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Solanum khasianum is a medicinally important plant that is a source of steroidal alkaloids ‘solasodine.’ It has various industrial applications, including oral contraceptives and other pharmaceutical uses. The present study was based on 186 germplasm of S. khasianum, which were analyzed for the stability of economically important traits like solasodine content and fruit yield. The collected germplasm was planted during Kharif 2018, 2019, and 2020 in RCBD with three replications at the experimental farm of CSIR-NEIST, Jorhat, Assam, India. A multivariate approach for stability analysis was applied to identify stable germplasm of S. khasianum for economically important traits. The germplasm was analyzed for additive main effects and multiplicative interaction (AMMI), GGE biplot, multi-trait stability index, and Shukla’s variance which were evaluated for three environments. The AMMI ANOVA revealed significant GE interaction for all the studied traits. The stable and high-yielding germplasm was identified from the AMMI biplot, GGE biplot, Shukla’s variance value, and MTSI plot analysis. Lines no. 90, 85, 70, 107, and 62 were identified as highly stable fruit yielders while, lines no. 1, 146, and 68 were identified as stable high solasodine lines. However, considering both traits, i.e., high fruit yield and solasodine content, MTSI analysis was performed which showed that lines 1, 85, 70,155, 71, 114, 65, 86, 62, 116, 32, and 182 could be used in a breeding program. Thus, this identified germplasm can be considered for further varietal development and could be used in a breeding program. The findings of the present study would be beneficial for the S. khasianum breeding program.

Chronic thromboembolic pulmonary hypertension (CTEPH) is a disease with high mortality and few treatment options. Pulmonary thromboendarterectomy (PTE) is the treatment of choice to relieve pulmonary artery obstruction in patients with chronic thromboembolic pulmonary hypertension (CTEPH). This is a challenging procedure in our perspective because of number of regions such as, late diagnosis, scarcity of appropriate instruments, lack of skilled surgeon as it is not familiar to them. Here we are presenting such a case that was also diagnosed incidentally. We successfully operated the patient with chronic pulmonary thromboembolism with severe pulmonary hypertension. Cardiovasc j 2022; 14(2): 176-179

Small molecule inhibitors of glycosylation enzymes are valuable tools for dissecting glycan functions and potential drug candidates. Screening for inhibitors of glycosyltransferases are mainly performed by in vitro enzyme assays with difficulties moving candidates to cells and animals. Here, we circumvent this by employing a cell-based screening assay using glycoengineered cells expressing tailored reporter glycoproteins. We focused on GalNAc-type O-glycosylation and selected the GalNAc-T11 isoenzyme that selectively glycosylates endocytic low-density lipoprotein receptor (LDLR)-related proteins as targets. Our screen of a limited small molecule compound library did not identify selective inhibitors of GalNAc-T11, however, we identify two compounds that broadly inhibited Golgi-localized glycosylation processes. These compounds mediate the reversible fragmentation of the Golgi system without affecting secretion. We demonstrate how these inhibitors can be used to manipulate glycosylation in cells to induce expression of truncated O-glycans and augment binding of cancer-specific Tn-glycoprotein antibodies and to inhibit expression of heparan sulfate and binding and infection of SARS-CoV-2.

IntroductionAnethole is an industrially important compound which is extensively used in pharmaceuticals, cosmetics, perfumery, food and confectioneries. Anethole is primarily obtained from fennel, anise, and star anise which is highly expensive. Therefore, a study was performed to identify a cost-effective and natural anethole rich strain of Clausena heptaphylla through selection and confirmed through multilocation trial.MethodsThe study was conducted using 23 accessions collected from North eastern region of India from 2014-2018 (initial evaluation trial) and 2018-2022 (multilocation trial). The initial trial was conducted in the experimental farm of CSIR-NEIST, Jorhat, Assam using Complete Randomized Block Design with three replications. Five agronomical traits (plant height, leaf length, leaf width, number of stem branching, herbage yield per plant per cutting) along with essential oil yield and anethole content were evaluated which led to the identification of anethole rich strain of C. heptaphylla. This identified strain was further evaluated along with the two check genotypes for stability based on three yield parameters viz. herbage yield, essential oil yield and anethole content at four multi-locations (Imphal, Jorhat, Runne and Madang) for four years using the same experimental design.Results and discussionThe identified superior line (Jor Lab CH-2) showed consistent performance for the studied yield parameters across all the environments maintaining its superiority. The identified strain exhibited average herbage yield of 1.2 Kg/plant/cutting and essential oil yield of 1.22%. The GC-MS analysis of the essential oil depicted trans anethole as the major constituent (93.25%) followed by estragole (4.85%) while benzene, 1,2-dimethoxy-4-(1-propenyl Isoeugenol methyl ether and cis-anethole were the trace components. This is the first novel report of anethole rich variant of C. heptaphylla which has undergone multilocational trial over the years. Jor Lab CH-2 strain will open a new scope for the industries to isolate anethole from a different source in a cost-effective approach.

