Functional role of ompF and ompC porins in pathogenesis of avian pathogenic Escherichia coli.
Avian pathogenic Escherichia coli is an important pathogen causes systemic infections in avian species and large economic losses in poultry industry worldwide. The functional role of porins during the infection and their mechanisms of interaction with host tissues for adhesion to and invasion are poorly understood. However, whether porins play a role in infection remains unclear. In this study we evaluated the potential of ompF and ompC outer membrane porins in the pathogenesis of avian pathogenic E. coli (APEC) strain TW-XM. The ompF and ompC were deleted to generate a series of mutants. We found that, ΔompF and ΔompC reduced significantly the adherence by 41.3% and 46.1% and invasion capabilities of APEC to mouse brain microvascular endothelial cell (BMEC) bEnd.3 cells in vitro by 51.9% and 49.7% respectively, compared with the wild strain TW-XM. In vivo experiment based on the measurement of the LD50 have also shown that, ΔompF and ΔompC reduced the bacterial virulence by 9.8-fold, 12.3-fold in ducklings and 9-fold, 10.2-fold in mouse models. Animal infection experiments further revealed that, loss of ompF and ompC reduced TW-XM colonization and invasion capacity in brains, lungs and blood compared to wild-type strain TW-XM (P > 0.01). These virulence-related phenotypes were partially recoverable by genetic complementation. The results of the quantitative real-time reverse transcription-PCR (qRT-PCR) indicated that, the loss of ompF and ompC significantly decreased the expression levels of ompA, fimC and iBeA genes in the mutant strains, compared to wild-type strainTW-XM (P < 0.01). Collectively, our data demonstrate that inactivation of these two porins decreased adhesion, invasion, colonization, proliferation capacities, possibly by reduced expression levels of ompA, fimC and iBeA, which may indicate the involvement of ompF and ompC in APEC pathogenesis.


Probabilistic finite element prediction of knee wear simulator mechanics.
Computational models have recently been developed to replicate experimental conditions present in the Stanmore knee wear simulator. These finite element (FE) models, which provide a virtual platform to evaluate total knee replacement (TKR) mechanics, were validated through comparisons with experimental data for a specific implant. As with any experiment, a small amount of variability is inherently present in component alignment, loading, and environmental conditions, but this variability has not been previously incorporated in the computational models. The objectives of the current research were to assess the impact of experimental variability on predicted TKR mechanics by determining the potential envelope of joint kinematics and contact mechanics present during wear simulator loading, and to evaluate the sensitivity of the joint mechanics to the experimental parameters. In this study, 8 component alignment and 4 experimental parameters were represented as distributions and used with probabilistic methods to assess the response of the system, including interaction effects. The probabilistic FE model evaluated two levels of parameter variability (with standard deviations of component alignment parameters up to 0.5mm and 1 degrees ) and predicted a variability of up to 226% (3.44mm) in resulting anterior-posterior (AP) translation, up to 169% (4.30 degrees ) in internal-external (IE) rotation, but less than 10% (1.66MPa) in peak contact pressure. The critical alignment parameters were the tilt of the tibial insert and the IE rotational alignment of the femoral component. The observed variability in kinematics and, to a lesser extent, contact pressure, has the potential to impact wear observed experimentally.


Hemodynamic effects of isometric exercise and mental arithmetic in hypertension treated with selective and nonselective beta-blockade.
Hypertensive patients have an unfavorable pressor response to exogenous epinephrine during nonselective beta-blockade. We studied hemodynamics during epinephrine release induced by handgrip exercise and mental arithmetic to examine the clinical relevance of this phenomenon. Twenty-two hypertensive patients were examined in a double-blind crossover experiment with placebo, propranolol (240 mg daily), placebo-washout, and metoprolol (300 mg daily). Changes induced by stress tests for systolic and diastolic blood pressure, heart rate (HR), and forearm blood flow (FBF) were of the same order on both beta-blockers. Rises in HR and FBF were equally reduced by both drugs. Neither handgrip exercise nor mental arithmetic induced significant differences in reaction during selective and nonselective beta-blockade.


