Clinical and genetic investigation of 14 families with various forms of short stature syndromes.

Skeletal dysplasias are a heterogeneous group of disorders presenting mild to lethal defects. Several factors, such as genetic, prenatal, and postnatal environmental may contribute to reduced growth. Fourteen families of Pakistani origin, presenting the syndromic form of short stature either in the autosomal recessive or autosomal dominant manner were clinically and genetically investigated to uncover the underlying genetic etiology. Homozygosity mapping, whole exome sequencing, and Sanger sequencing were used to search for the disease-causing gene variants. In total, we have identified 13 sequence variants in 10 different genes. The variants in the HSPG2 and XRCC4 genes were not reported previously in the Pakistani population. This study will expand the mutation spectrum of the identified genes and will help in improved diagnosis of the syndromic form of short stature in the local population.

Biochemical properties of sheep colostrum and its potential benefits for lamb survival: a review.

Colostrum is the initial secretion of the mammary glands following parturition, which offers main food, protection, and biological active substances for the new born. The most threatening episode of neonate's life is the initial two weeks after birth. This period is associated with high neonatal mortality and morbidity. These worthwhile losses lead to a poor prolificacy rate, low profitability, and ultimately poor performance in animal production. Hence, both diseases and mortality cause valuable losses in terms of production and economic losses. The survival of neonate is correlated with their immune status and passive immune transfer (PIT). Colostrum provides the primary source of nutrition and immunity (PIT) that protects neonates against infections. It must be given as soon as possible after birth since its immunoglobulins are absorbed within the first 16-27 hours after birth, ideally within 2-4 hours. As a result, immunoglobulin (PIT) is the most important component of distressing infectious immunity, and a passable concentration of immunoglobulin in the blood of newborn lambs is linked to their health and survival rate. In this review, we summarized the importance of colostrum in early life and its association with neonatal lamb's survival, profitability and productivity of sheep farming.

Prevalence of Obesity and its Effects in Patients With COVID-19: A Systematic Review and Meta-analysis.

Background: Coronavirus disease 2019 (COVID-19) is an emerging infectious disease worldwide. Obesity has been proven to increase the susceptibility of an individual to infections, but the relationship between obesity and COVID-19 is still unclear. This study aimed to conduct a systematic review and meta-analysis of the prevalence of obesity and its effects in patients with COVID-19. Methods: Web of Science, PubMed and Embase were searched for English language studies up to May 22, 2020. We used a random or fixed-effects model to calculate pooled prevalence rates and odds ratio (OR) with 95% confidence intervals (CI). Results: Twelve studies with a total of 14 364 patients met the inclusion criteria. The pooled prevalence of obesity in patients with COVID-19 was 32.0% (95% CI, 26%-38%, P < .001). The prevalence of obesity in ICU COVID-19 patients were 37.0% (95% CI, 29%-46%, P < .001). Comparing between obese and non-obese patients, the meta-analysis showed that obesity was an important risk factor associated with COVID-19 patients needed for ICU care (OR: 1.36, 95% CI 1.22-1.52, P < .001). Conclusion: Obesity was highly prevalent (32.0%) in patients with COVID-19, especially in ICU patients (37.0%), and was an important risk factor for COVID-19 patients needed for ICU care.

Serum biochemical reference interval determination in wild Siberian weasel (Mustela sibirica).

Determining reference intervals (RI) is a valuable asset for assessing the health of wildlife species. This is the first study to establish serum biochemical RIs in Siberian weasels. Forty-two healthy free-ranging Siberian weasels were captured live and brought to Seoul Wildlife Center between June 2021 and August 2022. Blood samples from 42 healthy Siberian weasels of both sexes were used to calculate RIs. An automated analyser was used to perform serum biochemistry profiles. The American Society for Veterinary Clinical Pathology recommendations were used to calculate a nonparametric RI with 90% confidence intervals. The RIs of albumin, total protein, globulin, calcium, glucose, blood urea nitrogen, phosphorus, amylase, cholesterol, alanine aminotransferase, total bilirubin, alkaline phosphatase, creatinine, and creatine kinase were determined. The RIs established in this study will serve as a good starting point for analysing serum biochemical data in Siberian weasels.

VERTEBRAL HEART SCALE SYSTEM IN CLINICALLY HEALTHY SIBERIAN WEASEL (MUSTELA SIBIRICA).