The fate of influenza A virus (IAV) infection in the host cell depends on the balance between cellular defence mechanisms and viral evasion strategies. To illuminate the landscape of IAV cellular restriction, we generated and integrated global genetic loss-of-function screens with transcriptomics and proteomics data. Our multi-omics analysis revealed a subset of both IFN-dependent and independent cellular defence mechanisms that inhibit IAV replication. Amongst these, the autophagy regulator TBC1 domain family member 5 (TBC1D5), which binds Rab7 to enable fusion of autophagosomes and lysosomes, was found to control IAV replication in vitro and in vivo and to promote lysosomal targeting of IAV M2 protein. Notably, IAV M2 was observed to abrogate TBC1D5-Rab7 binding through a physical interaction with TBC1D5 via its cytoplasmic tail. Our results provide evidence for the molecular mechanism utilised by IAV M2 protein to escape lysosomal degradation and traffic to the cell membrane, where it supports IAV budding and growth.

Introduction: The human forearm serves an important role in upper extremity function, facilitating placement of the hand in space, thus helping to provide the upper extremity with its unique mobility Fractures involving the bones of forearm present unique problem not encountered with fractures of other long bones. Objective: To evaluate and compare the outcomes of locking compression plate (LCP) with limited contact dynamic compression plate (LC-DCP) for the treatment of adult diaphyseal both bone forearm fractures. Material and Methods: This study conducted in the Department of orthopedics, Mymensingh Medical College Hospital, Mymensingh, Bangladesh from January 2018 to December 2020 included 20 patients with 40 fractures in each group. Patients were selected randomly to receive either LCP (20 patients, 40 fractures) or LC-DCP (20 patients, 40 fractures). Operative time, callus formation, functional outcome and complications were recorded. Results: Mean operative time did not differ significantly in the LCP and LC-DCP group (71.25 and 75.70 minutes respectively) .There was some difference in callus formation and mean time to bone union between the two groups which was significant. However, overall functional outcome did not differ significantly between both the groups. One case had delayed union in the (LC-DCP) group while one patient in each group developed superficial infection. Conclusion: In this prospective study comparing LCP with LC-DCP, the outcomes were equal in terms of final functional outcomes and mean operating time but LCP showed slight advantage in terms of callus formation and mean time to bone union.

Molecular responses to influenza A virus (IAV) infections vary between mammalian species. To identify conserved and species-specific molecular responses, we perform a comparative study of transcriptomic data derived from blood cells, primary epithelial cells, and lung tissues collected from IAV-infected humans, ferrets, and mice. The molecular responses in the human host have unique functions such as antigen processing that are not observed in mice or ferrets. Highly conserved gene coexpression modules across the three species are enriched for IAV infection–induced pathways including cell cycle and interferon (IFN) signaling. TDRD7 is predicted as an IFN-inducible host factor that is up-regulated upon IAV infection in the three species. TDRD7 is required for antiviral IFN response, potentially modulating IFN signaling via the JAK/STAT/IRF9 pathway. Identification of the common and species-specific molecular signatures, networks, and regulators of IAV infection provides insights into host-defense mechanisms and will facilitate the development of novel therapeutic interventions against IAV infection.

Fibrocalculous pancreatic diabetes (FCPD) is a secondary form of diabetes mellitus occurring primarily in tropical countries like Bangladesh and has distinct characteristics. The present study aimed to describe the demographic, clinical and biochemical characteristics of patients with FCPD. This cross-sectional study was conducted at Mymensingh Medical College Hospital, Bangladesh, from January 2019 to December 2021. All patients with FCPD (previously or newly diagnosed) admitted to the inpatient Endocrinology department of the hospital were evaluated. Out of the 15 patients, 73.3% were aged 10-29 years at diagnosis, the male: female ratio was 11:4, rural: urban ratio was 9:6, 20.0% had FCPD in the first-degree family members, 73.3% were underweight, none were overweight/obese or central obese and one of them was hypertensive. Diabetes was uncontrolled in all, with a mean HbA1c of 10.5±1.9%. All but one had low C-peptide and all required insulin to manage diabetes. Although their average (mean or median) lipid parameters were normal, 73.3% of them had dyslipidemia. Among diabetic complications, diabetic nephropathy (66.7%) and neuropathy (66.7%) were more frequent, whereas diabetic retinopathy (6.7%), ischemic heart disease (6.7%) and peripheral vascular disease (6.7%) were less frequently observed; 13.3% had a history of diabetic ketoacidosis. Malnutrition manifested as abnormal skin and hair conditions and anemia was also common in the study subjects. Patients with FCPD are usually young males from rural residences. Microvascular diabetic complications are common, but macrovascular complications and DKA can occur in FCPD.

