Bacillus subtilis NBRI-W9 simultaneously activates SAR and ISR against Fusarium chlamydosporum NBRI-FOL7 to increase wilt resistance in tomato.

AIMS: The study aimed to determine the pathogenicity of Fusarium species currently prevalent in tomato fields having history of chemical fungicide applications and determine the bio-efficacy of Bacillus subtilis NBRI-W9 as a potent biological control agent. METHODS AND RESULTS: Fusarium was isolated from surface-sterilized infected tomato plants collected from fields. Pathogenicity of 30 Fusarium isolates was determined by in vitro and in vivo assays. Following Koch's postulates, F. chlamydosporum (FOL7) was identified as a virulent pathogen. The biological control of FOL 7 by B. subtilis NBRI-W9 (W9) and the colonization potential of W9 were established using spontaneous rifampicin-resistant mutants. W9 showed 82% inhibition of FOL7 on a dual-culture plate and colonization levels in tomato plants of ∼5.5, ∼3.3, and ∼2.2 log10 CFU/g in root, stem, and leaf tissue, respectively. Antagonistic activity was shown by scanning electron microscopy (SEM) and cell-wall-degradative enzymes. W9 reduced FOL7 infection in net-house and field experiments by 60% and 41%, respectively. Biochemical investigation, defence enzymes, defence gene expression analysis, SEM, and field studies provide evidence of hyperparasitism and induced resistance as the mode of biological control. The study also demonstrates that the potent biocontrol agent W9, isolated from Piper, can colonize tomato plants, control fungal disease by inducing induced systemic resistance (ISR) and systemic acquired resistance (SAR) simultaneously, and increase crop yield by 21.58% under field conditions. CONCLUSIONS: This study concludes that F. chlamydosporum (NBRI-FOL7) is a potent, fungicide-resistant pathogen causing wilt in tomatoes. NBRI-W9 controlled FOL7 through mycoparasitism and simultaneously activated ISR and SAR in plants, providing an attractive tool for disease control that acts at multiple levels.

Trends in Neosubstrate Degradation by Cereblon-Based Molecular Glues and the Development of Novel Multiparameter Optimization Scores.

Molecular glues enable the degradation of previously "undruggable" proteins via the recruitment of cereblon (CRBN) to the target. One major challenge in designing CRBN E3 ligase modulating compounds (CELMoDs) is the selectivity profile toward neosubstrates, proteins recruited by CRBN E3 ligase agents for degradation. Common neosubstrates include Aiolos, Ikaros, GSPT1, CK1α, and SALL4. Unlike achieving potency and selectivity for traditional small molecule inhibitors, reducing the degradation of these neosubstrates is complicated by the ternary nature of the complex formed between the protein, CRBN, and CELMoD. The standard guiding principles of medicinal chemistry, such as enforcing hydrogen bond formation, are less predictive of degradation efficiency and selectivity. Disclosed is an analysis of our glutarimide CELMoD library to identify interpretable chemical features correlated to selectivity profiles and general cytotoxicity. Included is a simple multiparameter optimization function using only three parameters to predict whether molecules will have undesired neosubstrate activity.

Association of acanthosis nigricans with metabolic syndrome - An analytic cross-sectional study

Abstract Background: Globally, few studies have been undertaken to assess the association of acanthosis nigricans (AN) with metabolic syndrome (MS). Most of the available studies have either focused on a particular age group, gender, ethnicity or on a single component of MS. Objectives: To determine the association between AN and MS as a whole and with all individual components of MS in adult patients of either gender. Material and methods: This was a cross-sectional study with a comparative group. Eighty-one subjects were recruited in each group. Fasting plasma glucose (FPG) and lipid profile were done. MS was defined by using the international diabetic federation (IDF) criteria. Association of body mass index (BMI), waist circumference, blood pressure, FPG, high-density lipoprotein (HDL) and triglycerides (TG) with AN was assessed by Pearson's chi-square test followed by univariate and multivariate analysis. Results: The prevalence of MS was found to be significantly higher in the group with AN. On univariate analysis, a significant association of AN was found with BMI, waist circumference, high systolic and diastolic blood pressure, HDL, and TG. Multivariate analysis revealed a significant association between waist circumference, high systolic and diastolic blood pressure, and high TG levels with AN. The risk of MS was found to be eight times higher in cases of AN. Study limitations: The small sample size and single-center data are the limitations of the present study. Conclusion: AN is strongly associated with MS as a whole and with its individual components including increased waist circumference, hypertension, and dyslipidemia.

