Generation of a Dcx-CreERT2 knock-in mouse for genetic manipulation of newborn neurons.

A wide variety of CreERT2 driver lines are available for genetic manipulation of adult-born neurons in the mouse brain. These tools have been instrumental in studying fate potential, migration, circuit integration, and morphology of the stem cells supporting lifelong neurogenesis. Despite a wealth of tools, genetic manipulation of adult-born neurons for circuit and behavioral studies has been limited by poor specificity of many driver lines targeting early progenitor cells and by the inaccessibility of lines selective for later stages of neuronal maturation. We sought to address these limitations by creating a new CreERT2 driver line targeted to the endogenous mouse doublecortin locus as a marker of fate-specified neuroblasts and immature neurons. Our new model places a T2A-CreERT2 cassette immediately downstream of the Dcx coding sequence on the X chromosome, allowing expression of both Dcx and CreERT2 proteins in the endogenous spatiotemporal pattern for this gene. We demonstrate that the new mouse line drives expression of a Cre-dependent reporter throughout the brain in neonatal mice and in known neurogenic niches of adult animals. The line has been deposited with the Jackson Laboratory and should provide an accessible tool for studies targeting fate-restricted neuronal precursors.

Selective anti-CXCR2 receptor blockade by AZD5069 inhibits CXCL8-mediated pro-tumorigenic activity in human thyroid cancer cells in vitro.

BACKGROUND: Thyroid cancer is the most common endocrine malignancy. Current therapies are successful, however some patients progress to therapeutically refractive disease. The immunotherapeutic potential of the CXCL8-chemokine/CXCR2-chemokine-receptor system is currently being explored in numerous human cancers. This study aimed to evaluate if the targeting of CXCR2 by its selective antagonist, AZD5069, could modulate CXCL8-mediated pro-tumorigenic effects in thyroid-cancer (TC) cells in vitro. METHODS: Normal human primary thyroid cells (NHT) and TC cell lines TPC-1 (RET/PTC), BCPAP, 8505C and 8305C (BRAFV600e) were treated with AZD5069 (100 pM-10 µM) over a time-course. Viability and proliferation were assessed by WST-1 and crystal violet assays. CXCL8 and CXCR2 mRNA were evaluated by RT-PCR. CXCL8-protein concentrations were measured in cell culture supernatants by ELISA. CXCR2 on cell surface was evaluated by flow-cytometry. Cell-migration was assessed by trans-well-migration chamber-system. RESULTS: AZD5069 exerted negligible effects on cell proliferation or viability. AZD5069 significantly reduced CXCR2, (but not CXCL8) mRNAs in all cell types. CXCR2 was reduced on the membrane of some TC cell lines. A significant reduction of the CXCL8 secretion was found in TPC-1 cells (basal-secretion) and NHT (TNFα-induced secretion). AZD5069 significantly reduced basal and CXCL8-induced migration in NHT and different TC cells. CONCLUSIONS: Our findings confirm the involvement of the CXCL8/CXCR2-axis in promoting pro-tumorigenic effects in TC cells, further demonstrating its immunotherapeutic significance in human cancer.

Scattering of e± by C2H6 Molecule over a Wide Range of Energy: A Theoretical Investigation.

The present work reports the theoretical investigation of the scattering of electrons and positrons by the ethane (C2H6) molecule over the energy range 1 eV-1 MeV. The investigation was carried out by taking into account the screening correction arising from a semiclassical analysis of the atomic geometrical overlapping of the scattering observables calculated in the independent atom approximation. The study is presented through the calculations of a broad spectrum of observable quantities, namely differential, integrated elastic, momentum transfer, viscosity, inelastic, grand total, and total ionization cross-sections and the Sherman functions. A comparative study was carried out between scattering observables for electron impact with those for positron impact to exhibit the similarity and dissimilarity arising out of the difference of the collisions of impinging projectiles with the target. Partial-wave decomposition of the scattering states within the Dirac relativistic framework employing a free-atom complex optical model potential was used to calculate the corresponding observable quantities of the constituent atoms. The results, calculated using our recipe, were compared with the experimental and theoretical works available in the literature. The Sherman function for a e±-C2H6 scattering system is presented for the first time in the literature. The addition of the screening correction to the independent atom approximation method was found to substantially reduce the scattering cross-sections, particularly at forward angles for lower incident energies.