In several species, the vertebral heart scale (VHS) measurement is frequently used to provide a more objective evaluation of cardiomegaly. However, normal-sized parameters for radiographic measures of the cardiac silhouette in Siberian weasels (Mustela sibirica) have not been determined. Right lateral (n = 24) and ventrodorsal (n = 20) thoracic radiographs from free-ranging Siberian weasels with clinically normal cardiac and pulmonary function were acquired and evaluated to determine the specific VHS for the Siberian weasel. The mean (SD) VHS of the right lateral (RL) and the ventrodorsal (VD) views were 6.70 (0.60) and 6.95 (0.69), respectively. VD view radiographs had a considerably higher VHS than RL view radiographs, and RL view measures were less variable than VD view values. The VHS of Siberian weasels was unaffected by age, body weight, or sex. The results determined in this study can be used in clinical diagnostic and radiographic evaluations of Siberian weasels.

Predictors of permanent pacemaker insertion after mitral valve replacement: A systematic review.

As the established surgical mitral valve replacement (MVR) expands toward various contemporary techniques and access routes, the predictors and burden of procedure-related complications including the need for permanent pacemaker (PPM) implantation need to be identified. Digital databases were searched systematically to identify studies reporting the incidence of PPM implantation after MVR. Detailed study and patient-level baseline characteristics including the type of study, sample size, follow-up, number of post-MVR PPM implantations, age, gender, and baseline ECG abnormalities were abstracted. A total of 12 studies, recruiting 37,124 patients were included in the final analysis. Overall, 2820 (7.6%) patients required a PPM with the net rate ranging from 1.7% to 10.96%. Post-MVR atrioventricular (AV) block was the most commonly observed indication for PPM, followed by sinoatrial (SA) node dysfunction, and bradycardia. Age, male gender, pre-existing comorbid conditions, prior CABG, history of arrhythmias or using antiarrhythmic drugs, atrial fibrillation ablation, and double valve replacement were predictors of PPM implantation post-MVR. Age, male gender, comorbid conditions like diabetes and renal impairment, prior CABG, double valve replacement, and antiarrhythmic drugs served as positive predictors of PPM implantation in patients undergoing MVR.

Evaluation of Antitermite Properties of Wood Extracts from Pongamia pinnata (L.) Pierre (Leguminosae) against Subterranean Termites.

Termiticide, repellent and antifeedant activities of extracts from Pongamia pinnata wood were evaluated against Coptotermes heimi (Wasmann) at three different concentrations preceded by a preliminary choice and no-choice tests for natural resistance of tested wood. Termites' mortality was determined in each case of extract and solvent treated Whatman filter paper. Finally, wooden blocks of poplar (19×19×19 mm) were treated with extracts and respective solvents and exposed to termites in the field for 28 days. Minimum mean weight loss was observed in dried P. pinnata (6.38%), followed by fresh P. pinnata in choice tests. In no-choice tests, dried P. pinnata was comparatively resistant with a weight loss of 12.37%, followed by fresh P. pinnata and P. deltoides. In toxicity bioassay, ethyl acetate-based wood extracts caused the highest mortality (41.66%), followed by petroleum ether, hexane, and water extracts at 10 mg/ml concentration. Similarly, ethyl acetate-based extracts showed maximum repellency (100%) followed by petroleum ether extracts at 10 mg/ml and ethyl acetate at 5 mg/ml after 60 min of termite exposure. Minimum wood losses were observed in woods treated with ethyl acetate extracts compared to control and other treatments in field experiments.

Immunocastration with gene vaccine (KISS1) induces a cell-mediated immune response in ram testis: A transcriptome evaluation.

Immunocastration vaccines achieve their effects through neutralization of the endogenous hormone by the humoral antibody produced against the immunized genes, but there is little information regarding cell-mediated immune response on the gonadal function of the immunized model is available. In this study, we used ram as a model animal to identify the cellular immune response in testicular tissues of rams immunized with intranasal KISS1 gene vaccine. The immune castration model was evaluated by sexual behaviours, spermatogenesis and serum hormone profiles after the KISS1 gene immunization. Transcriptome analysis of testicular tissues was carried out to identify the expressions of protein-coding genes involved in cellular immunity. The results showed that we successfully constructed the KISS1 immune castration ram model, in which testicular growth and development, testosterone and kisspeptin-54 levels, and sexual function were suppressed in immunized rams (p < .05). Using Hiseq™ 2000 high sequencing for ram testicular, we identified 21 differentially expressed genes (DEGs) related to cellular immunity, of which, 14 genes were upregulated and seven genes were downregulated in the testis of the immunized group (p < .05). The Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment showed that these differentially expressed genes were enriched in the antigen presentation process mediated by MHC class I and the cytotoxic pathway mediated by natural killer cells. It is concluded that KISS1 gene vaccine induced the cell-mediated immune response in testicular tissue to suppress reproductive activities in rams.

