Coronavirus: a comparative analysis of detection technologies in the wake of emerging variants.

An outbreak of the coronavirus disease caused by a novel pathogen created havoc and continues to affect the entire world. As the pandemic progressed, the scientific community was faced by the limitations of existing diagnostic methods. In this review, we have compared the existing diagnostic techniques such as reverse transcription polymerase chain reaction (RT-PCR), antigen and antibody detection, computed tomography scan, etc. and techniques in the research phase like microarray, artificial intelligence, and detection using novel materials; on the prospect of sample preparation, detection procedure (qualitative/quantitative), detection time, screening efficiency, cost-effectiveness, and ability to detect different variants. A detailed comparison of different techniques showed that RT-PCR is still the most widely used and accepted coronavirus detection method despite certain limitations (single gene targeting- in context to mutations). New methods with similar efficiency that could overcome the limitations of RT-PCR may increase the speed, simplicity, and affordability of diagnosis. In addition to existing devices, we have also discussed diagnostic devices in the research phase showing high potential for clinical use. Our approach would be of enormous benefit in selecting a diagnostic device under a given scenario, which would ultimately help in controlling the current pandemic caused by the coronavirus, which is still far from over with new variants emerging.

Th1-Th2 and M1-M2 interplay sculpt Aeromonas hydrophila pathogenesis in zebrafish (Danio rerio).

Aeromonas hydrophila is an important aquatic zoonotic pathogen that causes septicemia, necrotizing fasciitis and gastroenteritis in various aquatic and non-aquatic animals. However, the pathogenesis of A. hydrophila is not fully understood. Here, we examined the pathogenicity and histopathology of A. hydrophila in the zebrafish (Danio rerio) model system. We found that the intensity of symptoms and mortality is dose-dependent. Bacterial colonization studies demonstrated that A. hydrophila never cleared out from the fish body but stayed in a state of inactivity till it enters a fresh host. Reinfection studies showed that exposure to A. hydrophila provides immunity against future infection and hence improves fish survival. Gene expression studies revealed the crosstalk between T-helper cell and macrophage responses in fish immune system in response to A. hydrophila and infection memory. Histopathological studies showed that symptoms of tissue damage and inflammation lasted for less duration with less intensity in immunized fish when compared to non-immunized fish. Together, our results suggest that the zebrafish model is a useful system in studying the interplay between A. hydrophila pathogenesis, persistence and immunity.

A Novel Trapezoid Concept of Fibula Cutting Guide for Mandible Angle Reconstruction.

ABSTRACT: Virtual surgical planning (VSP) has revolutionized the planning process in the reconstruction of the mandible with a free fibula flap. The traditional planning and design use a triangular osteotomy guide with a closing wedge movement at the angle. With virtual surgical planning, the authors found an additional lateral mandibular tilt angle and tried to incorporate it in our design of osteotomy guide. The authors have proposed a novel design in a trapezoid shape to accommodate the lateral tilt angle in our osteotomy design. Incorporating the two angles in one angle design improves the aesthesis of the reconstructive procedure and saves operative time.

Assessing the Role of Virtual Surgical Planning in Mandibular Reconstruction With Free Fibula Osteocutaneous Graft.

BACKGROUND: The role of virtual surgical planning and three-dimensional printing in improving the accuracy, precision, functional and aesthetic outcomes have been demonstrated in the literature; however, there is a dearth of studies evaluating these parameters. OBJECTIVE: This study was carried out at a tertiary care center in North India to assess the accuracy of virtual surgical planning in mandibular reconstruction. METHOD: All the included patients were preoperatively assessed with a contrast enhanced computed tomography (CECT) of head and neck. Based on the dimensions a patient-specific fibula cutting guide was fabricated by three-dimensional printing. All patients were evaluated with a postoperative CECT at 6 months of follow-up. The precision outcomes were measured by comparing surgical results with the images of the preoperative virtual planning based on several measurements on the reconstructed mandible such as anteroposterior measurement, transverse measurement, height of the mandible, angle of the mandible, and lateral tilt.The secondary outcomes analyzed were the mean total operative time, mean ischemia time, and morbidity parameters. RESULT: Twelve consecutive patients (6 male, 6 female) with mean age of 33.1 ±â€Š11 years who underwent free fibula graft mandibular reconstruction were included in the study. The mean difference in the anteroposterior, transverse, and height dimensions were 0.87 ±â€Š0.43 mm (P = 0.24), 2.43 ±â€Š3.72 mm (P = 0.64), and 1.08 ±â€Š0.67 (P = 0.88), respectively. The difference in the angles of the reconstructed mandible was 2.37 ±â€Š3.92 degrees (P = 0.51) and of lateral tilt was 1.57 ±â€Š1.5 degrees (P = 0.54). The mean total operative time was 639 ±â€Š27.2 minutes and mean ischemia time was 88.4 ±â€Š8.6 minutes. CONCLUSION: All the patients had satisfactory aesthetic results and good oral function at 6 months postoperative period.