Electro-deoxidation of ilmenite (FeTiO3) is an economical production method of FeTi, particularly, if the end use is hydrogen storage. In this study, we show that electro-deoxidation of impure FeTiO3 with Ti content lower than Fe, as in the case of low-grade FeTiO3 ore, results in the formation of a two-phase material consisting of FeTi and Fe2Ti. The presence of Fe2Ti is detrimental to the hydrogen storage efficacy. We show for the first time that it is possible to avoid the formation of Fe2Ti or β-Ti as a second phase under similar operating conditions only by tailoring the composition of the cathode precursor, i.e., the addition of TiO2 to low-grade FeTiO3 so that the atomic ratio of Fe:Ti in the precursor is ~ 1:1. Low-grade FeTiO3 with 10 wt% TiO2 resulted in single-phase FeTi with the atomic ratio of Fe:Ti ~ 1:1 in the precursor and in the final reduced alloy. The hydrogen storage capacity of the single-phase FeTi is nearly 36% higher as compared to the two-phase alloy consisting of FeTi–Fe2Ti.

In this work, an electrophoretic deposition technique was used to deposit graphene oxide (GO) on 304 stainless steel. Its corrosion performance was evaluated in a simulated polymer electrolyte membrane fuel cell environment. The corrosion current density (icorr) and interfacial contact resistance (ICR) were measured at 8.9 µA/cm2 and 19.3 mΩ cm2, respectively. The icorr of GO coated 304SS is several orders lower than bare SS 304. Similarly, the ICR of GO coated 304SS is nearly half of bare 304SS at a compaction pressure of 150 N/cm2. The potentiodynamic polarization plot indicates the prevalence of multiple corrosion mechanisms. A prolonged corrosion study for 30 days immersed in the simulated PEM cell environment shows the formation of rounded pits that corroborate the activity of pitting corrosion.

The COVID-19 pandemic is the third outbreak this century of a zoonotic disease caused by a coronavirus, following the emergence of severe acute respiratory syndrome (SARS) in 20031 and Middle East respiratory syndrome (MERS) in 20122. Treatment options for coronaviruses are limited. Here we show that clofazimine-an anti-leprosy drug with a favourable safety profile3-possesses inhibitory activity against several coronaviruses, and can antagonize the replication of SARS-CoV-2 and MERS-CoV in a range of in vitro systems. We found that this molecule, which has been approved by the US Food and Drug Administration, inhibits cell fusion mediated by the viral spike glycoprotein, as well as activity of the viral helicase. Prophylactic or therapeutic administration of clofazimine in a hamster model of SARS-CoV-2 pathogenesis led to reduced viral loads in the lung and viral shedding in faeces, and also alleviated the inflammation associated with viral infection. Combinations of clofazimine and remdesivir exhibited antiviral synergy in vitro and in vivo, and restricted viral shedding from the upper respiratory tract. Clofazimine, which is orally bioavailable and comparatively cheap to manufacture, is an attractive clinical candidate for the treatment of outpatients and-when combined with remdesivir-in therapy for hospitalized patients with COVID-19, particularly in contexts in which costs are an important factor or specialized medical facilities are limited. Our data provide evidence that clofazimine may have a role in the control of the current pandemic of COVID-19 and-possibly more importantly-in dealing with coronavirus diseases that may emerge in the future.

The soaring fidelity of spinel cobaltite system in electrochemistry presents its candidature as an electrode material for high-performance energy storage system and next generation portable devices...