Maternal pulse oximetry perfusion index as a predictor of early adverse respiratory neonatal outcome after elective cesarean delivery.
Evidence suggests increased morbidity, in particular early neonatal respiratory complications, in newborns from elective cesarean section compared with those from vaginal delivery. No reliable maternal predictors of adverse neonatal outcome at elective cesarean section are known. Here, we prospectively tested the hypothesis that a low maternal perfusion index at the baseline phase (i.e., preanesthesia) of the elective cesarean section is a predictor of early adverse neonatal respiratory outcome. Prospective cohort study. Operating and delivery rooms of a public health hospital with a tertiary-level neonatal intensive care unit. Forty-four healthy pregnant women with no known risk factors undergoing elective cesarean section at term gestation. Elective cesarean section was divided into nine phases. Analysis of pulse oximetry-derived signals (perfusion index, pulse rate, and oximetry) and systolic, diastolic, and differential blood pressure were recorded. Maternal arterial and venous newborn cord blood gas analyses and placental histology were evaluated. Early respiratory complications (transient tachypnea of the newborn, n = 5; respiratory distress syndrome, n = 1) were observed in 13.6% (6 of 44) of the newborns. A maternal perfusion index < or = 1.9 (lower quartile) during the preanesthesia phase of the elective cesarean section was an independent predictor of early adverse neonatal respiratory outcome (odds ratio 68.0, 95% confidence interval 6.02-767.72; p < .0001). A decreased perfusion index value in the preanesthesia phase of elective cesarean section is a maternal predictor of increased neonatal morbidity and is significantly related to subclinical placental inflammatory disease. These observations suggest the feasibility of a noninvasive pulse oximeter prenatal screening of the high-risk fetus/newborn in elective cesarean section.


Thinking outside the cell: proteases regulate hepatocyte division.
The liver has the unique ability to regenerate after loss of mass and function such as following surgical resection or toxic liver injury. Gene targeting has identified factors crucial to liver development and regeneration. Regeneration occurs through growth-factor- and cytokine-mediated proliferation of differentiated hepatocytes, and extracellular proteases are now recognized to process these molecules. Proteases release cytokines and growth factors that are anchored to the hepatic extracellular matrix or require processing for their bioactivity. Crucial 'start and stop' signals for liver regeneration are regulated by serine proteases and metalloproteases that provide an interface between proteolytic cascades and intracellular signaling during hepatocyte division.


Does pelvic congestion syndrome influence symptoms of chronic venous disease of the lower extremities?
The study was aimed at assessing the influence of pelvic congestion syndrome (PCS) on the clinical manifestations of chronic venous disease (CVD) of the lower extremities in patients with concomitant varicose veins of the pelvis and lower extremities. We examined clinically and with duplex ultrasound 30 women with varicose veins of the lower extremity only (VVLE) and another 45 women with VVLE and concomitant varicose veins of the pelvis (VVP) causing symptoms of the pelvic congestion syndrome (PCS). All patients had chronic venous disease (CVD) of class C2 (n = 47) or C3 (n = 28) according to the CEAP classification. All patients underwent duplex ultrasound (DUS) of the lower extremities and pelvis. Based on the clinical examination and DUS findings, the patients were allocated into two groups: group 1 (n = 30 patients with isolated VVLE without the signs of PCS and pelvic veins lesions) and group 2 (n = 45 patients with concomitant PCS, VVP and VVLE). The rates and severity of varicose, pain, and edema syndromes, and leg heaviness were assessed. The patients of the group 2 had significantly more severe clinical manifestations of CVD, 4 times higher risk of leg pain (odds ratio [OR] 4.23; 95% CI 1.57-11.39), 7 times higher risk of leg edema (OR 7.42; 95% CI 2.23-24.78), 5 times higher risk of leg heaviness (OR 5.3; 95% CI 1.85-15.07), and in general 2 times more severe varicose veins, compared with the group 1. The PCS is associated with an increase in the incidence and severity of the CVD symptoms.