CD44 is required for the pathogenesis of experimental crescentic glomerulonephritis and collapsing focal segmental glomerulosclerosis.

A key feature of glomerular diseases such as crescentic glomerulonephritis and focal segmental glomerulosclerosis is the activation, migration and proliferation of parietal epithelial cells. CD44-positive activated parietal epithelial cells have been identified in proliferative cellular lesions in glomerular disease. However, it remains unknown whether CD44-positive parietal epithelial cells contribute to the pathogenesis of scarring glomerular diseases. Here, we evaluated this in experimental crescentic glomerulonephritis and the transgenic anti-Thy1.1 model for collapsing focal segmental glomerulosclerosis in CD44-deficient (cd44-/-) and wild type mice. For both models albuminuria was significantly lower in cd44-/- compared to wild type mice. The number of glomerular Ki67-positive proliferating cells was significantly reduced in cd44-/- compared to wild type mice, which was associated with a reduced number of glomerular lesions in crescentic glomerulonephritis. In collapsing focal segmental glomerulosclerosis, the extracapillary proliferative cellular lesions were smaller in cd44-/- mice, but the number of glomerular lesions was not different compared to wild type mice. For crescentic glomerulonephritis the influx of granulocytes and macrophages into the glomerulus was similar. In vitro, the growth of CD44-deficient murine parietal epithelial cells was reduced compared to wild type parietal epithelial cells, and human parietal epithelial cell migration could be inhibited using antibodies directed against CD44. Thus, CD44-positive proliferating glomerular cells, most likely parietal epithelial cells, are essential in the pathogenesis of scarring glomerular disease.

Effect of gelatinized-retrograded and extruded starches on characteristics of cookies, muffins and noodles.

The effect of substitution of wheat flour with gelatinized-retrograded starch (GRS) and extruded starch (ES) at 10 and 20 % levels on characteristics of cookies, muffins and noodles was evaluated. Cookies made by substitution of flour with GRS or ES were lighter in color, showed higher spread ratio and resistant starch (RS) content. Muffins made by substitution of flour with GRS or ES were lighter in color, showed less height, specific volume and gas cells and higher RS content. Muffins containing GRS were less firm while those made by incorporating ES showed higher firmness than those made without substitution. Noodles made with substitution of flour with GRS or ES showed higher RS content and reduced water uptake, gruel solid loss, hardness and adhesiveness. Cookies and noodles prepared with and without substitution of flour with GRS or ES did not show any significant differences in terms of overall acceptability scores.

Proximal tubular cells contain a phenotypically distinct, scattered cell population involved in tubular regeneration.

Regeneration of injured tubular cells occurs after acute tubular necrosis primarily from intrinsic renal cells. This may occur from a pre-existing intratubular stem/progenitor cell population or from any surviving proximal tubular cell. In this study, we characterize a CD24-, CD133-, and vimentin-positive subpopulation of cells scattered throughout the proximal tubule in normal human kidney. Compared to adjacent 'normal' proximal tubular cells, these CD24-positive cells contained less cytoplasm, fewer mitochondria, and no brush border. In addition, 49 marker proteins are described that are expressed within the proximal tubules in a similar scattered pattern. For eight of these markers, we confirmed co-localization with CD24. In human biopsies of patients with acute tubular necrosis (ATN), the number of CD24-positive tubular cells was increased. In both normal human kidneys and the ATN biopsies, around 85% of proliferating cells were CD24-positive - indicating that this cell population participates in tubular regeneration. In healthy rat kidneys, the novel cell subpopulation was absent. However, upon unilateral ureteral obstruction (UUO), the novel cell population was detected in significant amounts in the injured kidney. In summary, in human renal biopsies, the CD24-positive cells represent tubular cells with a deviant phenotype, characterized by a distinct morphology and marker expression. After acute tubular injury, these cells become more numerous. In healthy rat kidneys, these cells are not detectable, whereas after UUO, they appeared de novo - arguing against the notion that these cells represent a pre-existing progenitor cell population. Our data indicate rather that these cells represent transiently dedifferentiated tubular cells involved in regeneration.

