Mef2c regulates bone mass through Sost-dependent and -independent mechanisms.

Mef2c is a transcription factor that mediates key cellular behaviors that promote endochondral ossification and bone formation. Previously, Mef2c has been shown to regulate Sost transcription via its osteocyte-specific enhancer, ECR5, and conditional deletions of Mef2cfl/fl with either Col1-Cre or Dmp1-Cre produced generalized high bone mass (HBM) consistent with Van Buchem Disease phenotypes. However, Sost-/-; Mef2cfl/fl; Dmp1-Cre mice produced a significantly higher bone mass phenotype that Sost-/- alone suggesting that Mef2c modulates bone mass through additional mechanisms, independent of Sost. To identify new Mef2c transcriptional targets important in bone metabolism, we profiled gene expression by single-cell RNA sequencing in subpopulations of cells isolated from Mef2cfl/fl; Dmp1-Cre and Mef2cfl/fl; Bglap-Cre femurs, both strains exhibiting similar high bone mass phenotypes. However, we found Mef2cfl/fl; Bglap-Cre to also display a growth plate defect characterized by an expansion of several osteoprogenitor subpopulations. Differential gene expression analysis identified a total of 96 up- and 2434 down- regulated genes in Mef2cfl/fl; Bglap-Cre and 176 up- and 1041 down- regulated genes in Mef2cfl/fl; Dmp1-Cre bone cell subpopulations compared to wildtype mice. Mef2c deletion affected the transcriptomes across several cell types including mesenchymal progenitors (MP), osteoprogenitors (OSP), osteoblast (OB), and osteocyte (OCY) subpopulations. Several energy metabolism genes such as Uqcrb, Ndufv2, Ndufs3, Ndufa13, Ndufb9, Ndufb5, Cox6a1, Cox5a, Atp5o, Atp5g2, Atp5b, Atp5 were significantly down regulated in Mef2c-deficient OBs and OCYs, in both strains. Binding motif analysis of promoter regions of differentially expressed genes identified Mef2c binding in Bone Sialoprotein (BSP/Ibsp), a gene known to cause increased trabecular BV/TV in the femurs of Ibsp-/- mice. Immunohistochemical analysis confirmed the absence of Ibsp protein in OBs and OCYs. These findings suggests that the HBM in Sost-/-; Mef2cfl/fl; Dmp1-Cre is caused by a multitude of transcriptional changes in genes that regulate bone formation, two of which are Sost and Ibsp.

Elderly and visually impaired indoor activity monitoring based on Wi-Fi and Deep Hybrid convolutional neural network.

A drop in physical activity and a deterioration in the capacity to undertake daily life activities are both connected with ageing and have negative effects on physical and mental health. An Elderly and Visually Impaired Human Activity Monitoring (EV-HAM) system that keeps tabs on a person's routine and steps in if a change in behaviour or a crisis might greatly help an elderly person or a visually impaired. These individuals may find greater freedom with the help of an EVHAM system. As the backbone of human-centric applications like actively supported living and in-home monitoring for the elderly and visually impaired, an EVHAM system is essential. Big data-driven product design is flourishing in this age of 5G and the IoT. Recent advancements in processing power and software architectures have also contributed to the emergence and development of artificial intelligence (AI). In this context, the digital twin has emerged as a state-of-the-art technology that bridges the gap between the real and virtual worlds by evaluating data from several sensors using artificial intelligence algorithms. Although promising findings have been reported by Wi-Fi-based human activity identification techniques so far, their effectiveness is vulnerable to environmental variations. Using the environment-independent fingerprints generated from the Wi-Fi channel state information (CSI), we introduce Wi-Sense. This human activity identification system employs a Deep Hybrid convolutional neural network (DHCNN). The proposed system begins by collecting the CSI with a regular Wi-Fi Network Interface Controller. Wi-Sense uses the CSI ratio technique to lessen the effect of noise and the phase offset. The t- Distributed Stochastic Neighbor Embedding (t-SNE) is used to eliminate unnecessary data further. The data dimension is decreased, and the negative effects on the environment are eliminated in this process. The resulting spectrogram of the processed data exposes the activity's micro-Doppler fingerprints as a function of both time and location. These spectrograms are put to use in the training of a DHCNN. Based on our findings, EVHAM can accurately identify these actions 99% of the time.