Attention and memory benefits for physical attractiveness may mediate prosocial biases.
Mating motivations can explain attractiveness benefits, but what proximate mechanisms might serve as efficient causes of these biases? There is growing evidence that visual cues of physical attractiveness capture attention and facilitate memory, enhancing salience in ways that could underlie, for example, preferring one job applicant over another. All of these effects beg deeper questions about the meaning of attractiveness.


Heterogeneity of clonal expansion and maturation-linked mutation acquisition in hematopoietic progenitors in human acute myeloid leukemia.
Recent technological advances led to an appreciation of the genetic complexity of human acute myeloid leukemia (AML), but underlying progenitor cells remain poorly understood because their rarity precludes direct study. We developed a co-culture method integrating hypoxia, aryl hydrocarbon receptor inhibition and micro-environmental support via human endothelial cells to isolate these cells. X-chromosome inactivation studies of the least mature precursors derived following prolonged culture of CD34(+)/CD33(-) cells revealed polyclonal growth in highly curable AMLs, suggesting that mutations necessary for clonal expansion were acquired in more mature progenitors. Consistently, in core-binding factor (CBF) leukemias with known complementing mutations, immature precursors derived following prolonged culture of CD34(+)/CD33(-) cells harbored neither mutation or the CBF mutation alone, whereas more mature precursors often carried both mutations. These results were in contrast to those with leukemias with poor prognosis that showed clonal dominance in the least mature precursors. These data indicate heterogeneity among progenitors in human AML that may have prognostic and therapeutic implications.


Multidimensional Proteomic Approach of Endothelial Progenitors Demonstrate Expression of KDR Restricted to CD19 Cells.
Endothelial progenitor cells (EPCs) are involved in vasculogenesis and cardiovascular diseases. However, the phenotype of circulating EPCs remains elusive but they are more often described as CD34+KDR+. The aim of the study was to extensively characterize circulating potential vasculogenic stem cell candidates in two populations of patients with cardiovascular disease by powerful multidimensional single cell complementary cytometric approaches (mass, imaging and flow). We identified cellular candidates in one patient before and after bioprosthetic total artificial heart implantation and results were confirmed in healthy peripheral and cord blood by mass cytometry. We also quantified cellular candidates in 10 patients with different COVID-19 severity. Both C-TAH implantation and COVID-19 at critical stage induce a redistribution of circulating CD34+ and CD19+ sub-populations in peripheral blood. After C-TAH implantation, circulating CD34+ progenitor cells expressed c-Kit stem marker while specific subsets CD34+CD133-/+CD45-/dimc-Kit+KDR- were mobilized. KDR was only expressed by CD19+ B-lymphocytes and CD14+ monocytes subpopulations in circulation. We confirmed by mass cytometry this KDR expression on CD19+ in healthy peripheral and cord blood, also with a VE-cadherin expression, confirming absence of endothelial lineage marker on CD34+ subtypes. In COVID-19, a significant mobilization of CD34+c-Kit+KDR- cells was observed between moderate and critical COVID-19 patients regardless CD133 or CD45 expression. In order to better evaluate EPC phenotype, we performed imaging flow cytometry measurements of immature CD34+KDR+ cells in cord blood and showed that, after elimination of non-circular events, those cells were all CD19+. During COVID-19, a significant mobilization of CD19+KDR+ per million of CD45+ cells was observed between moderate and critical COVID-19 patients regardless of CD34 expression. CD34+c-Kit+ cells are mobilized in both cardiovascular disease described here. KDR cells in peripheral blood are CD19 positive cells and are not classic vasculogenic stem and/or progenitor cells. A better evaluation of c-Kit and KDR expressing cells will lead to the redefinition of circulating endothelial progenitors.Graphical abstract Central illustration figure. Multidimensional proteomic approach of endothelial progenitors demonstrate expression of KDR restricted to CD19 cells. Endothelial progenitor cells (EPCs) are involved in cardiovascular diseases, however their phenotype remains elusive. We elucidated here EPCs phenotype by a deep characterization by multidimensional single cell complementary cytometric approaches after Bioprosthetic total artificial heart implantation and during COVID-19. We showed a redistribution of circulating CD34+ and CD19+ sub-populations in both situations. None of the immature cell population expresses KDR. Mobilized CD34+ expressed c-Kit. Imaging flow cytometry demonstrated that CD34+KDR+ cells, after elimination of non-circular events, are all CD19+. Our results suggest a new definition of circulating EPCs and emphasize involvement of CD19 cells in cardiovascular disease.