Renal dysfunction following cardiopulmonary bypass is well recognized. The pathophysiology of renal injury is multifactorial and culmination of the interplay of several pathophysiological mechanism. Cardiac surgery requiring cardiopulmonary bypass (CPB) is being carried out on an extended patient population of different age groups and undergoing complex surgical procedure, and thus it places them at an increased risk of renal impairment. Valve replacement surgery is a major and complex surgical procedure requiring CPB. This study was therefore designed to observe the impact of CPB and short term outcome on patients with reduced estimated Glomerular Filtration Rate (eGFR) (60-89ml/min/1.73m²) after valve replacement surgery. This cross-sectional study was conducted in the Department of Cardiac Surgery, National Institute of Cardio Vascular Diseases (NICVD) during the period of January 2015 to August 2016. After fulfillment of enrollment criteria 100 patients were studied for the purpose of the study and they were grouped in two, patients with normal eGFR (≥90ml/min/1.73m²) in Group A and patients with reduced eGFR (60-89ml/min/1.73m²) in Group B. A total of 100 patients, 50 in each group were studied for renal function alteration after valve replacement surgery under cardiopulmonary bypass. The incidence of Acute Kidney Injury (AKI) was higher in Group B. Chi-square test was done and p value was 0.011 which is statistically significant. Postoperative blood loss (ml/hr) (p=0.038), duration of Intensive Care Unit (ICU) stay (in hours) (p=0.009), postoperative hospital stay (days) (p=0.014) was significantly higher and postoperative Urine Output (ml/hr) (p=0.001) was significantly lower in patients with reduced eGFR (60-89ml/min/1.73m²) in Group B. Deterioration of renal function is more in patient with eGFR (60-89ml/min/1.73m²) after valve replacement surgery under cardiopulmonary bypass in comparison with patients with eGFR (≥90ml/min/1.73m²).

The integration of a high atomic percentage (at%) of nitrogen atoms into a graphitic carbon framework is an important issue because of the advantages in improving the functions of nanocarbon materials. Nanoporous graphitic nitrogen-doped carbon hollow spheres are prepared via the direct pyrolysis of nanostructured conducting poly(N-methylaniline) (PNMA) at high temperatures. The synthesized nanocarbon shows up to 8.9 at% doped nitrogen and a high specific surface area of up to 545.1 m2 g−1. The as-prepared nitrogen-doped carbon shows effective oxygen reduction reaction (ORR) activity with a prominent cathodic peak potential of 0.72 V versus a reversible hydrogen electrode (RHE), and good positive onset and half-wave potentials of 0.87 and 0.73 V versus RHE, respectively. The nanoporous graphitic carbon hollow spheres also exhibit a very high specific capacitance of 332.4 F g−1 at a current density of 2 A g−1 and 301.8 F g−1 at a scan rate of 10 mV s−1. Moreover, the prepared nanocarbon displays good rate capabilities and no loss of specific capacitance, even after 5000 cycles of charge–discharge experiments.

Enhanced electrochemical performance of supercapacitors can be achieved through optimal hybridization of electroactive nanomaterials, as it effectively increases the overall surface area and ensures greater electrolyte–electrode interaction. This work reports the realization of a manganese tungstate and amorphous carbon nanotube (MnWO4–aCNT) hybrid and its utilization as the electrodes for a solid-state asymmetric supercapacitor. Large-scale synthesis of aCNTs was carried out via an economical solid-state reaction at low temperature and the walls of these nanotubes were decorated with MnWO4 nanorods via a surfactant-free in situ hydrothermal process. The as-fabricated electrode based on this hybrid over nickel foam delivered a high specific capacitance of 542.18 F g−1 at a scan rate of 2 mV s−1, which is much superior to the values of the structural units separately. This MnWO4-aCNT based electrode showed a high-rate capacity with ∼100% capacitance retention and a coulombic efficiency of ∼100% even after operation for 15 000 cycles. A solid-state asymmetric supercapacitor based on this hybrid attained an energy density of 5.6 W h kg−1 and a power density as high as 893.6 W kg−1. Significantly enhanced electrochemical behaviour registered from the hybrid sample is accounted for by its enhanced surface area and thereby greater number of redox reaction sites along with the positive synergetic effect of the building blocks. This study unlocks further exploration possibilities with other types of aCNT-based hybrid materials for the development of highly stable, non-toxic and cost-effective sustainable energy storage systems.