Prevalence of Carotid Artery Stenosis in Patients Undergoing Coronary Artery Bypass Grafting: A Single-Centre Study in Bangladesh.

Coronary artery disease (CAD) is one of the important causes of mortality worldwide. South Asians, notably Indians are unduly prone to develop CAD with its incidence being doubled in the last three decades among both rural and urban settlers. CAD prevalence of in Bangladesh is not known. There are merely a limited number of small-scale epidemiological studies are existing. Recent data indicates CAD prevalence in our country to lie between 1.85-3.4% in rural and 19.6% in an urban sample of working professionals. Despite marked disparity in values, the disease seems to be in rising trend. Patients with concomitant CAD and carotid artery disease are at increased risk of developing peri-operative neurological events including stroke. By far, the prevalence of carotid artery disease in candidates of CABG has not yet been determined in our country. There is a lack of pre-operative guidelines as well for the necessary vascular investigations that should be performed on CABG candidates before they go to the operation table. Pre-operative non-invasive carotid Doppler ultrasonography is a useful screening tool for carotid artery disease in all patients undergoing CABG. This was a cross-sectional observational study, was conducted in the Radiology & Imaging department of Ibrahim Cardiac Hospital & Research Institute, Dhaka, Bangladesh from January 2017 to June 2017. The present cross-sectional study was intended to determine the prevalence of concurrent occurrence of carotid and coronary artery disease in elderly patients undergoing CABG. Total 210 elderly (from 60 & above) patients scheduled for CABG taken as study population. There was bilateral carotid atherosclerotic plaque in 15(12.2%) patients. Right carotid plaque was in 69(56.0%) patients, left carotid plaque in 54(43.9%) patients. Carotid stenosis grading was done in percentage (%). There was significant (>50.0%) stenosis of right carotid system in 12 patients (17.4%) and significant stenosis of left carotid system in 18 patients (33.3%). Right carotid system & bulb was the most common site of plaque formation. We can conclude from this study that a substantial proportion of patients after a particular age possess carotid artery disease simultaneously with coronary artery disease, routine evaluation of carotid arteries of the elderly patients scheduled for CABG is strongly suggested.

IoT-Based Reinforcement Learning Using Probabilistic Model for Determining Extensive Exploration through Computational Intelligence for Next-Generation Techniques.

Computing intelligence is built on several learning and optimization techniques. Incorporating cutting-edge learning techniques to balance the interaction between exploitation and exploration is therefore an inspiring field, especially when it is combined with IoT. The reinforcement learning techniques created in recent years have largely focused on incorporating deep learning technology to improve the generalization skills of the algorithm while ignoring the issue of detecting and taking full advantage of the dilemma. To increase the effectiveness of exploration, a deep reinforcement algorithm based on computational intelligence is proposed in this study, using intelligent sensors and the Bayesian approach. In addition, the technique for computing the posterior distribution of parameters in Bayesian linear regression is expanded to nonlinear models such as artificial neural networks. The Bayesian Bootstrap Deep Q-Network (BBDQN) algorithm is created by combining the bootstrapped DQN with the recommended computing technique. Finally, tests in two scenarios demonstrate that, when faced with severe exploration problems, BBDQN outperforms DQN and bootstrapped DQN in terms of exploration efficiency.

Antibiotic Treatment Prior to Injury Abrogates the Detrimental Effects of LPS in STR/ort Mice Susceptible to Osteoarthritis Development.

Post traumatic osteoarthritis (PTOA) is a form of secondary osteoarthritis (OA) that develops in ~50% of cases of severe articular joint injuries and leads to chronic and progressive degradation of articular cartilage and other joint tissues. PTOA progression can be exacerbated by repeated injury and systemic inflammation. Few studies have examined approaches for blunting or slowing down PTOA progression with emphasis on systemic inflammation; most arthritis studies focused on the immune system have been in the context of rheumatoid arthritis. To examine how the gut microbiome affects systemic inflammation during PTOA development, we used a chronic antibiotic treatment regimen starting at weaning for 6 weeks before anterior cruciate ligament (ACL) rupture in STR/ort mice combined with lipopolysaccharide (LPS)-induced systemic inflammation. STR/ort mice develop spontaneous OA as well as a more severe PTOA phenotype than C57Bl/6J mice. By 6 weeks post injury, histological examination showed a more robust cartilage staining in the antibiotic-treated (AB) STR/ort mice than in the untreated STR/ort controls. Furthermore, we also examined the effects of AB treatment on systemic inflammation and found that the effects of LPS administration before injury are also blunted by AB treatment in STR/ort mice. The AB- or AB+LPS-treated STR/ort injured joints more closely resembled the C57Bl/6J VEH OA phenotypes than the vehicle- or LPS-treated STR/ort, suggesting that antibiotic treatment has the potential to slow disease progression and should be further explored therapeutically as prophylactic post injury. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Degradation-Resistant Hypoxia Inducible Factor-2α in Murine Osteocytes Promotes a High Bone Mass Phenotype.