Artificial Intelligence-Controlled Microfluidic Device for Fluid Automation and Bubble Removal of Immunoassay Operated by a Smartphone.

There have been tremendous innovations in microfluidic clinical diagnostics to facilitate novel point-of-care testing (POCT) over the past decades. However, the automatic operation of microfluidic devices that minimize user intervention still lacks reliability and repeatability because microfluidic errors such as bubbles and incomplete filling pose a major bottleneck in commercializing the microfluidic devices for clinical testing. In this work, for the first time, various states of microfluid were recognized to control immunodiagnostics by artificial intelligence (AI) technology. The developed AI-controlled microfluidic platform was operated via an Android smartphone, along with a low-cost polymer device to effectuate enzyme-linked immunosorbent assay (ELISA). To overcome the limited machine-learning capability of smartphones, the region-of-interest (ROI) cascading and conditional activation algorithms were utilized herein. The developed microfluidic chip was incorporated with a bubble trap to remove any bubbles detected by AI, which helps in preventing false signals during immunoassay, as well as controlling the reagents' movement with an on-chip micropump and valve. Subsequently, the developed immunosensing platform was tested for conducting real ELISA using a single microplate from the 96-well to detect the Human Cardiac Troponin I (cTnI) biomarker, with a detection limit as low as 0.98 pg/mL. As a result, the developed platform can be envisaged as an AI-based revolution in microfluidics for point-of-care clinical diagnosis.

HIV-sensitive social protection services in mitigating the challenges and vulnerability of the children affected by HIV/AIDS in Bangladesh: a qualitative study.

Children are particularly vulnerable to the consequences of HIV/AIDS. This paper reports on the challenges faced by children due to their or their parents' HIV/AIDS status and the role of HIV-sensitive social protection in mitigating these challenges. We conducted 32 semi-structured in-depth interviews - 15 with children affected by HIV/AIDS (aged 13-18 years) and 17 with parents of children aged 10-12 years. We found children face significant challenges associated with poverty, gender norms, stigma, and lack of social support which affect their rights to education, shelter, treatment, and food. HIV-sensitive social protection packages such as conditional cash transfer, counselling, and community sensitization help continuation of education, facilitate access to treatment and social protection services, reduce stigma in the community and improve wellbeing of children. Children's accessibility to these services was limited due to distance, transport expenses, poor health conditions of the parents, gender issues, and stigma. HIV-sensitive social protection is effective in improving children's wellbeing. Continuation and expansion of HIV-sensitive social protection packages including conditional cash transfer, psychosocial counselling, and community sensitization and life skills training to facilitate HIV/AIDS affected children's wellbeing and inclusion is recommended.

Exploration of the physical properties of the newly synthesized kagome superconductor LaIr3Ga2 using different exchange-correlation functionals.

LaIr3Ga2 is a kagome superconductor with a superconducting temperature (Tc) of 5.16 K. Here, we present the physical properties of the LaIr3Ga2 kagome superconductor computed via the DFT method wherein six different exchange-correlation functionals were used. The lattice parameters obtained using different functionals are reasonable, with a slight variation compared to experimental values. The bonding nature was explored. The elastic constants (Cij), moduli (B, G, Y), and Vickers hardness (Hv) were computed to disclose the mechanical behavior. The Hv values were estimated to be 2.56-3.16 GPa using various exchange-correlation functionals, indicating the softness of the kagome material. The Pugh ratio, Poisson's ratio, and Cauchy pressure revealed the ductile nature. In addition, mechanical stability was ensured based on the estimated elastic constants. The anisotropic mechanical behavior was confirmed via different anisotropic indices. The Debye temperature (ΘD), melting temperature (Tm), and minimum thermal conductivity (kmin) were calculated to characterize the thermal properties and predict the potential of LaIr3Ga2 as a thermal barrier coating material. The electronic density of states was investigated in detail. The McMillan equation was used to estimate Tc, and the electron-phonon coupling constant (λ) was calculated to explore the superconducting nature. The important optical constants were also calculated to explore its possible optoelectronic applications. The values of reflectivity in the IR-visible region are about 62% to 80%, indicating that the compound under study is suitable as a coating to reduce solar heating. The obtained parameters were compared with previously reported parameters, where available.