Background: The angiogenic response to ischemia restores perfusion so as to preserve tissue. A role for mesenchymal-to-endothelial transition in the angiogenic response is controversial. This study is to determine if resident fibroblasts contribute to angiogenesis. Methods: We utilized the murine model of hindlimb ischemia, and in vivo Matrigel plug assay together with lineage tracing studies and single cell RNA-sequencing to examine the transcriptional and functional changes in fibroblasts in response to ischemia. Results: Lineage tracing using Fsp1-Cre: R26R-EYFP mice revealed the emergence within the ischemic hindlimb of a small subset of YFP + CD144 + CD11b − fibroblasts (E* cells) that expressed endothelial cell (EC) genes. Subcutaneous administration of Matrigel in Fsp1-Cre: R26R-EYFP mice generated a plug that became vascularized within 5 days. Isolation of YFP + CD11b - cells from the plug revealed a small subset of YFP + CD144 + CD11b − E* cells which expressed EC genes. Pharmacological or genetic suppression of innate immune signaling reduced vascularity of the Matrigel plug and abrogated the generation of these E* cells. These studies were repeated using human fibroblasts, with fluorescence-activated cell sorting analysis revealing that a small percentage of human fibroblasts that were induced to express EC markers in Matrigel plug assay. Pharmacological suppression or genetic knockout of inflammatory signaling abolished the generation of E* cells, impaired perfusion recovery and increased tissue injury after femoral artery ligation. To further characterize these E* cells, single cell RNA-sequencing studies were performed and revealed 8 discrete clusters of cells expressing characteristic fibroblast genes, of which 2 clusters (C5 and C8) also expressed some EC genes. Ischemia of the hindlimb induced expansion of clusters C5 and C8. The C8 cells did not express CD144, nor did they form networks in Matrigel, but did generate angiogenic cytokines. The C5 fibroblasts most resembled E* cells in their expression of CD144 and their ability to form EC-like networks in Matrigel. Conclusions: Together, these studies indicate the presence of subsets of tissue fibroblasts which seem poised to contribute to the angiogenic response. The expansion of these subsets with ischemia is dependent on activation of innate immune signaling and contributes to recovery of perfusion and preservation of ischemic tissue.

Metallic bipolar plates (BPPs) are potential candidates to replace the conventional graphite BPPs in polymer electrolyte membrane fuel cells (PEMFCs) because they are cost-effective and easy to manufacture. In this present work, we report on the development of Ni–Mo–Cr–P electroplated AISI 1020 steel bipolar plates as a BPP material. The microstructure and composition of the Ni–Mo–Cr–P coatings were optimized varying pH of the electrolyte bath and deposition current. Compared to bare AISI 1020 steel, the coating electroplated at pH 5.5 and a pulse-current density of 0.075 A/cm2 showed an improvement in corrosion resistance by three orders of magnitude. The corrosion current densities of bare and Ni–Mo–Cr–P electroplated AISI 1020 steel in the simulated anodic environment is 1.05 × 10−2 and 4.95 × 10−5 A/cm2, respectively, while that in the simulated cathodic environment it is 3.7 × 10−2 and 4.58 × 10−5 A/cm2, respectively. The enhancement of corrosion resistance is due to the formation of a Mo and Cr based passive oxide layer. The interfacial contact resistance (ICR) of bare and Ni–Mo–Cr–P electroplated AISI 1020 steel is 53.2 and 40.06 mΩ, respectively. ICR of Ni–Mo–Cr–P is lower than the bare AISI 1020 because of the semiconducting nature of the two-layered Mo-based passive oxides. Additionally, Ni–Mo–Cr–P coated AISI 1020 steel shows a higher water contact angle than bare AISI 1020 carbon steel. A single-cell PEMFC stack with Ni–Mo–Cr–P coated AISI 1020 steel BPP shows similar polarization behavior as graphite BPP. However, the performance of Ni–Mo–Cr–P coated AISI 1020 steel BPP lacks durability, and further improvement is necessary before it can be commercially implemented.

Summary The emergence of the novel SARS coronavirus 2 (SARS-CoV-2) in 2019 has triggered an ongoing global pandemic of severe pneumonia-like disease designated as coronavirus disease 2019 (COVID-19)1. The development of a vaccine is likely to require at least 12-18 months, and the typical timeline for approval of a novel antiviral therapeutic can exceed 10 years. Thus, repurposing of known drugs could significantly accelerate the deployment of novel therapies for COVID-19. Towards this end, we profiled a library of known drugs encompassing approximately 12,000 clinical-stage or FDA-approved small molecules. We report the identification of 100 molecules that inhibit viral replication, including 21 known drugs that exhibit dose response relationships. Of these, thirteen were found to harbor effective concentrations likely commensurate with achievable therapeutic doses in patients, including the PIKfyve kinase inhibitor apilimod2–4, and the cysteine protease inhibitors MDL-28170, Z LVG CHN2, VBY-825, and ONO 5334. Notably, MDL-28170, ONO 5334, and apilimod were found to antagonize viral replication in human iPSC-derived pneumocyte-like cells, and the PIKfyve inhibitor also demonstrated antiviral efficacy in a primary human lung explant model. Since most of the molecules identified in this study have already advanced into the clinic, the known pharmacological and human safety profiles of these compounds will enable accelerated preclinical and clinical evaluation of these drugs for the treatment of COVID-19.

