.

Withdrawal Notice: An Insight Data On Medication Safety in Accordance with Drug Complications Based On Automated Drug Dispensing System

The article has been withdrawn at the request of the authors of the journal Current Drug Safety, due to incoherent content.Bentham Science apologizes to the readers of the journal for any inconvenience this may have caused.The Bentham Editorial Policy on Article Withdrawal can be found at https://benthamscience.com/editorial-policies-main.php Bentham Science Disclaimer: It is a condition of publication that manuscripts submitted to this journal have not been published and will not be simultaneously submitted or published elsewhere. Furthermore, any data, illustration, structure or table that has been published elsewhere must be reported, and copyright permission for reproduction must be obtained. Plagiarism is strictly forbidden, and by submit-ting the article for publication the authors agree that the publishers have the legal right to take appropriate action against the authors, if plagiarism or fabricated information is discovered. By submitting a manuscript, the authors agree that the copyright of their article is transferred to the publishers if and when the article is accepted for publication.

Effect of Craniofacial Morphology on Pharyngeal Airway Volume Measured Using Cone-Beam Computed Tomography (CBCT)-A Retrospective Pilot Study.

BACKGROUND: The present study aimed to determine the correlation between pharyngeal airway volume and craniofacial morphology through cone-beam computed tomography (CBCT). Additionally, the study analyzed the influence of gender on pharyngeal airway volume. (2) Methods: 80 CBCT scans of 40 male and 40 female patients (mean age: 15.38 + 1.10 years) fulfilling the eligibility criteria were included. CBCT scans were evaluated for pharyngeal airway volume using the In Vivo Dental 5.1 software. Additionally, CBCT-derived lateral cephalograms were used to assess various craniofacial morphology parameters. To examine the influences of gender on airway volume, T-test was carried out. Correlation between airway volume and craniofacial parameters were measured using Pearson correlation followed by regression analysis. The value of p < 0.05 was considered statistically significant. RESULTS: The mean airway volume was significantly greater in males than in females. A statistically significant negative correlation was found between maxillary plane inclination and pharyngeal airway volume. In contrast, a positive correlation was observed between mandibular length and lower molar inclination with oropharyngeal and total pharyngeal airway volume. Females showed a statistically significant positive correlation between the pharyngeal airway volume and sagittal position of maxilla and mandible; they also showed a negative correlation between oropharyngeal airway volume and the mandibular plane angle. CONCLUSIONS: Overall, the pharyngeal airway space differs significantly between males and females. Craniofacial morphology does have a significant effect on the pharyngeal airway, especially on the oropharyngeal airway volume.

CD122-directed interleukin-2 treatment mechanisms in bladder cancer differ from αPD-L1 and include tissue-selective γδ T cell activation.

BACKGROUND: Anti-programmed death-ligand 1 (αPD-L1) immunotherapy is approved to treat bladder cancer (BC) but is effective in <30% of patients. Interleukin (IL)-2/αIL-2 complexes (IL-2c) that preferentially target IL-2 receptor ß (CD122) augment CD8+ antitumor T cells known to improve αPD-L1 efficacy. We hypothesized that the tumor microenvironment, including local immune cells in primary versus metastatic BC, differentially affects immunotherapy responses and that IL-2c effects could differ from, and thus complement αPD-L1. METHODS: We studied mechanisms of IL-2c and αPD-L1 efficacy using PD-L1+ mouse BC cell lines MB49 and MBT-2 in orthotopic (bladder) and metastatic (lung) sites. RESULTS: IL-2c reduced orthotopic tumor burden and extended survival in MB49 and MBT-2 BC models, similar to αPD-L1. Using antibody-mediated cell depletions and genetically T cell-deficient mice, we unexpectedly found that CD8+ T cells were not necessary for IL-2c efficacy against tumors in bladder, whereas γδ T cells, not reported to contribute to αPD-L1 efficacy, were indispensable for IL-2c efficacy there. αPD-L1 responsiveness in bladder required conventional T cells as expected, but not γδ T cells, altogether defining distinct mechanisms for IL-2c and αPD-L1 efficacy. γδ T cells did not improve IL-2c treatment of subcutaneously challenged BC or orthotopic (peritoneal) ovarian cancer, consistent with tissue-specific and/or tumor-specific γδ T cell contributions to IL-2c efficacy. IL-2c significantly altered bladder intratumoral γδ T cell content, activation status, and specific γδ T cell subsets with antitumor or protumor effector functions. Neither IL-2c nor αPD-L1 alone treated lung metastatic MB49 or MBT-2 BC, but their combination improved survival in both models. Combination treatment efficacy in lungs required CD8+ T cells but not γδ T cells. CONCLUSIONS: Mechanistic insights into differential IL-2c and αPD-L1 treatment and tissue-dependent effects could help develop rational combination treatment strategies to improve treatment efficacy in distinct cancers. These studies also provide insights into γδ T cell contributions to immunotherapy in bladder and engagement of adaptive immunity by IL-2c plus αPD-L1 to treat refractory lung metastases.