Association and expression study of SEPW1 and JAML as preliminary candidate genes related to lamb odor and flavor.

The current study aimed to identify the SEPW1 and JAML genes in lamb as candidate genes related to lamb odor and flavor. The polymorphism study showed that the SEPW1 gene was polymorphic at the BanI restriction site with three genotypes (AA, AG, and GG), whereas the JAML gene was monomorphic at HhaI with genotype (GG). The association of SEPW1 between genotype and lamb odor and flavor (BCFAs and skatole) was analyzed using GLM (General Linear Model). MNA (4-methylnonanoic) was significantly associated (p < 0.05) with lamb odor and flavor. AA genotype has a lower level of MNA than AG and GG, while MOA (4-methyloctanoic), EOA(4-ethyloctanoic), MI (3-methylindole) and MP (3-methylphenol) was not significantly associated with lamb odor and flavor (p > 0.05). Furthermore, to analyze the mRNA expression of SEPW1 in liver tissues, the lambs were divided into three groups based on the genotypes AA, AG, and GG, however, mRNA expression was not differentially expressed between AA, AG, and GG (p > 0.05). These results will enhance the understanding of the functions of SEPW1 gene relation to odor and flavor traits and will shed light on the polymorphism of SEPW1 gene in lamb as a candidate gene for reducing MNA in lamb.

Impact of nitrate and 3-nitrooxypropanol on the carbon footprints of milk from cattle produced in confined-feeding systems across regions in the United States: A life cycle analysis.

It is estimated that enteric methane (CH4) contributes about 70% of all livestock greenhouse gas (GHG) emissions. Several studies indicated that feed additives such as 3-nitrooxypropanol (3-NOP) and nitrate have great potential to reduce enteric emissions. The objective of this study was to determine the net effects of 3-NOP and nitrate on farmgate milk carbon footprint across various regions of the United States and to determine the variability of carbon footprint. A cradle-to-farmgate life cycle assessment was performed to determine regional and national carbon footprint to produce 1 kg of fat- and protein-corrected milk (FPCM). Records from 1,355 farms across 37 states included information on herd structure, milk production and composition, cattle diets, manure management, and farm energy. Enteric CH4, manure CH4, and nitrous oxide were calculated with either the widely used Intergovernmental Panel on Climate Change Tier 2 or region-specific equations available in the literature. Emissions were allocated between milk and meat using a biophysical allocation method. Impacts of nitrate and 3-NOP on baseline regional and national carbon footprint were accounted for using equations adjusted for dry matter intake and neutral detergent fiber. Uncertainty analysis of carbon footprint was performed using Monte Carlo simulations to capture variability due to inputs data. Overall, the milk carbon footprint for the baseline, nitrate, and 3-NOP scenarios were 1.14, 1.09 (4.8% reduction), and 1.01 (12% reduction) kg of CO2-equivalents (CO2-eq)/kg of FPCM across US regions. The greatest carbon footprint for the baseline scenario was in the Southeast (1.26 kg of CO2-eq/kg of FPCM) and lowest for the West region (1.02 kg of CO2-eq/kg of FPCM). Enteric CH4 reductions were 12.4 and 31.0% for the nitrate and 3-NOP scenarios, respectively. The uncertainty analysis showed that carbon footprint values ranged widely (0.88-1.52 and 0.56-1.84 kg of CO2-eq/kg of FPCM within 1 and 2 standard deviations, respectively), suggesting the importance of site-specific estimates of carbon footprint. Considering that 101 billion kilograms of milk was produced by the US dairy industry in 2020, the potential net reductions of GHG from the baseline 117 billion kilograms of CO2-eq were 5.6 and 13.9 billion kilograms of CO2-eq for the nitrate and 3-NOP scenarios, respectively.

Soil organic carbon dynamics in the agricultural soils of Bangladesh following more than 20 years of land use intensification.