Safeguard mechanisms can ameliorate the potential risks associated with cell therapies but currently rely on the introduction of transgenes. This limits their application owing to immunogenicity or transgene silencing. We aimed to create a control mechanism for human cells that is not mediated by a transgene. Using genome editing methods, we disrupt uridine monophosphate synthetase (UMPS) in the pyrimidine de novo synthesis pathway in cell lines, pluripotent cells and primary human T cells. We show that this makes proliferation dependent on external uridine and enables us to control cell growth by modulating the uridine supply, both in vitro and in vivo after transplantation in xenograft models. Additionally, disrupting this pathway creates resistance to 5-fluoroorotic acid, which enables positive selection of UMPS-knockout cells. We envision that this approach will add an additional level of safety to cell therapies and therefore enable the development of approaches with higher risks, especially those that are intended for limited treatment durations. A new safety switch allows for control of cell therapies by dietary uridine.

The production of selective nitrogen-doped graphitic nanoporous carbon plays a pivotal role in performance enhancement and widens the scope of practical applications of functional materials. The sy...

Aphanomyces cochlioides, the causal agent of seedling damping-off and Aphanomyces root rot (ARR) of sugar beet, causes yield losses in major sugar beet growing regions. Currently, a 4-week soil bioassay and a 2-day culture-based assay are used to diagnose presence of A. cochlioides. However, these assays can be time-consuming and lack sensitivity. In this study we developed a sensitive, specific, and rapid assay to detect and quantify DNA of A. cochlioides. We developed a TaqMan qPCR assay targeting a region of the mitochondrial genome of A. cochlioides representing a unique gene order for Aphanomyces with genus-specific primers and a species-specific probe. The qPCR assay detected A. cochlioides in 12 naturally infested field soil samples with disease severity index (DSI) values of 48-100, in sugar beet seedlings 5-7 days after planting, and with as little as 1 fg of pure A. cochlioides DNA. Adult sugar beet roots with ARR symptoms were sampled to further validate this qPCR assay. Aphanomyces cochlioides was detected in 95% of these samples using this qPCR assay, while only 23% of the same samples were positive using a culture-based assay. This shows the improved sensitivity of this qPCR assay for disease diagnosis and could provide growers with ARR risk of a field, which would help them make informed disease management decisions. However, further research is required to translate the results of this study to growers’ fields to quantify A. cochlioides with a high degree of accuracy.

We propose a novel mechanism where Primordial Black Hole (PBH) dark matter is formed much later in the history of the universe between the epoch of Big Bang Nucleosynthesis (BBN) and Cosmic Microwave Background (CMB) photon decoupling. In our setup, one does not need to modify the scale-invariant inflationary power spectra; instead, a late phase transition in strongly interacting fermion-scalar fluid (which naturally occurs around red-shift $ 10^6 \leq \, z_{\scriptscriptstyle T} \, \leq 10^8$ ) creates an instability in the density perturbation as sound speed turns imaginary. As a result, the dark matter perturbation grows exponentially in sub-Compton scales. This follows the immediate formation of early dense dark matter halo, which finally evolves into PBH due to cooling through scalar radiation. We calculate the variance of the density perturbations and PBH fractional abundances $f(M)$ by using a non-monochromatic mass function. We find the peak of our PBH mass function lies between $10^{-16} - 10^{-14}$ solar mass for $ z_{\scriptscriptstyle T} \simeq 10^6$, and thus it can be the entire dark matter of the universe. In PBH formation, one would expect a temporary phase where an attractive scalar balances the Fermi pressure. We numerically confirm that such a state indeed exists, and we find the radius and density profile of the temporary static structure of the dark matter halo, which finally evolves to PBH due to cooling through scalar radiation., 12 pages, 5 figures, Reference modified, Typos Corrected, Accepted for Publication in Astrophysical Journal (ApJ)