Molecular oxygen levels vary during development and disease. Adaptations to decreased oxygen bioavailability (hypoxia) are mediated by hypoxia-inducible factor (HIF) transcription factors. HIFs are composed of an oxygen-dependent α subunit (HIF-α), of which there are two transcriptionally active isoforms (HIF-1α and HIF-2α), and a constitutively expressed ß subunit (HIFß). Under normoxic conditions, HIF-α is hydroxylated via prolyl hydroxylase domain (PHD) proteins and targeted for degradation via Von Hippel-Lindau (VHL). Under hypoxic conditions, hydroxylation via PHD is inhibited, allowing for HIF-α stabilization and induction of target transcriptional changes. Our previous studies showed that Vhl deletion in osteocytes (Dmp1-cre; Vhl f/f ) resulted in HIF-α stabilization and generation of a high bone mass (HBM) phenotype. The skeletal impact of HIF-1α accumulation has been well characterized; however, the unique skeletal impacts of HIF-2α remain understudied. Because osteocytes orchestrate skeletal development and homeostasis, we investigated the role of osteocytic HIF-α isoforms in driving HBM phenotypes via osteocyte-specific loss-of-function and gain-of-function HIF-1α and HIF-2α mutations in C57BL/6 female mice. Deletion of Hif1a or Hif2a in osteocytes showed no effect on skeletal microarchitecture. Constitutively stable, degradation-resistant HIF-2α (HIF-2α cDR), but not HIF-1α cDR, generated dramatic increases in bone mass, enhanced osteoclast activity, and expansion of metaphyseal marrow stromal tissue at the expense of hematopoietic tissue. Our studies reveal a novel influence of osteocytic HIF-2α in driving HBM phenotypes that can potentially be harnessed pharmacologically to improve bone mass and reduce fracture risk. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Differential bone adaptation to mechanical unloading and reloading in young, old, and osteocyte deficient mice.

Mechanical unloading causes rapid loss of bone structure and strength, which gradually recovers after resuming normal loading. However, it is not well established how this adaptation to unloading and reloading changes with age. Clinically, elderly patients are more prone to musculoskeletal injury and longer periods of bedrest, therefore it is important to understand how periods of disuse will affect overall skeletal health of aged subjects. Bone also undergoes an age-related decrease in osteocyte density, which may impair mechanoresponsiveness. In this study, we examined bone adaptation during unloading and subsequent reloading in mice. Specifically, we examined the differences in bone adaptation between young mice (3-month-old), old mice (18-month-old), and transgenic mice that exhibit diminished osteocyte density at a young age (3-month-old BCL-2 transgenic mice). Mice underwent 14 days of hindlimb unloading followed by up to 14 days of reloading. We analyzed trabecular and cortical bone structure in the femur, mechanical properties of the femoral cortical diaphysis, osteocyte density and cell death in cortical bone, and serum levels of inflammatory cytokines. We found that young mice lost ~10% cortical bone volume and 27-42% trabecular bone volume during unloading and early reloading, with modest recovery of metaphyseal trabecular bone and near total recovery of epiphyseal trabecular bone, but no recovery of cortical bone after 14 days of reloading. Old mice lost 12-14% cortical bone volume and 35-50% trabecular bone volume during unloading and early reloading but had diminished recovery of trabecular bone during reloading and no recovery of cortical bone. In BCL-2 transgenic mice, no cortical bone loss was observed during unloading or reloading, but 28-31% trabecular bone loss occurred during unloading and early reloading, with little to no recovery during reloading. No significant differences in circulating inflammatory cytokine levels were observed due to unloading and reloading in any of the experimental groups. These results illustrate important differences in bone adaptation in older and osteocyte deficient mice, suggesting a possible period of vulnerability in skeletal health in older subjects during and following a period of disuse that may affect skeletal health in elderly patients.

