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Differential effects of integrin-linked kinase inhibitor Cpd22 on severe pulmonary hypertension in male and female rats
Pulmonary arterial hypertension (PAH) is a progressive fatal disease with no cure. Inhibition of integrin-linked kinase (ILK) reverses experimental pulmonary hypertension (PH) in male mice, but its effect on severe experimental PH in either male or female animals is unknown. We examined effects of ILK inhibitor Cpd22 on rats with SU5416/hypoxia-induced PH; treatment was performed at six to eight weeks after PH initiation. Five weeks after PH initiation, male and female rats developed similar levels of PH. Eight weeks after PH induction, vehicle-treated male rats had more severe PH than females. Cpd22-treated males, but not females, showed complete suppression of phospho-Akt in small pulmonary arteries (PAs), significantly lower PA medial thickness and percentage of fully occluded arteries, decreased systolic right ventricle (RV) pressure, PA pressure, RV hypertrophy, RV end-diastolic pressure, and improved RV contractility index compared to vehicle-treated group. Cpd22 suppressed proliferation of human male and female PAH pulmonary artery vascular smooth muscle cell (PAVSMC). 17β-estradiol had no effect as a single agent but significantly attenuated Cpd22-dependent inhibition of proliferation in female, but not male, PAH PAVSMC. Taken together, these data demonstrate that male rats develop more severe PH than females but respond better to Cpd22 treatment by reducing pulmonary vascular remodeling, PH, and RV hypertrophy and improving RV functional outcomes. 17β-estradiol diminishes anti-proliferative effect of Cpd22 in female, but not male, human PAH PAVSMC. These findings suggest potential attractiveness of ILK inhibition to reduce established PH in males and suggest that the combination with estrogen-lowering drugs could be considered to maximize anti-proliferative and anti-remodeling effects of ILK inhibitors in females.
Journal Article
Flow cytometry of hematological malignancies
Flow Cytometry of Hematological Malignancies contains an array of graphical outputs produced by the technique in the study of the most (and the least) common diseases. The images included allow you to compare your own results with a third party reference pattern. There is a detailed description of the main leukocyte antigens, together with a description of their distribution amongst normal and abnormal blood cells. The book also provides a comprehensive description of the phenotype of every neoplastic blood disease recorded in the WHO classification system, including all the instructions needed to recognise and classify even the least common entity. Designed to be practical, the book is perfect for quick consultation and is divided into two main sections. Section I deals with the direct object of immunophenotyping, and Section II deals with the ultimate target of the analysis. More than 50 antigens are covered and every antigen is dealt with in three main parts: general features, cytometric features and practical hints. This authoritative and state-of-the-art reference will be invaluable for clinicians directly involved in the diagnosis and analysis of hematological diseases, including hematologists, hematopathologists, oncologists, pathologists and technicians working in diagnostic laboratories.
eBook
Guide to research techniques in neuroscience
Neuroscience is, by definition, a multidisciplinary field: some scientists study genes and proteins at the molecular level while others study neural circuitry using electrophysiology and high-resolution optics.A single topic can be studied using techniques from genetics, imaging, biochemistry, or electrophysiology.
eBook
Practical medical microbiology for clinicians
Infectious diseases constitute a major portion of illnesses worldwide, and microbiology is a main pillar of clinical infectious disease practice. Knowledge of viruses, bacteria, fungi, and parasites is integral to practice in clinical infectious disease.
Practical Medical Microbiology is an invaluable reference for medical microbiology instructors. Drs. Berkowitz and Jerris are experienced teachers in the fields of infectious diseases and microbiology respectively, and provide expert insight into microorganisms that affect patients, how organisms are related to each other, and how they are isolated and identified in the microbiology laboratory. The text also is designed to provide clinicians the knowledge they need to facilitate communication with the microbiologist in their laboratory.
The text takes a systematic approach to medical microbiology, describing taxonomy of human pathogens and consideration of organisms within specific taxonomic groups. The text tackles main clinical infections caused by different organisms, and supplements these descriptions with clinical case studies, in order to demonstrate the effects of various organisms.