NFAT activating protein with ITAM motif 1 (NFAM1) is an ITAM bearing-transmembrane receptor that has been reported to play a role in B cell signaling and development. We performed expression analysis of NFAM1 using publicly available gene expression data sets and found that NFAM1 expression is significantly induced in intestinal biopsies from Crohn’s disease (CD) and ulcerative colitis (UC) patients. At the cellular level, we further observed high expression of NFAM1 in monocytes and neutrophils, and low expression in B and T cells. To explore the role of NFAM1 in multiple immune cells and its potential role in IBD, we generated NFAM1-/- mice. In contrast with previous reports using NFAM1-transgenic mice, NFAM1-/- mice have no obvious defects in immune cell development, or B cell responses. Interestingly, NFAM1-/- monocytes produce reduced levels of TNF-α in response to activation by multiple IBD-relevant stimuli, including CD40L, TLR ligands and MDP. Additional cytokines and chemokines such as IL-6, IL-12, CCL3 and CCL4 are also reduced in CD40L stimulated NFAM1-/- monocytes. Collectively, these findings indicate that NFAM1 promotes monocyte activation, thereby amplifying the response to diverse stimuli. Similarly, we observed that deletion of NFAM1 in human monocytes reduces expression of CD40L-induced CCL4. Lastly, to assess the role of NFAM1 in IBD, we compared development of anti-CD40 induced colitis in NFAM1+/+ and NFAM1-/- mice. We found that although NFAM1 deletion had no impact on development of gut pathology, we did observe a decrease in serum TNF-α, confirming that NFAM1 promotes pro-inflammatory cytokine production in vivo. Taken together, we conclude that NFAM1 functions to amplify cytokine production and should be further evaluated as a therapeutic target for treatment of autoimmune disease.

SUMMARYCyclic GMP-AMP synthase (cGAS) is a primary sensor of aberrant DNA that governs an innate immune signaling cascade, leading to the induction of the type-I interferon response. We have previously identified polyglutamine binding protein 1, PQBP1, as an adaptor molecule required for cGAS-mediated innate immune response of lentiviruses, including the human immunodeficiency virus 1 (HIV-1), but dispensable for the recognition of DNA viruses. HIV-1- encoded DNA is synthesized as a single copy from its RNA genome, and is subsequently integrated into the host chromatin. HIV-1 then produces progeny through amplification and packaging of its RNA genome, thus, in contrast to DNA viruses, HIV-1 DNA is both transient and of low abundance. However, the molecular basis for the detection and verification of this low abundance HIV-1 DNA pathogen-associated molecular pattern (PAMP) is not understood. Here, we elucidate a two-factor authentication strategy that is employed by the innate immune surveillance machinery to selectively respond to the low concentration of PAMP, while discerning these species from extranuclear DNA molecules. We find that, upon HIV-1 infection, PQBP1 decorates intact viral capsid, which serves as a primary verification step for the viral nucleic acid cargo. As the reverse transcription and capsid disassembly initiate, cGAS protein is then recruited to the capsid in a PQBP1-dependent manner, enabling cGAS molecules to be co-positioned at the site of PAMP generation. Thus, these data indicate that PQBP1 recognition of the HIV-1 capsid sanctions a robust cGAS-dependent response to a limited abundance and short-lived DNA PAMP. Critically, this illuminates a molecular strategy wherein the modular recruitment of co-factors to germline encoded pattern recognition receptors (PRRs) serves to enhance repertoire of pathogens that can be sensed by the innate immune surveillance machinery.

Noonan syndrome is a genetic disorder of autosomal dominant inheritance that prevents normal development in various parts of the body. A spontaneous mutation without any family history may also result in the condition. Noonan syndrome can affect normal growth. Birth weight may be normal, but growth slows over time. The growth spurt usually seen during the teenage years may be delayed, and bone maturity also is delayed. In this case A 13 year's male admitted inpatient Department of Endocrinology, Mymensingh Medical College Hospital in April 2021 with not attaining appropriate height and delayed development of secondary sexual characteristics. His birth weight was normal; gestational and neonatal history was uneventful. He was diagnosed with severe pulmonary stenosis at four years and underwent cardiac surgery at his four and eleven years. He was noted to have growth failure from the age of 9 years onward. He had no family history of such type of disease. On examination, he was short statured, underweight, having an upper: lower segment ratio of 1.05 with an arm span of 126cm. He had craniosynostosis, high arched palate, the thick helix of ears (outer rim), small, upturned nose, depressed broad nose, deeply grooved philtrum, keratosis pilaris of the face and upper arm, slant eyes with proptosis, keloid scar over mid-chest, widely spaced nipple, shield chest, pectus excavatum and cubitus valgus. His sexual maturation score was A1, P1, B1. He had pulmonary stenosis with pulmonary hypertension. He had mild microcytic anemia with normal liver, renal, blood glucose, and calcium profile. His bone age was delayed (9 years), thyroid function was normal. The growth hormone dynamic test after clonidine stimulation was normal. His karyotype was 46XY. We have considered giving recombinant growth hormone therapy to accelerate his height.