A scoping review of office-based prostatic stents: past, present, and future of true minimally invasive treatment of benign prostatic hyperplasia.

PURPOSE: To conduct a scoping review of the existing literature and recent developments on prostatic stents for the treatment of benign prostatic hyperplasia (BPH). METHODS: A comprehensive search was performed on Embase, MEDLINE, and Web of Science to identify English literature on prostatic stents for the treatment of BPH. Additional studies and upcoming devices were identified through grey literature search and expert consultation. Study characteristics and stent information were extracted and tabulated narratively. RESULTS: Of the 1171 search results, 64 studies were included in this review. iTiND was the prostatic stent with the most long-term evidence. iTiND is a safe and effective minimally invasive treatment for BPH that preserves sexual function. Adverse events are mild and transitory. Emerging stents (e.g. Zenflow, Butterfly, Urocross, and Exime) had 7/64 eligible studies, where no studies had long-term follow-up. These newer stents show promising results for quality of life and BPH symptom management; however, long-term monitoring and head-to-head comparisons are needed. CONCLUSION: Over the last 50 years, prostatic stents have evolved and demonstrated improved clinical efficacy. iTiND provides a safe and effective outpatient treatment of LUTS secondary to BPH preserving erectile and ejaculatory function. Emerging prostatic stents are a promising, effective, and safe intervention in well-selected patients interested in its benefits.

Association of acanthosis nigricans with metabolic syndrome - An analytic cross-sectional study.

BACKGROUND: Globally, few studies have been undertaken to assess the association of acanthosis nigricans (AN) with metabolic syndrome (MS). Most of the available studies have either focused on a particular age group, gender, ethnicity or on a single component of MS. OBJECTIVES: To determine the association between AN and MS as a whole and with all individual components of MS in adult patients of either gender. MATERIAL AND METHODS: This was a cross-sectional study with a comparative group. Eighty-one subjects were recruited in each group. Fasting plasma glucose (FPG) and lipid profile were done. MS was defined by using the international diabetic federation (IDF) criteria. Association of body mass index (BMI), waist circumference, blood pressure, FPG, high-density lipoprotein (HDL) and triglycerides (TG) with AN was assessed by Pearson's chi-square test followed by univariate and multivariate analysis. RESULTS: The prevalence of MS was found to be significantly higher in the group with AN. On univariate analysis, a significant association of AN was found with BMI, waist circumference, high systolic and diastolic blood pressure, HDL, and TG. Multivariate analysis revealed a significant association between waist circumference, high systolic and diastolic blood pressure, and high TG levels with AN. The risk of MS was found to be eight times higher in cases of AN. STUDY LIMITATIONS: The small sample size and single-center data are the limitations of the present study. CONCLUSION: AN is strongly associated with MS as a whole and with its individual components including increased waist circumference, hypertension, and dyslipidemia.

A Systematic Review on Participant Diversity in Clinical Trials-Have We Made Progress for the Management of Obesity and Its Metabolic Sequelae in Diet, Drug, and Surgical Trials.

OBJECTIVE: Individuals from Black and Hispanic backgrounds represent a minority of the overall US population, yet are the populations most affected by the disease of obesity and its comorbid conditions. Black and Hispanic individuals remain underrepresented among participants in obesity clinical trials, despite the mandate by the National Institutes of Health (NIH) Revitalization Act of 1993. This systematic review evaluates the racial, ethnic, and gender diversity of clinical trials focused on obesity at a national level. METHODS: Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a systematic review of clinicaltrials.gov, PubMed, Cochrane Central, and Web of Science was undertaken to locate phase 3 and phase 4 clinical trials on the topic of obesity that met associated inclusion/exclusion criteria. Ultimately, 18 studies were included for review. RESULTS: White non-Hispanic individuals represented the majority of clinical trial participants, as did females. No study classified participants by gender identity. Reporting of race/ethnicity was not uniform, with noted variability among racial/ethnic subgroups. CONCLUSIONS: Our findings suggest that disparities remain in the diverse racial, ethnic, and gender representation of participants engaged in clinical trials on obesity relative to the prevalence of obesity in underrepresented populations. Commitment to inclusive and intentional recruiting practices is needed to increase the representation of underrepresented groups, thus increasing the generalizability of future research.

Synergistic action of Trichoderma koningiopsis and T. asperellum mitigates salt stress in paddy.