New trends in immunocastration and its potential to improve animal welfare: a mini review.

Castration is frequently used to reduce aggressive behavior and improve the meat quality of animals. Traditionally, surgical and mechanical castration are used to sterilize the animals, but these approaches are associated with a high level of pain, stress, long recovery periods, and post-operative infections. Immunocastration is a new animal-friendly, painless alternative castration technique that is used to prevent undesired sexual behavior, reduce aggressive behavior, prevent unwanted pregnancy, control wildlife populations and wandering species, enhance growth performance, improve meat quality, and treat various sex hormone-dependent disorders. The mechanism of immunocastration includes the immunological block of the hypothalamic-pituitary-gonadal axis (HPG axis) which inhibits gonadotropin secretions, causes atrophy of gonadal tissues, and inhibits gametogenesis, resulting in infertility in both female and male mammals. By the mid-1990s, various immunocastration vaccines have been tested in different animal models to achieve successful castration effects. Recently, genetic immunocastration especially DNA vaccine has gained increasing attention due to its safety, being animal-friendly, and being easy to use. This review aims to evaluate the potential of traditional castration methods, as well as the current status of immunocastration vaccines, their effects, and future prospective.

Translational precision medicine: an industry perspective.

In the era of precision medicine, digital technologies and artificial intelligence, drug discovery and development face unprecedented opportunities for product and business model innovation, fundamentally changing the traditional approach of how drugs are discovered, developed and marketed. Critical to this transformation is the adoption of new technologies in the drug development process, catalyzing the transition from serendipity-driven to data-driven medicine. This paradigm shift comes with a need for both translation and precision, leading to a modern Translational Precision Medicine approach to drug discovery and development. Key components of Translational Precision Medicine are multi-omics profiling, digital biomarkers, model-based data integration, artificial intelligence, biomarker-guided trial designs and patient-centric companion diagnostics. In this review, we summarize and critically discuss the potential and challenges of Translational Precision Medicine from a cross-industry perspective.

Greig Cephalopolysyndactyly Syndrome: Phenotypic Variability Associated with Variants in Two Different Domains of GLI3.

BACKGROUND: GLI3 is a transcriptional regulator of several genes involved in mammalian skeletal development. Mutations in the pleiotropic gene GLI3 may result in different inherited disorders including Greig cephalopolysyndactyly syndrome (GCPS). GCPS is characterized by mild to severe craniofacial and limb malformations. METHODS AND RESULTS: Here, we report clinical and molecular study of 3 families with GCPS originated in different regions of Pakistan. Sanger sequencing revealed two novel variants including a frameshift [c. 3790_3791InsC, p.(Gly1236Argfs*11)] and a missense [c.1692A>G, p.(His536Arg)], and one previously reported variant [c.1965_1966delAT, p.(His627Glufs*48)] located in 2 different domains of the GLI3. CONCLUSION: This study not only expanded spectrum of the mutations in the GLI3 but also highlighted phenotypic variability in the GCPS patients. This will facilitate diagnosis and genetic counseling of families with same and related disorders in the Pakistani population.

Assessment of groundwater intrinsic vulnerability using GIS-based DRASTIC method in District Haripur, Khyber Pakhtunkhwa, Pakistan.