Soil organic carbon (SOC) is a key soil quality indicator, as it is a source and storage of plant nutrients and plays a vital role in soil fertility and productivity maintenance. Intensification of agriculture is known to cause SOC decline; however, much of the evidence stems from field-scale experimental trials. The primary aim of this study is to investigate how more than 20 years of agricultural land use intensification in Bangladesh has influenced SOC levels at landscape levels. This was achieved by revisiting in 2012 four sub-sites from the Brahmaputra and Ganges alluviums which were previously sampled (1989-92) by the Soil Resource Development Institute and collecting 190 new samples. These were located at different elevations and subjected to differing amounts of inundation. The SOC was determined using the same method, potassium dichromate wet oxidation, used in the 1989-92 campaign. A comparison of the SOC in the 2012 samples with their historic levels (1989-92) revealed that overall SOC declined significantly across both alluviums as well at their four sub-sites. Further analysis, however, showed that SOC has declined more at higher sites. The higher sites are inundated to a limited level, which makes them suitable for growing multiple crops. Among the land types considered here, the low land sites (because of their topographical position) remain inundated for a greater part of the year, allowing a maximum of only one crop of submerged rice. As a result of reduced biomass decomposition due to anaerobic conditions when inundated, and lower land use/cropping intensity, SOC accretion has occurred in the lower land sites. The SOC levels in South Asian countries are inherently low and agricultural land use intensification fuelled by growing food production demand is causing further SOC loss, which has the potential to jeopardise food security and increase the environmental impact of agriculture.

Positron Emission Intensity in the Decay of 86gY for Use in Dosimetry Studies.

The ß+-emitting radionuclide 86gY (t1/2 = 14.7 h) forms a matched-pair with the ß--emitting therapeutic radionuclide 90Y (t1/2 = 2.7 d) for theranostic application in medicine. This approach demands a precise knowledge of the positron emission probability of the PET nuclide which was until recently rather uncertain for 86gY. In this work, an 86gY source of high radionuclidic purity was prepared and a direct measurement of the positron emission intensity per 100 decay of the parent (hereafter "positron emission intensity") was performed using high-resolution HPGe detector γ-ray spectroscopy. The electron capture intensity was also determined as an additional check by measuring the Kα and Kß X-rays of energies 14.1 and 15.8 keV, respectively, using a low energy HPGe detector. From those measurements, normalized values of 27.2 ± 2.0% for ß+-emission and 72.8 ± 2.0% for EC were obtained. These results are in excellent agreement with values recently reported in the literature based on a detailed decay scheme study.

Phenolic Constituents from Wendlandia tinctoria var. grandis (Roxb.) DC. Stem Deciphering Pharmacological Potentials against Oxidation, Hyperglycemia, and Diarrhea: Phyto-Pharmacological and Computational Approaches.

Wendlandia tinctoria var. grandis (Roxb.) DC. (Family: Rubiaceae) is a semi-evergreen shrub distributed over tropical and subtropical Asia. The present research intended to explore the pharmacological potential of the stem extract of W. tinctoria, focusing on the antioxidant, hypoglycemic, and antidiarrheal properties, and to isolate various secondary metabolites as mediators of such activities. A total of eight phenolic compounds were isolated from the dichloromethane soluble fraction of the stem extract of this plant, which were characterized by electrospray ionization (ESI) mass spectrometric and 1H NMR spectroscopic data as liquiritigenin (1), naringenin (2), apigenin (3), kaempferol (4), glabridin (5), ferulic acid (6), 4-hydroxybenzoic acid (7), and 4-hydroxybenzaldehyde (8). The dichloromethane soluble fraction exhibited the highest phenolic content (289.87 ± 0.47 mg of GAE/g of dried extract) and the highest scavenging activity (IC50 = 18.83 ± 0.07 µg/mL) against the DPPH free radical. All of the isolated compounds, except 4-hydroxybenzaldehyde, exerted a higher antioxidant effect (IC50 = 6.20 ± 0.10 to 16.11 ± 0.02 µg/mL) than the standard butylated hydroxytoluene (BHT) (IC50 = 17.09 ± 0.01 µg/mL). Significant hypoglycemic and antidiarrheal activities of the methanolic crude extract at both doses (200 mg/kg bw and 400 mg/kg bw) were observed in a time-dependent manner. Furthermore, the computational modeling study supported the current in vitro and in vivo findings, and the isolated constituents had a higher or comparable binding affinity for glutathione reductase and urase oxidase enzymes, glucose transporter 3 (GLUT 3), and kappa-opioid receptor, inferring potential antioxidant, hypoglycemic, and antidiarrheal properties, respectively. This is the first report of all of these phenolic compounds being isolated from this plant species and even the first demonstration of the plant stem extract's antioxidant, hypoglycemic, and antidiarrheal potentials. According to the current findings, the W. tinctoria stem could be a potential natural remedy for treating oxidative stress, hyperglycemia, and diarrhea. Nevertheless, further extensive investigation is crucial for thorough phytochemical screening and determining the precise mechanisms of action of the plant-derived bioactive metabolites against broad-spectrum molecular targets.