Immunotherapies have transformed the treatment of cancer through the development of drugs that allow T cells to recognize and kill malignant cells. With this has come a need to evaluate how these drugs affect T cell function. We developed an imaging cytometry-based method that can simultaneously assess T cell activation and the functional ability of these activated T cells to kill cancer cells. We used the Nur77-GFP reporter, which expresses GFP upon T cell receptor signaling, as a readout of T cell activation. We combined this reporter system with an MC38 colon adenocarcinoma cell line expressing a TCR-specific antigen that is used as a model tumor antigen (TA). We show that in T cell-cancer co-culture systems, we are able to quantify the number of activated cytotoxic CD8+ T cells and their ability to recognize and kill cancer cells by imaging cytometry. We tested the sensitivity of this platform by quantifying the proportion of antigen-specific activated T cells in co-cultures with varying ratios of unresponsive T cells, mimicking heterogenous lymphocyte populations. To test our system in a pharmacological context, we evaluated the effects of a SRC family kinase inhibitor on T cell activation and cancer cell death. In each case, we were able to quantify the proportion of activated T cells and the extent of TA-specific cancer cell death. Our findings show that the Nur77-GFP reporter system combined with imaging cytometry is a fast and reproducible system to evaluate how different treatment conditions influence TA recognition by T cells using T cell-cancer co-cultures. The simultaneous analysis of T cell activation with T cell-mediated cancer cell death can be used to rapidly assess immuno-oncology drug candidates and T cell-based therapeutics. Supported by The Elsa U. Pardee Foundation (AHC), the V Foundation Scholar Award (AHC), the Cancer Biology Training Program T32CA009676 (MKC), the Pharmacological Sciences Training Program T32GM140223 (CES), and a Charles W. Edmunds Pharmacology Fellowship (YZ). This study was also supported by resources provided by the U-M Flow Cytometry Core and the U-M Rogel Cancer Center (P30CA046592).

Infection with the novel coronavirus, SARS-CoV-2, results in pneumonia and other respiratory symptoms as well as pathologies at diverse anatomical sites. An outstanding question is whether these diverse pathologies are due to replication of the virus in these anatomical compartments and how and when the virus reaches those sites. To answer these outstanding questions and study the spatiotemporal dynamics of SARS-CoV-2 infection a method for tracking viral spread in vivo is needed. We developed a novel, fluorescently labeled, antibody-based in vivo probe system using the anti-spike monoclonal antibody CR3022 and demonstrated that it could successfully identify sites of SARS-CoV-2 infection in a rhesus macaque model of COVID-19. Our results showed that the fluorescent signal from our antibody-based probe could differentiate whole lungs of macaques infected for 9 days from those infected for 2 or 3 days. Additionally, the probe signal corroborated the frequency and density of infected cells in individual tissue blocks from infected macaques. These results provide proof of concept for the use of in vivo antibody-based probes to study SARS-CoV-2 infection dynamics in rhesus macaques.

Infection with the novel coronavirus, SARS-CoV-2, results in pneumonia and other respiratory symptoms as well as pathologies at diverse anatomical sites. An outstanding question is whether these diverse pathologies are due to replication of the virus in these anatomical compartments and how and when the virus reaches those sites. To answer these outstanding questions and study the spatiotemporal dynamics of SARS-CoV-2 infection a method for tracking viral spread in vivo is needed. We developed a novel, fluorescently labeled, antibody-based in vivo probe system using the anti-spike monoclonal antibody CR3022 and demonstrated that it could successfully identify sites of SARS-CoV-2 infection in a rhesus macaque model of COVID-19. Our results showed that the fluorescent signal from our antibody-based probe could differentiate whole lungs of macaques infected for 9 days from those infected for 2 or 3 days. Additionally, the probe signal corroborated the frequency and density of infected cells in individual tissue blocks from infected macaques. These results provide proof of concept for the use of in vivo antibody-based probes to study SARS-CoV-2 infection dynamics in rhesus macaques.