Observation of Myocardial Involvement in Patients Recovered From COVID-19 by Using Cardiac Magnetic Resonance Imaging, In a Tertiary Care Hospital, Bangladesh.

It was previously reported that coronavirus caused myocardial injury in hospitalized patients. However, delayed cardiac involvement in symptomatic patient recovery from COVID-19 is not yet well known. The objective of this study was to evaluate cardiac involvement by using cardiac magnetic resonance (CMR) in symptomatic post-COVID-19 recovered patients. Thirty (30) patients who recovered from COVID-19 and had recently reported cardiac symptoms were studied in a prospective observational study performed at Popular Medical College Hospital, Dhaka, Bangladesh from March 2021 to September 2021. They underwent CMR examinations. CMR scanning protocol included the following: black blood, cine sequence, both short-axis and long-axis, T2-weight short tau inversion recovery (STIR) sequence, T2- weighted imaging (T2WI) and late gadolinium enhancement (LGE) and quantitative mapping sequences-native T1/T2 mapping and post-contrast T1 mapping. Myocardial edema and late gadolinium enhancement were assessed in all patients. Quantitative evaluation of native T1/T2 and ECV value and cardiac function were evaluated. There were 30 people in all in this study. The average age of the participants in the study was 36.6 years. Fourteen (46.6%) of the patients had abnormal cardiac MRI results, while the remaining 15(53.3%) had negative CMR findings. Among positive findings patients, 8(57.1%) of 14 had increased T2 signal. Increased myocardial edema was found in the same no of patients, involving 53.2% (128 of 224) of LV segments. Only 2 cases (2 of 14) showed mid myocardial and subepicardial LGE, involving 18 of 224, 8.03% of myocardial segments. Global native T1, T2 and ECV values are significantly elevated in all CMR positive findings patients. Native T1 1231ms (IQR: 1281.25-1257.5 versus 1155.5 (IQR: 1137.25-1172.75), T2 40 (IQR: 34.5-43.25) versus 35.5 (IQR: 34-37), ECV 31 (29.75-33.25) versus 23.5 (21.25-24.0), p<0.001; p<0.011 and p<0.001 respectively. Reduced RV functional were found in positive as compared with negative CMR findings patients, EF, 32.05 (IQR: 25.25-39.0) versus 54.5 (IQR: 52.0-57.75) and EDV, 117.5 (IQR: 102.0-134.25) versus 95.0 (IQR: 71.75-99.75), p<0.001 and p<0.001 respectively. In this study cardiac involvement was found in the post-COVID-19 recovered patient with cardiac symptoms. Cardiac MRI findings included myocardial edema, fibrosis and reduced right ventricular function. So attention should be paid to symptomatic post-COVID-19 recovered patients.

Comparison of Microalbuminuria Status in Type 2 Diabetes Mellitus Patients With Chronic Periodontitis: A Cross-Sectional Study.

Background Diabetes has increased the risk for various other ailments in various organs of the body. This can be contributing to periodontitis also as it is the sixth complication related to diabetes mellitus. There is a bidirectional relationship between both. Given the high global prevalence of type-2 diabetes mellitus (T2DM) with periodontitis, it is of great importance to determine the link between periodontitis and microalbuminuria in T2DM patients, which shows early renal disease. Methodology In the present study, a total of 500 patients having T2DM were assessed for periodontitis using Community Periodontal Index (CPI). Anthropometric and biochemical measurements were obtained. Blood samples were estimated for glycemic control tests such as fasting plasma glucose (FPG), glycated hemoglobin (HbA1c), and lipid profile. The subjects who participated in the study were categorized into three groups depending on the albuminuria level. The data were tabulated and analyzed using SPSS Statistics software (IBM Corp., Armonk, USA). Results Out of 500 T2DM subjects, 342 subjects had periodontitis. A statistically significant difference was found in FPG, HbA1c, total cholesterol (TC), triglycerides, and low-density lipoprotein (LDL) between subjects with periodontitis and without periodontitis using a t-test (p = <0.001). The prevalence of normoalbuminuria, micro-, and macroalbuminuria among periodontitis patients was 24.6%, 72.8%, and 2.6% respectively, and the Chi-square analysis revealed that was highly significant. In terms of albuminuria, one-way analysis of variance (ANOVA) revealed statistically significant differences among the periodontitis subjects for the following variables: inputs such as the number of teeth, diabetes mellitus (DM) duration, the level of LDL, and also the depth of the pocket. Intergroup comparison of variables among subjects with albuminuria using the statistical test of Tukey Post Hoc found that there is a significant difference between normoalbuminuria and microalbuminuria. CPI score, tooth mobility, smoking, education level, family income, tooth brushing duration, along with the use of other dental hygiene aids was also found to be statistically significant among subjects with periodontitis. Conclusion The study concluded that T2DM patients had a higher incidence of microalbuminuria among individuals with periodontitis. These subjects also had significantly higher HbA1c and FPG levels than subjects with normoalbuminuria. In addition, subjects with periodontitis exhibited a significant reduction in the total teeth numbers present in the case of albuminuria. The longitudinal correlation between DM, microalbuminuria, and periodontitis could be further investigated in detail to explore possible pathways.