Intensive cultivation increases the salinity and alkalinity of soil leading to its degradation. Such soil lead to abiotic stress conditions in plants causing ROS-mediated cellular damage. Microbes constitute an important group of bio-stimulants, which are promising alternatives to reduce ROS-mediated abiotic stresses and improve plant growth. In the present study synergistic activity of stress-tolerant Trichoderma koningiopsis NBRI-PR5 (MTCC 25372) and T. asperellum NBRI-K14 (MTCC 25373) (TrichoMix) was assessed in paddy crop under salt stress conditions. Improved soil microbial biomass carbon (MBC), total organic carbon (TOC), and available nutrients N/P/K by 2-3 folds was observed in the pot experiment using the TrichoMix. It restored the heterogeneous microbial population of the paddy rhizosphere during salt stress and modulated the soil enzyme activities. The anatomical distortions in rice roots due to salt stress were stabilized in presence of the TrichoMix. Different stress marker genes (OsMAPK5, OsAPX, OsGST, OsUSP, OsBADH, OsLYSO, OsNRAMP6, and OsBz8) were differentially modulated by the TrichoMix in presence of salt stress as compared to the control. The TrichoMix increased the yield by 10% in marginally stressed fields; however, it enhanced the yield by approximately 60% when used with the 50% recommended dose of NPK. In the integrated treatment, Fe and Zn were fortified by approximately 40% and 29% respectively in the grains. From the present study, it was concluded that the TrichoMix stimulated the rice plants to accumulate osmoprotectants, improved the anatomical features, modulated the plant defense system, and improved the grain yield and quality. Therefore, the NBRI-PR5 and NBRI-K14 mixture may be used as a bio-stimulant to increase productivity in the rapidly deteriorating soil and reduce the NPK inputs. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01192-6.

Understanding the Morphological Evolution of InSb Nanoflags Synthesized in Regular Arrays by Chemical Beam Epitaxy.

InSb nanoflags are grown by chemical beam epitaxy in regular arrays on top of Au-catalyzed InP nanowires synthesized on patterned SiO2/InP(111)B substrates. Two-dimensional geometry of the nanoflags is achieved by stopping the substrate rotation in the step of the InSb growth. Evolution of the nanoflag length, thickness and width with the growth time is studied for different pitches (distances in one of the two directions of the substrate plane). A model is presented which explains the observed non-linear time dependence of the nanoflag length, saturation of their thickness and gradual increase in the width by the shadowing effect for re-emitted Sb flux. These results might be useful for morphological control of InSb and other III-V nanoflags grown in regular arrays.

Impact of Gd-153 scanning line source attenuation correction on downstream invasive testing.

BACKGROUND: Attenuation correction (AC) using hardware and software solutions has been shown to increase the specificity of SPECT MPI by decreasing false positive results and improving prognostic ability. Theoretically this should reduce downstream testing and unnecessary costs. We sought to assess the consequences of the use of Gd-153 scanning line source attenuation correction during SPECT myocardial perfusion imaging (MPI) on downstream invasive testing. METHODS: All patients who underwent a clinically indicated Tc-99m stress SPECT MPI study from 2013 to 2015 at five hospitals (2 with AC and 3 without) were retrospectively reviewed. Patient demographics, results of testing, subsequent coronary angiography within 3 months, and revascularization were recorded. The results of the MPI studies, downstream angiogram utilization, and results of angiography were compared and a propensity matched subgroup analysis was performed. RESULTS: A total of 9968 patients underwent SPECT MPI during the study time period (6106 performed with AC and 3862 without). Out of 3928 patients included in the propensity matched cohort, there was no difference in the proportion of abnormal MPI results between the two groups (31.5% vs 30.4%, P = 0.47), however, more patients underwent coronary angiography within 90 days in the AC group (10.6% vs 8.7%, P = 0.05). There was no significant difference in the proportion of patients with angiographically significant obstructive disease (53.4% vs 56.1%, P = 0.19), however, fewer patients in the AC group with obstructive coronary disease were revascularized (36.1% vs 46.8%, P = 0.04). The findings remained consistent after sub-group analysis in patients without known coronary disease. CONCLUSION: The use of scanning line source AC did not meaningfully influence the rate of abnormal MPI results or downstream invasive testing in this cohort. The clinical utility of scanning line source AC may be limited to facilitating stress-first imaging protocols.

Intravascular Ultrasound-Guided Catheter-Based Aspiration Thrombectomy in Patients With Acute Submassive Pulmonary Embolism: A Case Series.