A Comparison of Antibiotics' Resistance Patterns of E. coli and B. Subtilis in their Biofilms and Planktonic Forms.

BACKGROUND: A biofilm refers to a community of microbial cells that adhere to surfaces that are surrounded by an extracellular polymeric substance. Bacteria employ various defence mechanisms, including biofilm formation, to enhance their survival and resistance against antibiotics. OBJECTIVE: The current study aims to investigate the resistance patterns of Escherichia coli (E. coli) and Bacillus subtilis (B. subtilis) in both biofilms and their planktonic forms. METHODS: E. coli and B. subtilis were used to compare resistance patterns in biofilms versus planktonic forms of bacteria. An antibiotic disc diffusion test was performed to check the resistance pattern of biofilm and planktonic bacteria against different antibiotics such as penicillin G, streptomycin, and ampicillin. Biofilm formation and its validation were done by using quantitative (microtiter plate assay) and qualitative analysis (Congo red agar media). RESULTS: A study of surface-association curves of E. coli and B. subtilis revealed that surface adhesion in biofilms was continuously constant as compared to their planktonic forms, thereby confirming the increased survival of bacteria in biofilms. Also, biofilms have shown high resistance towards the penicillin G, ampicillin and streptomycin as compared to their planktonic form. CONCLUSION: It is safely inferred that E. coli and B. subtilis, in their biofilms, become increasingly resistant to penicillin G, ampicillin and streptomycin.

Mitochondrial electron transport chain in macrophage reprogramming: Potential role in antibacterial immune response.

Macrophages restrain microbial infection and reinstate tissue homeostasis. The mitochondria govern macrophage metabolism and serve as pivot in innate immunity, thus acting as immunometabolic regulon. Metabolic pathways produce electron flows that end up in mitochondrial electron transport chain (mtETC), made of super-complexes regulating multitude of molecular and biochemical processes. Cell-intrinsic and extrinsic factors influence mtETC structure and function, impacting several aspects of macrophage immunity. These factors provide the macrophages with alternate fuel sources and metabolites, critical to gain functional competence and overcoming pathogenic stress. Mitochondrial reactive oxygen species (mtROS) and oxidative phosphorylation (OXPHOS) generated through the mtETC are important innate immune attributes, which help macrophages in mounting antibacterial responses. Recent studies have demonstrated the role of mtETC in governing mitochondrial dynamics and macrophage polarization (M1/M2). M1 macrophages are important for containing bacterial pathogens and M2 macrophages promote tissue repair and wound healing. Thus, mitochondrial bioenergetics and metabolism are intimately coupled with innate immunity. In this review, we have addressed mtETC function as innate rheostats that regulate macrophage reprogramming and innate immune responses. Advancement in this field encourages further exploration and provides potential novel macrophage-based therapeutic targets to control unsolicited inflammation.

Achalasia in Klinefelter syndrome: A suspected pediatric case as well as prevalence analysis suggesting increased risk in this population.

A 4-year-old male with Klinefelter syndrome (KS), speech delay, and intermittent history of coughing and choking during meals was referred for evaluation. Prior evaluation with computed tomography showed a dilated esophagus at the gastroesophageal junction. The patient was unable to tolerate a barium swallow. Upper endoscopy was performed, and an intraoperative esophagogram, demonstrated a "birds beak" appearance suggestive of achalasia. There is no documented relationship between achalasia and KS. However, we utilized TriNetX (a large-scale data clearinghouse) to demonstrate a higher prevalence of achalasia in patients with KS as compared to the general population.