Single-cell RNA-Seq reveals changes in immune landscape in post-traumatic osteoarthritis.

Osteoarthritis (OA) is the most common joint disease, affecting over 300 million people world-wide. Accumulating evidence attests to the important roles of the immune system in OA pathogenesis. Understanding the role of various immune cells in joint degeneration or joint repair after injury is vital for improving therapeutic strategies for treating OA. Post-traumatic osteoarthritis (PTOA) develops in ~50% of individuals who have experienced an articular trauma like an anterior cruciate ligament (ACL) rupture. Here, using the high resolution of single-cell RNA sequencing, we delineated the temporal dynamics of immune cell accumulation in the mouse knee joint after ACL rupture. Our study identified multiple immune cell types in the joint including neutrophils, monocytes, macrophages, B cells, T cells, NK cells and dendritic cells. Monocytes and macrophage populations showed the most dramatic changes after injury. Further characterization of monocytes and macrophages reveled 9 major subtypes with unique transcriptomics signatures, including a tissue resident Lyve1hiFolr2hi macrophage population and Trem2hiFcrls+ recruited macrophages, both showing enrichment for phagocytic genes and growth factors such as Igf1, Pdgfa and Pdgfc. We also identified several genes induced or repressed after ACL injury in a cell type-specific manner. This study provides new insight into PTOA-associated changes in the immune microenvironment and highlights macrophage subtypes that may play a role in joint repair after injury.

Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety.

Background & objectives: Polio, measles, rubella, influenza and rotavirus surveillance programmes are of great public health importance globally. Virus isolation using cell culture is an integral part of such programmes. Possibility of unintended isolation of SARS-CoV-2 from clinical specimens processed in biosafety level-2 (BSL-2) laboratories during the above-mentioned surveillance programmes, cannot be ruled out. The present study was conducted to assess the susceptibility of different cell lines to SARS-CoV-2 used in these programmes. Methods: Replication of SARS-CoV-2 was studied in RD and L20B, Vero/hSLAM, MA-104 and Madin-Darby Canine Kidney (MDCK) cell lines, used for the isolation of polio, measles, rubella, rotavirus and influenza viruses, respectively. SARS-CoV-2 at 0.01 multiplicity of infection was inoculated and the viral growth was assessed by observation of cytopathic effects followed by real-time reverse transcription-polymerase chain reaction (qRT-PCR). Vero CCL-81 cell line was used as a positive control. Results: SARS-CoV-2 replicated in Vero/hSLAM, and MA-104 cells, whereas it did not replicate in L20B, RD and MDCK cells. Vero/hSLAM, and Vero CCL-81 showed rounding, degeneration and detachment of cells; MA-104 cells also showed syncytia formation. In qRT-PCR, Vero/hSLAM and MA-104 showed 106 and Vero CCL-81 showed 107 viral RNA copies per µl. The 50 per cent tissue culture infectious dose titres of Vero/hSLAM, MA-104 and Vero CCL-81 were 105.54, 105.29 and 106.45/ml, respectively. Interpretation & conclusions: Replication of SARS-CoV-2 in Vero/hSLAM and MA-104 underscores the possibility of its unintended isolation during surveillance procedures aiming to isolate measles, rubella and rotavirus. This could result in accidental exposure to high titres of SARS-CoV-2, which can result in laboratory acquired infections and community risk, highlighting the need for revisiting biosafety measures in public health laboratories.