The stress on the freshwater resources of the planet earth has led the United Nations to add a goal regarding clean water in sustainable development goals list in order to address the global availability of clean water. The widespread use of fertilizers and industrial effluents caused the groundwater contamination in the Haripur District, Khyber Pakhtunkhwa, Pakistan. To investigate and assess the vulnerability of groundwater to contamination, geographic information system (GIS)-based DRASTIC model has been employed. The DRASTIC index values lie between 88 and 190. The lower the DI value, the lower will be the susceptibility towards pollution and vice versa. The indices were classified into five zones, i.e., low (< 109), medium (110-129), moderate (130-149), high (150-169), and very high vulnerable zones (> 170) on the basis of equal intervals. The low vulnerable zone covers almost 6% of the study area, i.e., 118 km2. Medium vulnerable zone encompasses an area of approximately 23%, i.e., 506 km2. The moderate vulnerable zone is the largest in the district covering almost 965 km2, approximately 45% of the study area. The high and very high vulnerable zone encompasses almost 23% and 3% of the study area which means 506 km2 and 66 km2 respectively. A spatial distribution map was generated for nitrate concentration to validate the DRASTIC indices. The results demonstrate a fair relation between groundwater susceptibility and spatial nitrate distribution. This index map will provide a baseline study for this area to develop the safe zones for groundwater exploitation and controlling the current state of deterioration of environmental norms. The areas of high vulnerability are the firsthand task to improve the current situation of crisis especially in the southern parts such as the Hattar industrial area and its surroundings drained by those effluents. These further need specific tasks to restore and reclaim the polluted ecosystem by using proper technological solutions for disposal of these effluents.

Multipathway synergy promotes testicular transition from growth to spermatogenesis in early-puberty goats.

BACKGROUND: The microscopic process of postnatal testicular development in early-puberty animals is poorly understood. Therefore, in this study, 21 male Yiling goats with average ages of 0, 30, 60, 90, 120, 150 and 180 days old (each age group comprised three goats) were used to study the changes in organs, tissues and transcriptomes during postnatal testicle development to obtain a broad and deep insight into the dynamic process of testicular transition from growth to spermatogenesis in early-puberty animals. RESULTS: The inflection point of testicular weight was at 119 days postpartum (dpp), and the testicular weight increased rapidly from 119 dpp to 150 dpp. Spermatozoa were observed in the testis at 90 dpp by using haematoxylin-eosin staining. We found from the transcriptome analysis of testes that the testicular development of Yiling goat from birth to 180 dpp experienced three stages, namely, growth, transition and spermatogenesis stages. The goats in the testicular growth stage (0-60 dpp) showed a high expression of growth-related genes in neurogenesis, angiogenesis and cell junction, and a low expression of spermatogenesis-related genes. The goats aged 60-120 dpp were in the transitional stage which had a gradually decreased growth-related gene transcription levels and increased spermatogenesis-related gene transcription levels. The goats aged 120-180 dpp were in the spermatogenesis stage. At this stage, highly expressed spermatogenesis-related genes, downregulated testicular growth- and immune-related genes and a shift in the focus of testicular development into spermatogenesis were observed. Additionally, we found several novel hub genes, which may play key roles in spermatogenesis, androgen synthesis and secretion, angiogenesis, cell junction and neurogenesis. Moreover, the results of this study were compared with previous studies on goat or other species, and some gene expression patterns shared in early-puberty mammals were discovered. CONCLUSIONS: The postnatal development of the testis undergoes a process of transition from organ growth to spermatogenesis. During this process, spermatogenesis-related genes are upregulated, whereas neurogenesis-, angiogenesis-, cell junction-, muscle- and immune-related genes are downregulated. In conclusion, the multipathway synergy promotes testicular transition from growth to spermatogenesis in early-puberty goats and may be a common rule shared by mammals.

High Coercivity and Magnetization in WSe2 by Codoping Co and Nb.

Introducing ferromagnetism in transition metal dichalcogenides has attracted lots of attention due to the possible applications in spintronics devices. Generally, single magnetic element doping is used to introduce magnetism. However, mostly, weak ferromagnetism is observed. In this work, codoping of two kinds of transition metals (Nb and Co) into WSe2 is used to study its magnetic properties. In detail, single crystal WSe2 is codoped with 4 at% Co and various concentrations of Nb by employing the physical ion implantation method. Raman, X-ray diffraction and X-ray photoelectron spectroscopy results reveal the effective substitutional doping of implanted elements (Co and Nb). Magnetic measurements illustrate that both un-doped and 4 at% Co doped WSe2 show weak ferromagnetism whereas magnetization is strongly enhanced when Co and Nb are codoped into WSe2 . The magnetization is comparable with a ferromagnet, which may be attributed to Co, Nb doping and defects. In addition, a large coercivity of ≈1.2 kOe is observed in the 1 at% Nb-4 at% Co codoped WSe2 sample, which may be ascribed to the combined effect of doping-induced stress, defect-dictated pinning and anisotropy of NbSe bond owing to the charge transfer between Nb and Se ions.