Don't forget to turn around: A case of hematochezia and tenesmus driven by anorectal inflammatory cloacogenic polyp.

We report a case of a 13-year-old male who presented to the Pediatric Gastroenterology clinic with complaints of abdominal pain and frequent stooling, worsened by hematochezia. Despite undergoing endoscopic evaluation twice within a 1-year period, the diagnosis of an Inflammatory Cloacogenic Polyp (ICP) was only revealed during the second evaluation, in which rectal retroflexion was performed. This case highlights the importance of maintaining the ICP at the anorectal transitional zone as part of the differential diagnosis when evaluating patients with symptoms of distal colitis.

NIR-emissive carbon nanodots as a tool to mark ribosomal RNA and nucleolus components using super-resolution microscopy.

Ribosomal RNA (rRNA) plays a key role in protein synthesis and ribosomal biogenesis. The exclusively used commercial dye for RNA staining is SYTO RNASelect, which works in fixed cells only. To overcome this constraint, we synthesized NIR-emissive, highly photostable, and biocompatible carbon nanodots (CNDs) as a fluorescent biomarker for rRNA. The synthesized CNDs could stain rRNA in both live and fixed cells. We were able to visualize rRNA at different sites in eukaryotic cells using super-resolution microscopy (SRM). The CNDs localized rRNA in the dense fibrillar components (DFCs) of the nucleolus, nuclear membrane, and rough endoplasmic reticulum (RER). The super-resolved hollow ring-structured DFC with an FWHM of 140 nm, nuclear membrane with an FWHM of 120 nm, and ER with an FWHM of 115 nm were observed. We further found a marked contrast between the pre-RNA synthesized in cancer cells and normal cells. We believe that these CNDs have great potential in rRNA imaging and comprehending the complex relationships between rRNA dynamics and basic biological processes, disease development, or drug interactions.

A comprehensive review on federated learning based models for healthcare applications.

A disease is an abnormal condition that negatively impacts the functioning of the human body. Pathology determines the causes behind the disease and identifies its development mechanism and functional consequences. Each disease has different identification methods, including X-ray scans for pneumonia, covid-19, and lung cancer, whereas biopsy and CT-scan can identify the presence of skin cancer and Alzheimer's disease, respectively. Early disease detection leads to effective treatment and avoids abiding complications. Deep learning has provided a vast number of applications in medical sectors resulting in accurate and reliable early disease predictions. These models are utilized in the healthcare industry to provide supplementary assistance to doctors in identifying the presence of diseases. Majorly, these models are trained through secondary data sources since healthcare institutions refrain from sharing patients' private data to ensure confidentiality, which limits the effectiveness of deep learning models due to the requirement of extensive datasets for training to achieve optimal results. Federated learning deals with the data in such a way that it doesn't exploit the privacy of a patient's data. In this work, a wide variety of disease detection models trained through federated learning have been rigorously reviewed. This meta-analysis provides an in-depth review of the federated learning architectures, federated learning types, hyperparameters, dataset utilization details, aggregation techniques, performance measures, and augmentation methods applied in the existing models during the development phase. The review also highlights various open challenges associated with the disease detection models trained through federated learning for future research.

Role of UPRmt and mitochondrial dynamics in host immunity: it takes two to tango.

The immune system of a host contains a group of heterogeneous cells with the prime aim of restraining pathogenic infection and maintaining homeostasis. Recent reports have proved that the various subtypes of immune cells exploit distinct metabolic programs for their functioning. Mitochondria are central signaling organelles regulating a range of cellular activities including metabolic reprogramming and immune homeostasis which eventually decree the immunological fate of the host under pathogenic stress. Emerging evidence suggests that following bacterial infection, innate immune cells undergo profound metabolic switching to restrain and countervail the bacterial pathogens, promote inflammation and restore tissue homeostasis. On the other hand, bacterial pathogens affect mitochondrial structure and functions to evade host immunity and influence their intracellular survival. Mitochondria employ several mechanisms to overcome bacterial stress of which mitochondrial UPR (UPRmt) and mitochondrial dynamics are critical. This review discusses the latest advances in our understanding of the immune functions of mitochondria against bacterial infection, particularly the mechanisms of mitochondrial UPRmt and mitochondrial dynamics and their involvement in host immunity.