Preexisting Type 1 Diabetes Mellitus Blunts the Development of Posttraumatic Osteoarthritis.

Type 1 diabetes mellitus (T1DM) affects 9.5% of the population. T1DM is characterized by severe insulin deficiency that causes hyperglycemia and leads to several systemic effects. T1DM has been suggested as a risk factor for articular cartilage damage and loss, which could expedite the development of osteoarthritis (OA). OA represents a major public health challenge by affecting 300 million people globally, yet very little is known about the correlation between T1DM and OA. In addition, current studies that have looked at the interaction between diabetes mellitus and OA have reported conflicting results with some suggesting a positive correlation whereas others did not. In this study, we aimed to evaluate whether T1DM exacerbates the development of spontaneous OA or accelerates the progression of posttraumatic osteoarthritis (PTOA) after joint injury. Histological evaluation of T1DM and control joints determined that T1DM mice displayed cartilage degeneration measurements consistent with mild OA phenotypes. RNA sequencing analyses identified significantly upregulated genes in T1DM corresponding to matrix-degrading enzymes known to promote cartilage matrix degradation, suggesting a role of these enzymes in OA development. Next, we assessed whether preexisting T1DM influences PTOA development subsequent to trauma. At 6 weeks post-injury, T1DM injured joints displayed significantly less cartilage damage and joint degeneration than injured non-diabetic joints, suggesting a significant delay in PTOA disease progression. At the single-cell resolution, we identified increased number of cells expressing the chondrocyte markers Col2a1, Acan, and Cytl1 in the T1DM injured group. Our findings demonstrate that T1DM can be a risk factor for OA but not for PTOA. This study provides the first account of single-cell resolution related to T1DM and the risk for OA and PTOA. © 2022 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Influence of various shapes of alumina nanoparticle in integrated polysulfone membrane for separation of lignin from woody biomass and salt rejection.

Lignin valorization is essential in proposing an economic perspective as a raw material for valuable compounds. The bio-refineries require adequate processing to improve the high purity of lignin. Meanwhile, nanofiltration is fascinated attention to obtain high purity value-added products. The effect of alumina nanoparticles on the fabrication of mixed matrix membranes (MMM) has contributed to improvising filtration performance. However, incorporating nanoparticles is a significant issue regarding appropriate size and shape integrated into membrane for better filtration efficiency. The influence of shapes of alumina nanoparticles has been investigated into polysulfone (PSf) membranes for salt and lignin separation. The morphology of alumina was tailored with spindle, cubic, and spherical shapes synthesized at a different calcination temperature of 250, 500, 700 and 900 °C, respectively. The phase transitions were confirmed in X-ray diffraction (XRD) analysis, and the shape of the nanoparticles was observed in a high-resolution transmission electron microscope (HRTEM). The separation efficiency of membranes was tested with salt rejection using sodium sulfate, calcium chloride, potassium sulfate, and sodium chloride. The lignin was extracted from prehydrolysed sawdust, and the synthetic lignosulfonic acid sodium salt solution was separated. The higher lignin rejection of 98.6% and 97.9% were obtained for cubic shaped gamma phase alumina mixed matrix membrane. The high rejection of lignin occurred due to narrow pores channels that could resist the transfer of lignin through the membrane. The results proved that the controllable organization of PSf/alumina mixed matrix membranes could apply for lignocellulose compounds with good efficiency.

Sequential Profiling of Anti-SARS CoV-2 IgG Antibody in Post COVID-19 Patients.

Upon SARS CoV-2 infection, humoral immune system triggers production of anti-SARS CoV-2 IgM and IgG antibodies. Currently, antibodies against SARS CoV-2 spike protein receptor binding domain play a central role in disease protection, making them potential target for in vitro diagnostics applications. This study determines the expression level and sustainability of anti-receptor binding domain (RBD) SARS CoV-2 IgG in post COVID-19 patients. Anti-RBD SARS CoV-2 IgG antibodies in patient serum were analysed by standardised indirect ELISA using SARS CoV-2 spike receptor binding domain protein and HRP conjugated anti-human IgG antibody (anti-h IgG). The study was conducted using 35 adult patient samples with confirmed SARS CoV-2 infection. Additionally, correlation between antibody response after each stage and disease symptoms in post COVID-19 patients were studied. Maximum antibody titre was seen at Day 40 and decreased relatively to Day 180 in antibody positive samples when compared with controls. Overall, more IgG antibody expression is observed in patients who suffered from loss of smell and taste at Day 40. 71% of the positive subjects in this study showed high SARS CoV-2 IgG antibody concentration of above 10 ng/mL and 37% showed strong antibody concentration above 20 ng/mL at the peak of seroconversion.