Smartphone-Based Fully Automated Optofluidic Device with Laser Irradiation-Induced Image Whitening.

Herein, we have developed a fully automated optofluidic device to execute enzyme-linked immunosorbent assay (ELISA) using an active 96-well hybrid lab-on-a-chip (LOC) device. To automate the solution loading into the reaction zone of the device and the post-assayed signal analysis, laser irradiation-induced image whitening was utilized with a smartphone-based optical platform. Two optical phenomena were utilized in our platform to detect the liquid in the reaction chamber using a smartphone. First, by Fresnel's equation, the refraction difference between air and water resulted in the intensity change of the reflected light from the reaction chamber. Therefore, when the liquid was entering into the reaction chamber, the intensity of the reflected light was changed. Second, when the light intensity increases, the smartphone-captured image whitens out due to saturation, even when the red color light was incident. Therefore, by measuring the RGB value of the smartphone image, the intensity changes by the liquid movement in the reaction chamber were successfully monitored. Our platform showed a low detection limit of 7.81 pg/mL for the detection of the NT-proBNP human cardiac biomarker with almost a half standard deviation, compared to the manually operated LOC-based ELISA. As a fully automated LOC adopting a conventional 96-well ELISA platform, we thus concluded that the developed platform can be widely applied for point-of-care clinical tests.

Genetic contributions to autism spectrum disorder.

Autism spectrum disorder (autism) is a heterogeneous group of neurodevelopmental conditions characterized by early childhood-onset impairments in communication and social interaction alongside restricted and repetitive behaviors and interests. This review summarizes recent developments in human genetics research in autism, complemented by epigenetic and transcriptomic findings. The clinical heterogeneity of autism is mirrored by a complex genetic architecture involving several types of common and rare variants, ranging from point mutations to large copy number variants, and either inherited or spontaneous (de novo). More than 100 risk genes have been implicated by rare, often de novo, potentially damaging mutations in highly constrained genes. These account for substantial individual risk but a small proportion of the population risk. In contrast, most of the genetic risk is attributable to common inherited variants acting en masse, each individually with small effects. Studies have identified a handful of robustly associated common variants. Different risk genes converge on the same mechanisms, such as gene regulation and synaptic connectivity. These mechanisms are also implicated by genes that are epigenetically and transcriptionally dysregulated in autism. Major challenges to understanding the biological mechanisms include substantial phenotypic heterogeneity, large locus heterogeneity, variable penetrance, and widespread pleiotropy. Considerable increases in sample sizes are needed to better understand the hundreds or thousands of common and rare genetic variants involved. Future research should integrate common and rare variant research, multi-omics data including genomics, epigenomics, and transcriptomics, and refined phenotype assessment with multidimensional and longitudinal measures.

Electrically or chemically tunable photodetector with ultra high responsivity using graphene/InN nanowire based mixed dimensional barristors.