Perfluorochemicals for gas transport and improvement of cell cultures.
This paper has considered the effects and potential application of PFCs, their emulsions and emulsion components for regulating growth and metabolic functions of microbial, animal and plant cells in culture. PFCs will help to overcome problems encountered in conventional culture systems (e.g. limited gas supply, mechanical damage), especially where cells are grown to high density. While the commercial potential of PFCs for in vitro systems has not yet been fully exploited, the most exciting areas for future developments are in the culture of animal and plant cell lines of importance in biotechnology and medicine.


Use of a synthetic dural substitute to prevent ventral retethering in the management of diastematomyelia.
Diastematomyelia is a congenital condition where the spinal cord is split by a bony or cartilaginous septum. Neurological signs and symptoms arise when this septum tethers the spinal cord. Surgical detethering often improves symptoms; however, recurrent tethering of the cord is increasingly recognised as a long-term complication. In order to prevent retethering many techniques have been used, including early patient mobilisation and sectioning of the cord. Dorsal expansile duroplasty, using synthetic grafts, is a commonly used technique to prevent recurrent dorsal tethering. We present a 31-year-old woman with recurrent ventral tethering of the cord where we used expanded polytetrafluoroethylene (Gore Preclude MVP Dura Substitute; WL Gore and Associates, Flagstaff, AZ, USA) to cover the ventral dural surface, separating the cord from its dural site of tethering. This technique may be useful to prevent ventral retethering in diastematomyelia.


Outcome of 2 cc pure sulfur hexafluoride gas tamponade for macular hole surgery.
Isoexpansile concentrations of intraocular gases are typically used as tamponading agent in macular hole surgery. Using a small volume of the pure form of these gases may achieve the same result without increasing the incidence of postoperative complications. The purpose of this study was to evaluate the anatomical and visual outcomes following macular hole surgery with 2 cc pure (100 %) sulfur hexafluoride (SF6) gas tamponade. A retrospective study of eyes with idiopathic macular holes that underwent 23-gauge pars plana vitrectomy with 2 cc pure SF6 gas tamponade. Macular hole surgery was performed alone or in combination with phacoemulsification in eyes with cataract. Preoperative and postoperative data including best corrected visual acuity recorded in LogMAR units, slit-lamp biomicroscopy, and optical coherence tomography were analysed. Surgical complications were also recorded. Seventy six eyes of seventy five patients were analysed. A closure rate of 100 % was achieved with reoperation in 4 eyes. There was a significant improvement in best-corrected visual acuity from a mean of 0.65 LogMAR preoperatively to 0.36 at 6 months (p value 0.004). Forty five (59 %) eyes gained at least 2 lines on the Snellen visual acuity chart. Postoperative elevation in intraocular pressure (≥30 mmHg) was documented in 3 eyes (4 %). Macular hole surgery with 2 cc pure SF6 gas tamponade achieved a high success rate with a low incidence of complications. The smaller volume of gas required makes it a cheaper technique.


Brachial plexus injury occurring during oral and maxillofacial surgery: a case report.
A case of brachial plexus injury occurring during bilateral temporomandibular joint reconstruction is reported. Patient positioning during surgery is the key to prevention of such injuries.