Immunogenicity of Fractional Dose Inactivated Poliovirus Vaccine in India.

INTRODUCTION: Following the withdrawal of Sabin type 2 from trivalent oral poliovirus vaccine (tOPV) in 2016, the introduction of ≥1 dose of inactivated poliovirus vaccine (IPV) in routine immunization was recommended, either as 1 full dose (0.5mL, intramuscular) or 2 fractional doses of IPV (fIPV-0.1mL, intradermal). India opted for fIPV. We conducted a comparative assessment of IPV and fIPV. METHODS: This was a 4-arm, open-label, multicenter, randomized controlled trial. Infants were enrolled and vaccines administered according to the study design, and the blood was drawn at age 6, 14, and 18 weeks for neutralization testing against all 3 poliovirus types. RESULTS: Study enrolled 799 infants. The seroconversion against type 2 poliovirus with 2 fIPV doses was 85.8% (95% confidence interval [CI]: 80.1%-90.0%) when administered at age 6 and 14 weeks, 77.0% (95% CI: 70.5-82.5) when given at age 10 and 14 weeks, compared to 67.9% (95% CI: 60.4-74.6) following 1 full-dose IPV at age 14 weeks. CONCLUSION: The study demonstrated the superiority of 2 fIPV doses over 1 full-dose IPV in India. Doses of fIPV given at 6 and 14 weeks were more immunogenic than those given at 10 and 14 weeks. Clinical Trial Registry of India (CTRI). Clinical trial registration number was CTRI/2017/02/007793.

Sclerostin Depletion Induces Inflammation in the Bone Marrow of Mice.

Romosozumab, a humanized monoclonal antibody specific for sclerostin (SOST), has been approved for treatment of postmenopausal women with osteoporosis at a high risk for fracture. Previous work in sclerostin global knockout (Sost-/-) mice indicated alterations in immune cell development in the bone marrow (BM), which could be a possible side effect in romosozumab-treated patients. Here, we examined the effects of short-term sclerostin depletion in the BM on hematopoiesis in young mice receiving sclerostin antibody (Scl-Ab) treatment for 6 weeks, and the effects of long-term Sost deficiency on wild-type (WT) long-term hematopoietic stem cells transplanted into older cohorts of Sost-/- mice. Our analyses revealed an increased frequency of granulocytes in the BM of Scl-Ab-treated mice and WT→Sost-/- chimeras, indicating myeloid-biased differentiation in Sost-deficient BM microenvironments. This myeloid bias extended to extramedullary hematopoiesis in the spleen and was correlated with an increase in inflammatory cytokines TNFα, IL-1α, and MCP-1 in Sost-/- BM serum. Additionally, we observed alterations in erythrocyte differentiation in the BM and spleen of Sost-/- mice. Taken together, our current study indicates novel roles for Sost in the regulation of myelopoiesis and control of inflammation in the BM.

Validation of a redesigned pan-poliovirus assay and real-time PCR platforms for the global poliovirus laboratory network.

Surveillance and detection of polioviruses (PV) remain crucial to monitoring eradication progress. Intratypic differentiation (ITD) using the real-time RT-PCR kit is key to the surveillance workflow, where viruses are screened after cell culture isolation before a subset are verified by sequencing. The ITD kit is a series of real-time RT-PCR assays that screens cytopathic effect (CPE)-positive cell cultures using the standard WHO method for virus isolation. Because ITD screening is a critical procedure in the poliovirus identification workflow, validation of performance of real-time PCR platforms is a core requirement for the detection of poliovirus using the ITD kit. In addition, the continual update and improvement of the ITD assays to simplify interpretation in all platforms is necessary to ensure that all real-time machines are capable of detecting positive real-time signals. Four platforms (ABI7500 real-time systems, Bio-Rad CFX96, Stratagene MX3000P, and the Qiagen Rotor-Gene Q) were validated with the ITD kit and a redesigned poliovirus probe. The poliovirus probe in the real-time RT-PCR pan-poliovirus (PanPV) assay was re-designed with a double-quencher (Zen™) to reduce background fluorescence and potential false negatives. The updated PanPV probe was evaluated with a panel consisting of 184 polioviruses and non-polio enteroviruses. To further validate the updated PanPV probe, the new assay was pilot tested in five Global Polio Laboratory Network (GPLN) laboratories (Madagascar, India, Philippines, Pakistan, and Democratic Republic of Congo). The updated PanPV probe performance was shown to reduce background fluorescence and decrease the number of false positives compared to the standard PanPV probe.