In this work, an electrically/chemically tunable highly sensitive photodetector based on mixed dimensional heterojunction of graphene and planar InN nanowires (NW) is presented. Controlled partial oxidation of InN has been employed to effectively reduce the high surface carrier concentration of InN, which normally prevents it from forming good rectifying contact with graphene. The resulting surface modified InN NWs have been found to form excellent Schottky junction with graphene, with an increase in effective Schottky barrier height (SBH) by over 1.1 eV and a ratio of forward and reverse bias currents exceeding 4 orders of magnitude. Moreover, very strong barristor (gate tunable heterojunction) action has been observed, withIon/Ioff ≈ 4 orders of magnitude, and SBH increase by >0.3 eV. The barristor has been demonstrated to be highly sensitive to light, especially in the ultra-voilet, visible and near IR spectra. Responsivity was found to be widely tunable by gate voltage, with the highest value exceeding 1000 A W-1. Rise and fall times being in the range of hundreds of ms are indicative of photoconductive gain, which can be attributed to the ultra high responsivity. A method of semi-permanent molecular doping has been demonstrated to realize a two-terminal version of the photodetector, where the desired responsivity can still be achieved without requiring a back gate terminal, enabling the device to be realized on insulating substrates. The effect of encapsulation has been studied as a function of time, which has showed the long term stability of the dopant-induced enhancement and ultra high responsivity of the barristor photodetector.

Assessment of antibacterial, anti-inflammatory, and cytotoxic effects of different extracts of Gynura procumbens leaf.

BACKGROUND: In Southeast Asia, Gynura procumbens has been utilized as a traditional medicinal herb for many diseases. The nontoxic effects of the leaves of G procumbens can be consumed safely for the treatment of many diseases, especially diabetes mellitus. OBJECTIVES: This study aimed to assess the antibacterial, anti-inflammatory, and cytotoxic effects of G procumbens leaves through different extracts. METHODS: Cold extraction was employed for G procumbens leaves. Disc diffusion, bovine serum albumin denaturation, brine shrimp lethality assays, and microscopic examination of tissues (ie, liver, kidney, and heart) were performed to measure antibacterial, anti-inflammatory, and cytotoxic activities, and histopathologic analysis, respectively. RESULTS: The distinct concentrations of aqueous, ethanol, and n-hexane extracts showed prominent antibacterial activity against four pathogenic bacterial strains (Chromobacterium sp, Staphylococcus aureus, Enterococcus faecium, and Escherichia coli 0157:H7:LT). The aqueous extract of G procumbens at a concentration of 200 µg/mL showed potential antibacterial activities against S aureus and E faecium, with mean (SD) zones of inhibition of 15 (1.0) mm and 10 (0.55) mm, respectively. At a concentration of 40 µg/mL, the aqueous extract of G procumbens exhibited significant (P < 0.01) anti-inflammatory activity compared with that of the other 2 extracts in the bovine serum albumin denaturation assay. The n-hexane extract showed moderate cytotoxic activity compared with that of vincristine sulfate. The median lethal concentration values of the aqueous, ethanol, and n-hexane extracts were 427.66, 218.56, and 114.51 µg/mL, respectively. The aqueous extract exhibited lower cytotoxicity than that exhibited by the ethanol and n-hexane extracts. The histopathologic analysis results showed that the aqueous extract of G procumbens had no harmful effects on the different organs of the experimental mice. CONCLUSIONS: The aqueous extract of G procumbens could be a potential source for treating various infectious and chronic diseases. (Curr Ther Res Clin Exp. 2021; 82:XXX-XXX).

Risk factors associated with morbidity and mortality outcomes of COVID-19 patients on the 28th day of the disease course: a retrospective cohort study in Bangladesh.

Diverse risk factors intercede the outcomes of coronavirus disease 2019 (COVID-19). We conducted this retrospective cohort study with a cohort of 1016 COVID-19 patients diagnosed in May 2020 to identify the risk factors associated with morbidity and mortality outcomes. Data were collected by telephone-interview and reviewing records using a questionnaire and checklist. The study identified morbidity and mortality risk factors on the 28th day of the disease course. The majority of the patients were male (64.1%) and belonged to the age group 25-39 years (39.4%). Urban patients were higher in proportion than rural (69.3% vs. 30.7%). Major comorbidities included 35.0% diabetes mellitus (DM), 28.4% hypertension (HTN), 16.6% chronic obstructive pulmonary disease (COPD), and 7.8% coronary heart disease (CHD). The morbidity rate (not-cured) was 6.0%, and the mortality rate (non-survivor) was 2.5%. Morbidity risk factors included elderly (AOR = 2.56, 95% CI = 1.31-4.99), having comorbidity (AOR = 1.43, 95% CI = 0.83-2.47), and smokeless tobacco use (AOR = 2.17, 95% CI = 0.84-5.61). The morbidity risk was higher with COPD (RR = 2.68), chronic kidney disease (CKD) (RR = 3.33) and chronic liver disease (CLD) (RR = 3.99). Mortality risk factors included elderly (AOR = 7.56, 95% CI = 3.19-17.92), having comorbidity (AOR = 5.27, 95% CI = 1.88-14.79) and SLT use (AOR = 1.93, 95% CI = 0.50-7.46). The mortality risk was higher with COPD (RR = 7.30), DM (RR = 2.63), CHD (RR = 4.65), HTN (RR = 3.38), CKD (RR = 9.03), CLD (RR = 10.52) and malignant diseases (RR = 9.73). We must espouse programme interventions considering the morbidity and mortality risk factors to condense the aggressive outcomes of COVID-19.