Contrast pulmonary angiography by hand injection or power injection is widely used during catheter-based therapies for acute submassive and massive pulmonary embolism (PE). Particularly, in patients with pre-existing chronic kidney disease, this approach may present a prognostic challenge owing to a double-contrast load, initially during computed tomographic pulmonary angiography (CTPA), and during percutaneous treatment. Intravascular ultrasound (IVUS) has been used as an adjunctive imaging modality in the percutaneous treatment of chronic thromboembolic pulmonary hypertension, and in coronary and peripheral vascular interventions. We report a series of cases illustrating the use of IVUS in the management of acute PE. All five patients presented with an acute submassive PE with evidence of right ventricular (RV) strain (RV/LV ratio ≥ 0.90). Body mass index and B-type natriuretic peptide ranged from 18 to 47 kgm/m2 and 56-932 pg/mL (ref. ≤ 78), respectively. Three of the five patients had renal impairment prior to the procedure (acute kidney injury, AKI, and chronic kidney injury, CKD). Post-catheter-directed pulmonary embolectomy there was a modest reduction in mean pulmonary artery pressure in all five patients (range: -4 mmHg to -9 mmHg). The first case serves as a proof of concept of IVUS use in acute PE. This case series demonstrates that an IVUS-only approach in the catheter-directed management of acute submassive PE is feasible and may be of particular importance in patients with pre-existing renal dysfunction.

A Clinico-trichological Study of Female Androgenetic Alopecia.

BACKGROUND: Female Androgenetic Alopecia (FAGA) is one of the most common cause of diffuse scalp hair loss in women. FAGA is insidious in onset, gradually progressive, non scarring alopecia. Trichoscopy is a newly developed non invasive, objective, bed side analytical method of hair imaging based on video dermoscopy of scalp. The objective of this study is to study the clinical features of female androgenetic alopecia, to study the trichoscopic features in patients of female androgenetic alopecia and to correlate the trichoscopic features according to clinical grade of female androgenetic alopecia. METHODS: Study enrolled 50 females with FAGA and 50 normal females. FAGA cases were divided into subgroups according to sinclair scale. Laboratory investigations including hemoglobin level, triiodothyronine (T3), thyroxine (T4) thyroid stimulating hormone (TSH) and serum ferritin level were carried out in both cases and controls. After clinical diagnosis, trichoscopic examination was done and categorised using Diagnostic Criteria set by Rakowska A. et al and trichoscopic findings were coorelated with clinical grades. RESULTS: >4 yellow dots in frontal area , >2:1 ratio of single hair units (frontal: occiput) & >3:1 ratio of hair with perifollicular discoloration (frontal: occiput) are mainly seen in late stages of FAGA i.e. grade 4 & 5 and >1.5:1 ratio of vellus hairs (frontal :occiput) in early stages i.e. grade 2 & 3 while lower mean hair thickness in frontal area & > 10% thin hairs in frontal area are seen in all stages of FAGA Conclusion: Trichoscopy not only confirms the diagnosis by assessing the trichoscopic features of FAGA but also coorelates them with clinical severity. And as the age increases, grade of clinical severity increases. Not necessarily hemoglobin, thyroid and serum ferritin levels will be dearranged in FAGA cases. LIMITATION: The study is done on few number of patients. Further studies needs to be done to validate results.

High-Mobility Free-Standing InSb Nanoflags Grown on InP Nanowire Stems for Quantum Devices.

High-quality heteroepitaxial two-dimensional (2D) InSb layers are very difficult to realize because of the large lattice mismatch with other widespread semiconductor substrates. A way around this problem is to grow free-standing 2D InSb nanostructures on nanowire (NW) stems, thanks to the capability of NWs to efficiently relax elastic strain along the sidewalls when lattice-mismatched semiconductor systems are integrated. In this work, we optimize the morphology of free-standing 2D InSb nanoflags (NFs). In particular, robust NW stems, optimized growth parameters, and the use of reflection high-energy electron diffraction (RHEED) to precisely orient the substrate for preferential growth are implemented to increase the lateral size of the 2D InSb NFs. Transmission electron microscopy (TEM) analysis of these NFs reveals defect-free zinc blend crystal structure, stoichiometric composition, and relaxed lattice parameters. The resulting NFs are large enough to fabricate Hall-bar contacts with suitable length-to-width ratio enabling precise electrical characterization. An electron mobility of ∼29 500 cm2/(V s) is measured, which is the highest value reported for free-standing 2D InSb nanostructures in literature. We envision the use of 2D InSb NFs for fabrication of advanced quantum devices.