Genetic engineering and bacterial pathogenesis against the vectorial capacity of mosquitoes.

Mosquitoes are the main vector of multiple diseases worldwide and transmit viral (malaria, chikungunya, encephalitis, yellow fever, as well as dengue fever), as well as bacterial diseases (tularemia). To manage the outbreak of mosquito populations, various management programs include the application of chemicals, followed by biological and genetic control. Here we aimed to focus on the role of bacterial pathogenesis and molecular tactics for the management of mosquitoes and their vectorial capacity. Bacterial pathogenesis and molecular manipulations have a substantial impact on the biology of mosquitoes, and both strategies change the gene expression and regulation of disease vectors. The strategy for genetic modification is also proved to be excellent for the management of mosquitoes, which halt the development of population via incompatibility of different sex. Therefore, the purpose of the present discussion is to illustrate the impact of both approaches against the vectorial capacity of mosquitoes. Moreover, it could be helpful to understand the relationship of insect-pathogen and to manage various insect vectors as well as diseases.

Magnetic resonance imaging reveals specific anatomical changes in the brain of Agat- and Gamt-mice attributed to creatine depletion and guanidinoacetate alteration.

Arginine:glycine amidinotransferase- and guanidinoacetate methyltransferase deficiency are severe neurodevelopmental disorders. It is not known whether mouse models of disease express a neuroanatomical phenotype. High-resolution magnetic resonance imaging (MRI) with advanced image analysis was performed in perfused, fixed mouse brains encapsulated with the skull from male, 10-12 week old Agat -exc and B6J.Cg-Gamt tm1Isb mice (n = 48; n = 8 per genotype, strain). T2-weighted MRI scans were nonlinearly aligned to a 3D atlas of the mouse brain with 62 structures identified. Local differences in brain shape related to genotype were assessed by analysis of deformation fields. Creatine (Cr) and guanidinoacetate (GAA) were measured with high-performance liquid chromatography (HPLC) in brain homogenates (n = 24; n = 4 per genotype, strain) after whole-body perfusion. Cr was decreased in the brain of Agat- and Gamt mutant mice. GAA was decreased in Agat-/- and increased in Gamt-/- . Body weight and brain volume were lower in Agat-/- than in Gamt-/- . The analysis of entire brain structures revealed corpus callosum, internal capsule, fimbria and hypothalamus being different between the genotypes in both strains. Eighteen and fourteen significant peaks (local areas of difference in relative size) were found in Agat- and Gamt mutants, respectively. Comparing Agat-/- with Gamt-/- , we found changes in three brain regions, lateral septum, amygdala, and medulla. Intra-strain differences in four brain structures can be associated with Cr deficiency, while the inter-strain differences in three brain structures of the mutant mice may relate to GAA. Correlating these neuroanatomical findings with gene expression data implies the role of Cr metabolism in the developing brain and the importance of early intervention in patients with Cr deficiency syndromes.

Ophthalmic findings in cinereous vultures (Aegypius monachus).

OBJECTIVE: The aim of the present study was to provide ophthalmic reference values under normal physiological conditions for Aegypius monachus (cinereous vulture). PROCEDURES: Thirty-two eyes of sixteen adult captive cinereous vultures were used for this study. Tear tests and tonometry in conscious and anesthetized states, neuro-ophthalmic tests, measurement of corneal diameter, slit-lamp biomicroscopy, ophthalmoscopy, and funduscopy were performed. RESULTS: Schirmer tear test (STT) value was 11.4 ± 2.6 and 11.5 ± 2.8 mm/min in the right (OD) and left eye (OS), respectively. Phenol red thread test (PRT) values were 22.3 ± 2.1 mm/15 s OD and 22.8 ± 3.0 mm/15 s OS. The results showed a strong correlation between STT and PRT in both eyes. Intraocular pressure (IOP) values were 32.8 ± 6.9 mm Hg OD and 31.9 ± 7.1 mm Hg OS with TonoVet and 20.7 ± 4.5 mm Hg OD and 19.5 ± 4.1 mm Hg OS with Tono-Pen. There were significant differences in IOPs between rebound and applanation tonometry in both OD and OS. Tear production and IOP values showed significant reductions with general anesthesia in both tear tests and both tonometry (P < .001). Horizontal corneal diameter (mm) was 15.56 ± 0.96 OD and 15.56 ± 0.96 OS. Vertical diameter (mm) was 14.13 ± 0.96 OD and 14.06 ± 1.06 OS. The horizontal diameter was significantly longer than vertical diameter (P < .001). CONCLUSIONS: Ocular morphologic information and normal reference range values for various ophthalmic measurements were obtained in clinically healthy cinereous vultures, which can facilitate accurate diagnosis and better management of ophthalmic diseases in cinereous vultures.