Calculation of Diamagnetic Susceptibility Tensors of Organic Crystals: From Coronene to Pharmaceutical Polymorphs.

Understanding why crystallization in strong magnetic fields can lead to new polymorphs requires methods to calculate the diamagnetic response of organic molecular crystals. We develop the calculation of the macroscopic diamagnetic susceptibility tensor, χcryst, for organic molecular crystals using periodic density functional methods. The crystal magnetic susceptibility tensor, χcryst, for all experimentally known polymorphs, and its molecular counterpart, χmol, are calculated for flexible pharmaceuticals such as carbamazepine, flufenamic acid, and chalcones, and rigid molecules, such as benzene, pyridine, acridine, anthracene, and coronene, whose molecular magnetic properties have been traditionally studied. A tensor addition method is developed to approximate the crystal diamagnetic susceptibility tensor, χcryst, from the molecular one, χmol, giving good agreement with those calculated directly using the more costly periodic density functional method for χcryst. The response of pharmaceutical molecules and crystals to magnetic fields, as embodied by χcryst, is largely determined by the packing in the crystal, as well as the molecular conformation. The anisotropy of χcryst can vary considerably between polymorphs though the isotropic terms are fairly constant. The implications for developing a computational method for predicting whether crystallization in a magnetic field could produce a novel or different polymorph are discussed.

Therapeutic Effect of Antibiotics Against Escherichia coli O157:H7 in Silk Moth Larvae Animal Model.

The increasing clinical incidence of antibiotic resistance in bacteria is a major global health care issue. Rampant use of antimicrobials is one of the major reasons of the dramatic rise in antibiotic-resistant bacterial strains. Suitable animal models are required to improve our understanding of bacterial pathogenicity, evolution and search for novel antibiotics. The larvae of the silk moth (commonly called silkworm), Bombyx mori, have been used as an animal model for testing the pathogenicity of a clinically isolated strain of enterohemorrhagic Escherichia coli O157:H7 upon injection through hemolymph. Here, we show that a foodborne E. coli O157:H7 strain can kill silkworm larvae upon injection through either hemolymph (blood) or midgut. Bacterial number in the hemolymph started to increase after 3 h of injection into hemolymph, while the number of viable circulating hemocytes decreased. Administration of four well-known antibiotics into the larval hemolymph up to 100 µg per larva showed therapeutic effect with varying efficacies against E. coli O157:H7 with ceftriaxone and imipenem showing better effect. Our findings indicate that silkworm larvae can be used as an animal model to screen for novel antibiotics that are effective against E. coli O157:H7.

Enteric methane, lactation performances, digestibility, and metabolism of nitrogen and energy of Holsteins and Jerseys fed 2 levels of forage fiber from alfalfa silage or corn silage.