Catheter directed embolectomy of right atrial clot in transit-A case series.

In the previous literature, the prevalence of right atrial (RA) clot-in-transit associated with pulmonary embolism is around 4-18% with an associated mortality of 80-100% in untreated cases. Surgical thrombo-embolectomy has been the mainstay of treatment for stable patients but the data for percutaneous thrombo-embolectomy is lacking in the literature. We present a series of three cases of right atrium clot-in-transit treated with catheter-based therapies with Inari FlowTriever® (Inari Medical, Irvine, CA). Our three patients had different clinical profiles and presentations of right atrial clot-in-transit. All of the subjects had contraindications to surgical thrombo-embolectomy and thrombolytic therapy. Catheter based embolectomy using Inari FlowTriever® was successfully performed in all the patients. As the data on this intervention is sparse, our case series highlights successful catheter based thrombo-embolectomies in high-risk individuals with right atrial clot in-transit with or without pulmonary embolism.

Morphology control of single-crystal InSb nanostructures by tuning the growth parameters.

Research interest in indium antimonide (InSb) has increased significantly in recent years owing to its intrinsic properties and the consequent opportunities to implement next-generation quantum devices. Hence, the precise, reproducible control over morphology and crystalline quality becomes of paramount importance for a practical quantum-device technology. Here, we investigate the growth of InSb nanostructures with different morphologies on InAs stems without pre-growth efforts (patterning). InSb nanostructures such as nanowires (1D), nanoflags (2D) and nanocubes (3D) have been realized by means of Au-assisted chemical beam epitaxy by tailoring the growth parameters like growth temperature, precursor fluxes, sample rotation and substrate orientation. Through morphological and crystallographic characterization, all the as-grown InSb 2D nanostructures are found to be single-crystalline with zinc blende structure, free from any defects such as stacking faults and twin planes. The existence of two families of 2D nanostructures, characterised by an aperture angle at the base of 145° and 160°, is observed and modelled. This study provides useful guidelines for the controlled growth of high-quality InSb nanostructures with different shape.

Successful Derivation of an Induced Pluripotent Stem Cell Line from a Genetically Nonpermissive Enhanced Green Fluorescent Protein-Transgenic FVB/N Mouse Strain.

The embryonic stem cell line derivation from nonpermissive mouse strains is a challenging and highly inefficient process. The cellular reprogramming strategy provides an alternative route for generating pluripotent stem cell (PSC) lines from such strains. In this study, we successfully derived an enhanced green fluorescent protein (EGFP)-transgenic "N9" induced pluripotent stem cell (iPS cell, iPSC) line from the FVB/N strain-derived mouse embryonic fibroblasts (MEFs). The exposure of MEFs to human OCT4, SOX2, KLF4, and c-MYC (OSKM) transgenes via lentiviral transduction resulted in complete reprogramming. The N9 iPS cell line demonstrated all the criteria of a typical mouse PSC line, including normal colony morphology and karyotype (40,XY), high replication and propagation efficiencies, expression of the pluripotency-associated genes, spontaneous differentiation to three germ lineage-derived cell types, and robust potential of chimeric blastocyst formation. Taken together, using human OSKM genes for transduction, we report, for the first time, the successful derivation of an EGFP-expressing iPS cell line from a genetically nonpermissive transgenic FVB/N mouse. This cell line could provide opportunities for designing protocols for efficient derivation of PSC lines from other nonpermissive strains and developing mouse models of human diseases.

Cell culture conditions affect the ability of high content imaging assay to detect drug-induced changes in cellular parameters in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs).

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are widely used for drug safety and efficacy testing with various techniques, including high content imaging (HCI). Upon drug treatment, a significant number of hiPSC-CMs grown in regular 96-well plates coated with fibronectin detached from the bottom of the plate, complicating data acquisition. Several cell culture configurations were tested to improve cell adherence, and the effects of these configurations on total cell number, separation of feature values between the negative (DMSO 0.1%) and positive (antimycin, staurosporine) controls, scale of feature value differences, and data variability were statistically calculated. hiPSC-CMs were plated on fibronectin- (in "blanket" configuration) or MaxGel- (in "sandwich" configuration) coated plates and covered with a layer of either HydroMatrix or MaxGel 2, 7, or 11d after plating. After a total of 14d in culture, cells were treated with compounds, labeled with four fluorescent dyes (Hoechst, TMRM, NucView, and RedDot), and imaged with GE INCell2000. Based on the statistical parameters calculated, the MaxGel 25% 7d "sandwich" was superior to all other tested conditions when the cells were treated with 0.3 µM antimycin for 2 h and test compounds 10 µM crizotinib and 30 µM amiodarone for 48 h. For staurosporine treatment, the best culturing condition varied between MaxGel "sandwich" systems, depending on which parameters were under consideration. Thus, cell culturing conditions can significantly affect the ability of high content imaging to detect changes in cellular features during compound treatment and should be thoroughly evaluated before committing to compound testing.