Integrated point-of-care RT-PCR methods during and after COVID-19 pandemic.

The COVID-19 pandemic has taken the world by surprise and people and organisations worldwide worked in some way or the other to combat the spread; isolate from the infected and get back to normal life, as it was before the pandemic hit. In this regard, the diagnosis of COVID-19 was at the centre of control and prevention and have seen a vehement change in every aspect, especially development of point-of-care testing for better and quick diagnosis. Among different types of techniques developed, the most important was the RT-PCR method of detection which detects nucleic acid of the virus in samples. RT-PCR is a laboratory-based method requiring trained professionals and precise steps for accurate testing. With the advent and spread of the pandemic, number of RT-PCR diagnostic centres rose significantly, and the detection process became less cumbersome, easy to use, ability to handle large volume of samples, more accurate, less time-consuming, and cost-effective. Different industries developed RT-PCR kits, reducing the efforts to prepare laboratory samples. Machines were employed for labour-driven tasks in PCR testing. In addition, new age technologies such as artificial intelligence, IoT, digital systems were combined with RT-PCR for accurate and easy testing. In this review, point-of-care RT-PCR methods, when the COVID-19 started, and the methods now, has been compared on the basis of technological advancements.

ß-Catenin Elicits Drp1-Mediated Mitochondrial Fission Activating the Pro-Apoptotic Caspase-1/IL-1ß Signalosome in Aeromonas hydrophila-Infected Zebrafish Macrophages.

Canonical Wnt signaling plays a major role in regulating microbial pathogenesis. However, to date, its involvement in A. hydrophila infection is not well known. Using zebrafish (Danio rerio) kidney macrophages (ZKM), we report that A. hydrophila infection upregulates wnt2, wnt3a, fzd5, lrp6, and ß-catenin (ctnnb1) expression, coinciding with the decreased expression of gsk3b and axin. Additionally, increased nuclear ß-catenin protein accumulation was observed in infected ZKM, thereby suggesting the activation of canonical Wnt signaling in A. hydrophila infection. Our studies with the ß-catenin specific inhibitor JW67 demonstrated ß-catenin to be pro-apoptotic, which initiates the apoptosis of A. hydrophila-infected ZKM. ß-catenin induces NADPH oxidase (NOX)-mediated ROS production, which orchestrates sustained mitochondrial ROS (mtROS) generation in the infected ZKM. Elevated mtROS favors the dissipation of the mitochondrial membrane potential (ΔΨm) and downstream Drp1-mediated mitochondrial fission, leading to cytochrome c release. We also report that ß-catenin-induced mitochondrial fission is an upstream regulator of the caspase-1/IL-1ß signalosome, which triggers the caspase-3 mediated apoptosis of the ZKM as well as A. hydrophila clearance. This is the first study suggesting a host-centric role of canonical Wnt signaling pathway in A. hydrophila pathogenesis wherein ß-catenin plays a primal role in activating the mitochondrial fission machinery, which actively promotes ZKM apoptosis and helps in containing the bacteria.

Unveiling the Long-Lived Emission of Copper Nanoclusters Embedded in a Protein Scaffold.

Protein-conjugated coinage metal nanoclusters have become promising materials for optoelectronics and biomedical applications. However, the origin of the photoluminescence, especially the long-lived excited state emission in these metal nanoclusters, is still elusive. Here, we unveiled the underlying mechanism of long-lived emission in albumin protein-conjugated copper nanoclusters (Cu NCs) using steady state and time-resolved spectroscopic techniques. Our findings reveal room-temperature phosphorescence (RTP) in protein-conjugated Cu NCs. Time-resolved area-normalized spectra distinguished short- and long-lived components, where the former arises from the singlet state and the latter from the triplet state, thus resulting in RTP. The similarity of the emission spectra at room (298 K) and cryogenic (77 K) temperature ascertains the RTP phenomenon by harvesting the higher-lying triplet states. Time-gated bioimaging of A549 cells using the long-lived emission not only supports RTP emission in the cellular environment but also provides exciting avenues in long-term bioimaging using bovine serum albumin-conjugated Cu NCs.