Practical Medical Microbiology is an invaluable resource for students, teachers, and researchers studying clinical microbiology, medical microbiology, infectious diseases, and virology.
eBook
The H2 robotic exoskeleton for gait rehabilitation after stroke: early findings from a clinical study
Background
Stroke significantly affects thousands of individuals annually, leading to considerable physical impairment and functional disability. Gait is one of the most important activities of daily living affected in stroke survivors. Recent technological developments in powered robotics exoskeletons can create powerful adjunctive tools for rehabilitation and potentially accelerate functional recovery. Here, we present the development and evaluation of a novel lower limb robotic exoskeleton, namely H2 (Technaid S.L., Spain), for gait rehabilitation in stroke survivors.
Methods
H2 has six actuated joints and is designed to allow intensive overground gait training. An assistive gait control algorithm was developed to create a force field along a desired trajectory, only applying torque when patients deviate from the prescribed movement pattern. The device was evaluated in 3 hemiparetic stroke patients across 4 weeks of training per individual (approximately 12 sessions). The study was approved by the Institutional Review Board at the University of Houston. The main objective of this initial pre-clinical study was to evaluate the safety and usability of the exoskeleton. A Likert scale was used to measure patient’s perception about the easy of use of the device.
Results
Three stroke patients completed the study. The training was well tolerated and no adverse events occurred. Early findings demonstrate that H2 appears to be safe and easy to use in the participants of this study. The overground training environment employed as a means to enhance active patient engagement proved to be challenging and exciting for patients. These results are promising and encourage future rehabilitation training with a larger cohort of patients.
Conclusions
The developed exoskeleton enables longitudinal overground training of walking in hemiparetic patients after stroke. The system is robust and safe when applied to assist a stroke patient performing an overground walking task. Such device opens the opportunity to study means to optimize a rehabilitation treatment that can be customized for individuals.
Trial registration:
This study was registered at ClinicalTrials.gov (
https://clinicaltrials.gov/show/NCT02114450
).
Journal Article
Implementation of Nanopore sequencing as a pragmatic workflow for copy number variant confirmation in the clinic
Background
Diagnosis of rare genetic diseases can be a long, expensive and complex process, involving an array of tests in the hope of obtaining an actionable result. Long-read sequencing platforms offer the opportunity to make definitive molecular diagnoses using a single assay capable of detecting variants, characterizing methylation patterns, resolving complex rearrangements, and assigning findings to long-range haplotypes. Here, we demonstrate the clinical utility of Nanopore long-read sequencing by validating a confirmatory test for copy number variants (CNVs) in neurodevelopmental disorders and illustrate the broader applications of this platform to assess genomic features with significant clinical implications.
Methods
We used adaptive sampling on the Oxford Nanopore platform to sequence 25 genomic DNA samples and 5 blood samples collected from patients with known or false-positive copy number changes originally detected using short-read sequencing. Across the 30 samples (a total of 50 with replicates), we assayed 35 known unique CNVs (a total of 55 with replicates) and one false-positive CNV, ranging in size from 40 kb to 155 Mb, and assessed the presence or absence of suspected CNVs using normalized read depth.
Results
Across 50 samples (including replicates) sequenced on individual MinION flow cells, we achieved an average on-target mean depth of 9.5X and an average on-target read length of 4805 bp. Using a custom read depth-based analysis, we successfully confirmed the presence of all 55 known CNVs (including replicates) and the absence of one false-positive CNV. Using the same CNV-targeted data, we compared genotypes of single nucleotide variant loci to verify that no sample mix-ups occurred between assays. For one case, we also used methylation detection and phasing to investigate the parental origin of a 15q11.2-q13 duplication with implications for clinical prognosis.
Conclusions
We present an assay that efficiently targets genomic regions to confirm clinically relevant CNVs with a concordance rate of 100%. Furthermore, we demonstrate how integration of genotype, methylation, and phasing data from the Nanopore sequencing platform can potentially simplify and shorten the diagnostic odyssey.
Journal Article