Our objective was to determine the effects of replacing alfalfa silage (AS) neutral detergent fiber (NDF) with corn silage (CS) NDF at 2 levels of forage NDF (FNDF) on enteric methane (CH4), lactation performance, ruminal fluid characteristics, digestibility, and metabolism of N and energy in Holstein and Jersey cows. Twelve Holstein and 12 Jersey cows (all primiparous and mid-lactation) were used in a triplicated split-plot 4 × 4 Latin square experiment, where breed and diet formed the main and subplots, respectively. The 4 iso-nitrogenous and iso-starch dietary treatments were arranged as a 2 × 2 factorial with 2 levels of FNDF [19 (low FNDF, LF) and 24% (high FNDF, HF) of dry matter] and 2 sources of FNDF (70:30 and 30:70 ratio of AS NDF to CS NDF). Soyhull (non-forage NDF) and corn grain were respectively used to keep dietary NDF and starch content similar across diets. Total collection of feces and urine over 3 d was performed on 8 cows (1 Latin square from each breed). The difference in dry matter intake (DMI) between Holsteins and Jerseys was greater when fed AS than CS. Compared with Jerseys, Holstein cows had greater body weight (48%), DMI (34%), fat- and protein-corrected milk (FPCM; 31%) and CH4 production (22%; 471 vs. 385 g/d). However, breed did not affect CH4 intensity (g/kg of FPCM) or yield (g/kg of DMI), nutrient digestibility, and N partitioning. Compared with HF, LF-fed cows had greater DMI (10%), N intake (8%), and FPCM (5%), but they were 5% less efficient (both FPCM/DMI and milk N/intake N). Compared with HF, LF-fed cows excreted 11 and 17% less urinary N (g/d and % of N intake, respectively). In spite of lower (2.5%) acetate and higher (10%) propionate (mol/100 mol ruminal volatile fatty acids) LF-fed cows had greater (6%) CH4 production (g/d) than did HF-fed cows, most likely due to increased DMI, as affected mainly by the soyhulls. Compared with AS, CS-fed cows had greater DMI (7%) and FPCM (4%), but they were less efficient (5%), and CH4 yield (g/kg of DMI) was reduced by 8%. In addition, per unit of gross energy intake, CS-fed cows lost less urinary energy (15%) and CH energy (11%) than did AS-fed cows. We concluded that, in contrast to level and source of FNDF, breed did not affect digestive and metabolic efficiencies, and, furthermore, neither breed nor dietary treatments affected CH4 intensity. The tradeoff between CH4 and N losses may have implications in future studies assessing the environmental effects of milk production when approached from a whole-farm perspective.

Short communication: A splice site mutation in CENPU is associated with recessive embryonic lethality in Holstein cattle.

A genome scan for homozygous haplotype deficiency coupled with whole-genome sequence data analysis is a very effective method to identify embryonic lethal mutations in cattle. Among other factors, the power of the approach depends on the availability of a greater amount of genotyping and sequencing data. In the present study, we analyzed the largest known panel of Illumina BovineSNP50 (Illumina Inc., San Diego, CA) genotypes, comprising 401,896 Holstein animals, and we report the mapping of a new embryonic lethal haplotype on chromosome 27, called HH7. We fine mapped the locus in a 2.0-Mb interval using an identical-by-descent approach and analyzed genome sequence data from 4 carrier and 143 noncarrier Holstein bulls to identify the causative mutation. We detected a strong candidate variant in the gene encoding centromere protein U (CENPU), a centromere component essential for proper chromosome segregation during mitosis. The mutant allele is a deletion of 4 nucleotides located at position +3 to +6 bp after the splicing donor site of exon 11. Cross-species nucleotide alignment revealed that the nucleotide at position +3 is entirely conserved among vertebrates, suggesting that it plays an important role in the regulation of CENPU splicing. For verification, we genotyped the candidate variant in 232,775 Holstein individuals and did not observe any homozygotes, whereas 16 were expected (Poisson P-value = 1.1 × 10-7; allele frequency = 0.8%). In addition, genotyping of 250,602 animals from 19 additional breeds revealed that the mutant allele is restricted to animals of Holstein descent. Finally, we estimated the effect of the candidate variant on 2 fertility traits in at-risk mating (i.e., between carrier bulls and daughters of carrier bulls) versus non-risk mating. In agreement with a recessive lethal inheritance pattern, we observed a marked reduction in both conception rate and 56-d nonreturn rate in heifers and cows. The effect on 56-d nonreturn rate suggests that a substantial proportion of homozygous mutants die before 35 d after insemination, which is consistent with the early embryonic death previously reported in CENPU-/- mouse embryos. In conclusion, we demonstrate that with more than 400,000 genotypes, we can map very rare recessive lethal mutations segregating at a frequency below 1% in the population. We recommend performing new analyses regularly as data are accumulating.