The Rho GTPase Rif signals through IRTKS, Eps8 and WAVE2 to generate dorsal membrane ruffles and filopodia.

Rif induces dorsal filopodia but the signaling pathway responsible for this has not been identified. We show here that Rif interacts with the I-BAR family protein IRTKS (also known as BAIAP2L1) through its I-BAR domain. Rif also interacts with Pinkbar (also known as BAIAP2L2) in N1E-115 mouse neuroblastoma cells. IRTKS and Rif induce dorsal membrane ruffles and filopodia. Dominant-negative Rif inhibits the formation of IRTKS-induced morphological structures, and Rif activity is blocked in IRTKS-knockout (KO) cells. To further define the Rif-IRTKS signaling pathway, we identify Eps8 and WAVE2 (also known as WASF2) as IRTKS interactors. We find that Eps8 regulates the size and number of dorsal filopodia and membrane ruffles downstream of Rif-IRTKS signaling, whereas WAVE2 modulates dorsal membrane ruffling. Furthermore, our data suggests that Tir, a protein essential for enterohemorrhagic Escherichia coli infection, might compete for Rif for interaction with the I-BAR domain of IRTKS. Based on this evidence, we propose a model in which Rho family GTPases use the I-BAR proteins, IRSp53 (also known as BAIAP2), IRTKS and Pinkbar, as a central mechanism to modulate cell morphology.

OpenSegSPIM: a user-friendly segmentation tool for SPIM data.

UNLABELLED: OpenSegSPIM is an open access and user friendly 3D automatic quantitative analysis tool for Single Plane Illumination Microscopy data. The software is designed to extract, in a user-friendly way, quantitative relevant information from SPIM image stacks, such as the number of nuclei or cells. It provides quantitative measurement (volume, sphericity, distance, intensity) on Light Sheet Fluorescent Microscopy images. AVAILABILITY AND IMPLEMENTATION: freely available from http://www.opensegspim.weebly.com Source code and binaries under BSD License. CONTACT: lgole@imcb.a-star.edu.sg or wmyu@imcb.a-star.edu.sg or sohail.ahmed@imb.a-star.edu.sg SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

NeuronCyto II: An automatic and quantitative solution for crossover neural cells in high throughput screening.

Microscopy is a fundamental technology driving new biological discoveries. Today microscopy allows a large number of images to be acquired using, for example, High Throughput Screening (HTS) and 4D imaging. It is essential to be able to interrogate these images and extract quantitative information in an automated fashion. In the context of neurobiology, it is important to automatically quantify the morphology of neurons in terms of neurite number, length, branching and complexity, etc. One major issue in quantification of neuronal morphology is the "crossover" problem where neurites cross and it is difficult to assign which neurite belongs to which cell body. In the present study, we provide a solution to the "crossover" problem, the software package NeuronCyto II. NeuronCyto II is an interactive and user-friendly software package for automatic neurite quantification. It has a well-designed graphical user interface (GUI) with only a few free parameters allowing users to optimize the software by themselves and extract relevant quantitative information routinely. Users are able to interact with the images and the numerical features through the Result Inspector. The processing of neurites without crossover was presented in our previous work. Our solution for the "crossover" problem is developed based on our recently published work with directed graph theory. Both methods are implemented in NeuronCyto II. The results show that our solution is able to significantly improve the reliability and accuracy of the neurons displaying "crossover." NeuronCyto II is freely available at the website: https://sites.google.com/site/neuroncyto/, which includes user support and where software upgrades will also be placed in the future. © 2016 The Authors. Cytometry Part A Published by Wiley Periodicals, Inc. on behalf of ISAC.