Tracking the super resolved structure of mitochondria using red emissive carbon nanodots as a fluorescent biomarker.

Herein, we report new red emissive highly photostable and water-soluble carbon nanodots (TPP CNDs) to visualize mitochondrial dynamics using super-resolution radial fluctuations (SRRF) microscopy. The TPP CNDs were synthesized in a one-step method, using 3-(carboxypropyl)triphenylphosphonium bromide (TPP) and o-phenylenediamine (OPDA) as precursors. The obtained crystal structure, NMR, and mass data suggested the presence of [3-(1H-benzimidazol-2-yl)propyl](triphenyl)phosphonium bromide (C28H26N2P+Br-) as a molecular fluorophore (MF) on the surface of the TPP CNDs. The TPP CNDs showed better photostability than the commercially available MitoTracker™ Green and were highly capable for long-term imaging of mitochondrial fission during hyperglycemic conditions and structural changes upon an antidiabetic drug treatment, without altering their fluorescence nature.

Pilot Study of Ondansetron in Improvement of Pediatric Colonoscopy Preparation Outcomes at an Urban Academic Center.

Objectives: To gather initial data on the effectiveness and tolerability of the addition of Ondansetron to bowel preparation regimens to justify a funded, larger, placebo-controlled study. Methods: Design, Setting, and Participants:: A total of 41 pediatric and young adult (age 2-22) patients participated in a single center, open label, parallel randomized trial, with simple randomization. All patients were recruited as outpatients, and all procedures occurred as outpatient procedures, with both recruitment and procedures occurring at a low-resource urban academic medical center in Brooklyn.Interventions and Outcome Measures:: The intervention studied was a single dose of oral-dissolving tablet Ondansetron provided before initiation of bowel preparation using a standardized prep of Polyethylene Glycol 3350 and Bisacodyl. There were 2 arms, a study arm using typical preparation (Polyethylene Glycol 3350 and Bisacodyl) and Ondansetron, and a control arm (Polyethylene Glycol 3350 and Bisacodyl). Patients received standard weight-based dosing. The primary outcome measure assessed was the Boston Bowel Preparation Scale (BBPS) to assess efficacy of preparation. Secondary objectives included evaluation of patient satisfaction via a survey answered by each patient. The questionnaire assessed the presence of the following symptoms during bowel prep: abdominal pain, nausea, bloating, vomiting, scale of ease/difficulty, and if the entire bowel prep was completed. Results: No benefit to BBPS from the addition of Ondansetron to bowel preparation was observed. Statistically significant improvement in reports of abdominal pain (35% decrease in Ondansetron arm) was noted with a P = 0.019. No statistically significant improvement was noted in other symptoms although all domains showed nonsignificant improvement in the Ondansetron arm. Conclusion: No benefit to efficacy of preparation as measured by the BBPS was observed. A single dose of Ondansetron before bowel preparation reduced reports of abdominal pain by 35%, with other symptomatic improvements suggesting possible improvements to be confirmed by a higher-powered study. Trial registration: NCT05439772.

Targeting fuel pocket of cancer cell metabolism: A focus on glutaminolysis.

Advances in cell metabolism over the past few decades have demonstrated glutamine as an essential nutrient for cancer cell survival and proliferation. Glutamine offers a remarkable capacity to fuel diverse metabolic pathways in cancer cells including the Krebs cycle, maintenance of redox homeostasis, and synthesis of cellular building blocks such as nucleic acids, fatty acids, glutathione, and other amino acids. The increase in glutaminolysis has further been linked to the accumulation of oncometabolites such as 2HG (2-Hydroxyglutarate), succinate, fumarate, etc., thereby contributing to tumorigenesis via regulating epigenetic modification of imprinted genes. Therefore, therapeutic targeting of glutaminolysis in cancer cells is worth exploring for possible treatment strategies for cancer management. In this review, we have discussed the detailed mechanism of glutamine uptake, transport, and its instrumental role in rewiring the metabolic adaptation of cancer cells in the tumor microenvironment under nutrient deprivation and hypoxia. Furthermore, we have attempted to provide an updated therapeutic intervention of glutamine metabolism as a treatment strategy for cancer management.