NRF2 level is negatively correlated with TGF-β1-induced lung cancer motility and migration via NOX4-ROS signaling.
Transforming growth factor-β1 (TGF-β1) is a multifaceted factor in cancer biology that regulates cell proliferation and migration. Overactivation of nuclear factor erythroid 2-like 2 (NFE2L2; NRF2) in cancers has been associated with facilitated tumor growth and therapy resistance; however, role in cancer migration has not been clearly explained yet. In this study, we investigated the role of NRF2 on TGF-β1-induced cell motility/migration. In NRF2-silenced lung cancer A549 cells, both basal and TGF-β1-inducible cell motility/migration increased compared to those in A549. SMAD transcription activity and phosphorylated SMAD2/3 levels were higher in TGF-β1-treated NRF2-low A549 cells than those in A549. Notably, the levels of reactive oxygen species (ROS) that were elevated by TGF-β1 treatment were higher in the NRF2-low A549 than those in control cells, and treatment with ROS scavenger blocked TGF-β1-induced cell motility. As an underlying molecular link, NADPH oxidase 4 (NOX4) was associated with higher ROS elevation and cell motility of NRF2-low A549. NOX4 and TGF-β1-inducible NOX4 levels were higher in NRF2-low A549 cells than those in A549. Moreover, the pharmacological inhibition of NOX4 blocked the TGF-β1-induced motility of NRF2-low A549 cells. Collectively, these results indicate that TGF-β1-induced cell motility/migration is facilitated in NRF2-inhibited lung cancer cells and that high levels of NOX4/ROS are associated with enhanced motility/migration.


Lumi-ergometrine--structural identification and occurrence in sclerotia.
The fungus Claviceps purpurea grows on grasses and cereal grains and produces six predominant ergot alkaloids. These toxic substances undergo different transformation reactions during storage and cereal processing. One of these reactions is the addition of water to a double bond in the ergoline skeleton. Since light is required for this process, the substances formed were named lumi-ergot alkaloids. From these, a new asymmetric carbon and consequently two epimers with different polarities are formed. For investigations of lumi-ergot alkaloids, ergometrine was used exemplarily as it represents one of the six ergot alkaloids predominantly formed by Claviceps purpurea. The main reaction product, the less polar compound of the two lumi-ergometrine epimers, was separated by HPLC and unambiguously identified as 10-(S)-lumi-ergometrine using X-ray structural analysis. A HPLC-MS/MS method was developed for the detection of this substance in sclerotia extracts. Using this method, the existence of both epimeric forms of lumi-ergometrine could be proved in the sclerotia. This is the first time that the existence of a lumi-transformation product of ergot alkaloids was proved in naturally grown samples.


Assessment of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification and biotyping of the pathogenic oomycete Pythium insidiosum.
Pythiosis is a life-threatening infectious disease caused by the oomycete Pythium insidiosum. The disease has been increasingly reported worldwide. Most patients with pythiosis undergo surgical removal of an infected organ. Early diagnosis contributes to better prognosis of pythiosis patients. Here, we assessed the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for identification and biotyping of P. insidiosum. A comprehensive set of mass spectra of P. insidiosum was generated to evaluate MALDI-TOF MS for identification and biotyping of P. insidiosum. MALDI-TOF MS accurately identified all 13 P. insidiosum strains tested, at the species level. Mass spectra of P. insidiosum did not match any other microorganisms, including fungi (i.e., Aspergillus species, Fusarium species, and fungal species of the class Zygomycetes), which have similar microscopic morphologies with this oomycete. MALDI-TOF MS- and rDNA sequence-based biotyping methods consistently classified P. insidiosum into three groups: Clade-I (American strains), II (Asian and Australian strains), and III (mostly Thai strains). MALDI-TOF MS has been successfully used for identification and biotyping of P. insidiosum. The obtained mass spectral database allows clinical microbiology laboratories, well-equipped with a MALDI-TOF mass spectrometer, to conveniently identify P. insidiosum, without requiring any pathogen-specific reagents (i.e., antigen, antibody or primers).