Bladder cancer cell-intrinsic PD-L1 signals promote mTOR and autophagy activation that can be inhibited to improve cytotoxic chemotherapy.

Tumor cell-intrinsic programmed death-ligand 1 (PD-L1) signals mediate immunopathologic effects in breast, colon, and ovarian cancers and in melanomas, but bladder cancer (BC) effects are unreported. We show here that BC cell-intrinsic PD-L1 signals in mouse MB49 and human RT4, UM-UC3, and UM-UC-14 BC cells regulate important pathologic pathways and processes, including effects not reported in other cancers. α-PD-L1 antibodies reduced BC cell proliferation in vitro, demonstrating direct signaling effects. BC cell-intrinsic PD-L1 promoted mammalian target of rapamycin complex 1 (mTORC1) signals in vitro and augmented in vivo immune-independent cell growth and metastatic cancer spread, similar to effects we reported in melanoma and ovarian cancer. BC cell-intrinsic PD-L1 signals also promoted basal and stress-induced autophagy, whereas these signals inhibited autophagy in melanoma and ovarian cancer cells. BC cell-intrinsic PD-L1 also mediated chemotherapy resistance to the commonly used BC chemotherapy agents cis-platinum and gemcitabine and to the mTORC1 inhibitor, rapamycin. Thus, BC cell-intrinsic PD-L1 signals regulate important virulence and treatment resistance pathways that suggest novel, actionable treatment targets meriting additional studies. As a proof-of-concept, we showed that the autophagy inhibitor chloroquine improved cis-platinum treatment efficacy in vivo, with greater efficacy in PD-L1 null versus PD-L1-replete BC.

Heat precondition is a potential strategy to combat hepatic injury triggered by severe heat stress.

AIM: Environmental heat stress alters physiological and biochemical functions which leads to multiorgan dysfunction including severe hepatic injury in animals. We hypothesize that heat preconditioning can be potential intervention in combating heat illnesses. MAIN METHODS: Sprague Dawley rats were exposed to moderate heat stress, severe heat stress and heat preconditioning in heat simulation chamber. Mean arterial pressure, heart rate, skin and core temperature were monitored in pre and post heat exposed animals. After stress exposure, blood for hemodynamic and liver tissue for liver function tests, oxidative stress, inflammatory variables and structural studies were collected from rats. Hepatic mitochondria were isolated to study the key structural alterations and functional changes by transmission electron microscopy. KEY FINDINGS: The effect of heat precondition shows improvement in time to attain the core temperature, weight loss, blood pressure and heart rate in rats. Results exhibited decreased levels of liver function tests, elevated levels of free radicals and inflammatory cytokines in heat exposed liver as compared with heat preconditioned animals. Expression levels of mitochondrial heat shock protein 60, superoxide dismutase 1 and uncoupling protein 1 along with activity of electron transport chain complexes I-V were examined and found to be increased in heat preconditioned as compared to heat stressed animals. Morphological studies of liver parenchyma demonstrated reduction in structural deterioration of hepatic lobules and restoration of mitochondrial structural integrity in heat preconditioned rats. SIGNIFICANCE: Present study suggests that heat preconditioning intervention plays a crucial role in protection against heat induced hepatic injury in animals.

Appropriate orthodontic appliances during the COVID-19 pandemic: A scoping review.

INTRODUCTION: The esoteric Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2) infection or COVID-19 has been an unusual plummet in dental/orthodontic practice. Based on current recommendations for various amendments in an orthodontic practice, this scoping review aims to identify orthodontic appliances that are most appropriate to us during this on-going pandemic. METHODS: Electronic databases (PubMed, Scopus, Web of Science, Science Direct, and Google Scholar) were searched up until August 11, 2020. Full-text articles in English with keywords "COVID-19 and Orthodontics" and related search terms were included. RESULTS: Out of 17 retracted articles, only 4 articles were found to be brief the choice for orthodontic appliances in pandemic times speculating clear aligner therapy (CAT) to be a pragmatic solution. The remaining articles were also thoroughly studied and the new norms set by the pandemic were determined. Criteria for orthodontic appliance selection included careful patient screening and collection of records, minimal physical visits, efficient use of technology, virtual consultations but the use of PPE for physical appointments; and lesser AGPs with a lesser risk of airborne transmission. CONCLUSIONS: Subject to regional demands, CAT can be considered as the relatively safer modality-predictable and effective apposite to fixed orthodontic appliances in these unprecedented times.