Directed differentiation of mouse P19 embryonal carcinoma cells to neural cells in a serum- and retinoic acid-free culture medium.

P19 embryonal carcinoma cells (EC-cells) provide a simple and robust culture system for studying neural development. Most protocols developed so far for directing neural differentiation of P19 cells depend on the use of culture medium supplemented with retinoic acid (RA) and serum, which has an undefined composition. Hence, such protocols are not suitable for many molecular studies. In this study, we achieved neural differentiation of P19 cells in a serum- and RA-free culture medium by employing the knockout serum replacement (KSR) supplement. In the KSR-containing medium, P19 cells underwent predominant differentiation into neural lineage and by day 12 of culture, neural cells were present in 100% of P19-derived embryoid bodies (EBs). This was consistently accompanied by the increased expression of various neural lineage-associated markers during the course of differentiation. P19-derived neural cells comprised of NES+ neural progenitors (~ 46%), TUBB3+ immature neurons (~ 6%), MAP2+ mature neurons (~ 2%), and GFAP+ astrocytes (~ 50%). A heterogeneous neuronal population consisting of glutamatergic, GABAergic, serotonergic, and dopaminergic neurons was generated. Taken together, our study shows that the KSR medium is suitable for the differentiation of P19 cells to neural lineage without requiring additional (serum and RA) supplements. This stem cell differentiation system could be utilized for gaining mechanistic insights into neural differentiation and for identifying potential neuroactive compounds.

Aorto-Atrial Fistulas: A Contemporary Review.

Aorto-atrial fistulas (AAFs) are a relatively rare, but potentially life-threatening condition, where an anomalous connection forms between the aortic structures and the cardiac atria. AAFs are most often the result of an underlying condition concerning the cardiac structures. It may be congenital, secondary to conditions such as aortic dissection, infective endocarditis, or valve replacement, or iatrogenic in nature. Secondary causes incite local deterioration of cardiac wall integrity leading to formation of fistulous connections, whereas iatrogenic causes are more traumatic in nature. Signs and symptoms include those of volume overload and heart failure, with patients often presenting with fever, regurgitative murmurs, cardiac chamber dilation, and pedal edema. The diagnosis of AAFs requires a high degree of clinical suspicion necessitating imaging techniques such as echocardiography, computer tomography, and more invasive procedures. Management is dependent on underlying conditions and include the use of antibiotics, percutaneous closure, and in many cases, open heart surgery. It is important for physicians to be aware of this pathological condition to aid in timely management and favorable outcomes. This review attempts to summarize the various causes and clinical presentations of AAFs over the past decades.

Efficient neural differentiation of mouse pluripotent stem cells in a serum-free medium and development of a novel strategy for enrichment of neural cells.

Pluripotent stem cells (PSCs) offer an excellent model to study neural development and function. Although various protocols have been developed to direct the differentiation of PSCs into desired neural cell types, many of them suffer from limitations including low efficiency, long duration of culture, and the use of expensive, labile, and undefined growth supplements. In this study, we achieved efficient differentiation of mouse PSCs to neural lineage, in the absence of exogenous molecules, by employing a serum-free culture medium containing knockout serum replacement (KSR). Embryoid bodies (EBs) cultured in this medium predominantly produced neural cells which included neural progenitors (15-18%), immature neurons (8-24%), mature neurons (10-26%), astrocytes (27-61%), and oligodendrocytes (∼1%). Different neuronal subtypes including glutamatergic, GABAergic, cholinergic, serotonergic, and dopaminergic neurons were generated. Importantly, neurons generated in the KSR medium were electrically active. Further, the EB scooping strategy, involving the removal of the EB core region from the peripheral EB outgrowth, resulted in the enrichment of PSC-derived neural cells. Taken together, this study provides the evidence that the KSR medium is ideal for the rapid and efficient generation of neural cells, including functional neurons, from PSCs without the requirement of any other additional molecule.