Discovery of Small-Molecule Inhibitors of SARS-CoV-2 Proteins Using a Computational and Experimental Pipeline.

A rapid response is necessary to contain emergent biological outbreaks before they can become pandemics. The novel coronavirus (SARS-CoV-2) that causes COVID-19 was first reported in December of 2019 in Wuhan, China and reached most corners of the globe in less than two months. In just over a year since the initial infections, COVID-19 infected almost 100 million people worldwide. Although similar to SARS-CoV and MERS-CoV, SARS-CoV-2 has resisted treatments that are effective against other coronaviruses. Crystal structures of two SARS-CoV-2 proteins, spike protein and main protease, have been reported and can serve as targets for studies in neutralizing this threat. We have employed molecular docking, molecular dynamics simulations, and machine learning to identify from a library of 26 million molecules possible candidate compounds that may attenuate or neutralize the effects of this virus. The viability of selected candidate compounds against SARS-CoV-2 was determined experimentally by biolayer interferometry and FRET-based activity protein assays along with virus-based assays. In the pseudovirus assay, imatinib and lapatinib had IC50 values below 10 µM, while candesartan cilexetil had an IC50 value of approximately 67 µM against Mpro in a FRET-based activity assay. Comparatively, candesartan cilexetil had the highest selectivity index of all compounds tested as its half-maximal cytotoxicity concentration 50 (CC50) value was the only one greater than the limit of the assay (>100 µM).

Single-Cell RNA-Seq Reveals Transcriptomic Heterogeneity and Post-Traumatic Osteoarthritis-Associated Early Molecular Changes in Mouse Articular Chondrocytes.

Articular cartilage is a connective tissue lining the surfaces of synovial joints. When the cartilage severely wears down, it leads to osteoarthritis (OA), a debilitating disease that affects millions of people globally. The articular cartilage is composed of a dense extracellular matrix (ECM) with a sparse distribution of chondrocytes with varying morphology and potentially different functions. Elucidating the molecular and functional profiles of various chondrocyte subtypes and understanding the interplay between these chondrocyte subtypes and other cell types in the joint will greatly expand our understanding of joint biology and OA pathology. Although recent advances in high-throughput OMICS technologies have enabled molecular-level characterization of tissues and organs at an unprecedented resolution, thorough molecular profiling of articular chondrocytes has not yet been undertaken, which may be in part due to the technical difficulties in isolating chondrocytes from dense cartilage ECM. In this study, we profiled articular cartilage from healthy and injured mouse knee joints at a single-cell resolution and identified nine chondrocyte subtypes with distinct molecular profiles and injury-induced early molecular changes in these chondrocytes. We also compared mouse chondrocyte subpopulations to human chondrocytes and evaluated the extent of molecular similarity between mice and humans. This work expands our view of chondrocyte heterogeneity and rapid molecular changes in chondrocyte populations in response to joint trauma and highlights potential mechanisms that trigger cartilage degeneration.

Clear cell odontogenic carcinoma of maxilla: A rare case report.

Clear cell odontogenic carcinoma is a rare, infrequent, aggressive in nature, locally reoccurring odontogenic tumor with a tendency of distant metastasis, occurring during to 4th to 6th decades with a mostly female predilection. Histologically, it is characterized by sheets and islands of vacuolated/clear cells. Till date, only 74 cases have been reported in the literature. We present a case of 45-year-old woman with a tumor mass extending from the maxillary right first premolar till the third molar region measuring 4 cm × 4 cm. The diagnosis was given based on the histopathological findings. Being locally aggressive, the reported data and understanding of this infrequent tumor needs to be strengthened by reporting new cases, and it also demands to be distinguished from other primary and metastatic clear cell tumors of the head-and-neck region.