Engineering human ACE2 to optimize binding to the spike protein of SARS coronavirus 2

The spike(S)protein of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)binds angiotensin-converting enzyme 2(ACE2)on host cells to initiate entry,and soluble ACE2 is a therapeutic candidate that neutralizes infection by acting as a decoy.By using deep mutagenesis,mutations in ACE2 that increase S binding are found across the interaction surface,in the asparagine 90-glycosylation motif and at buried sites.The mutational landscape provides a blueprint for understanding the specificity of the interaction between ACE2 and S and for engineering high-affinity decoy receptors.     

Transfer of the amyloid β and/or of β-amyloid precursor protein of the fetus with trisomy 21 to the maternal blood stream and its possible contribution to the pathogenesis of the maternal Alzheimer's Disease

Down Syndrome (DS) is the most frequent genetic pathology. It affects 1 out of every 800 newborn babies. Approximately between a 90% and a 95% of all the cases of DS are attributed to a trisomy in chromosome 21. One of the genes contained in this chromosome is the gene of β-amyloid precursor protein (βAPP). The metabolism of this protein yields, among others, the amyloid beta peptides made up of 40 amino acids (Aβ40) and 42 amino acids (Aβ42). The evidence that is derived from several sources - genetic, among them - suggests that the Aβ participates in the pathogenesis of Alzheimer’s Disease (AD).It is worth pointing at the fact that the transfer of cells, extracellular chromosomal material and some proteins from the fetus to the mother and vice versa has been widely described. The transfer rate from the fetus to the mother is higher when the mother is carrying a baby with trisomy 21. This has led to the hypothesis that sets forth that during the gestation of a baby with DS there is a greater fetomaternal transfer of cells and of products of the genes of chromosome 21 - among them, βAPP and its metabolites Aβ40 and Aβ42. It is possible to speculate on the possible contribution of the fetal components - among them, Aβ - to the higher risk of suffering AD, which has been reported in a subpopulation of women who have given birth to children with DS. On the other hand, the detection of the βAPP - mainly intracellular - and of the β amyloid peptides in maternal blood and urine during the early stages of gestation could be taken as a potential non invasive biochemical prenatal marker of DS.     

Suggestive evidence that Fc variants of IgG2 and FcγRIIa loci interact to contribute to the risk of prostate cancer

GM and KM allotypes—hereditary antigenic variants of immunoglobulin γ and κ chains, respectively—and the genetic variants of activating Fcγ receptors (FcγR) are associated with the immunobiology of several malignant diseases, but their role in susceptibility to prostate cancer has not been examined. This investigation aimed to determine whether these genes—individually or in particular epistatic combinations—contribute to the risk of prostate cancer. We genotyped DNA from 200 Caucasian patients with prostate cancer and 185 healthy controls (matched for age, race, gender, and geography) for several GM, KM, FcγRIIa, and FcγRIIIa alleles by molecular methods. None of the genotypes by itself was associated with the risk of prostate cancer. However, particular alleles at GM 23 and FcγRIIa loci interactively contributed to the risk of this malignancy (p = 0.031), the odds ratios ranging from 0.44 in subjects homozygous for the GM 23− allele at the IgG2 locus and for the histidine allele at the FcγRIIa locus to 2.86 in subjects homozygous for the GM 23+ allele at the IgG2 locus and the histidine allele at the FcγRIIa locus. To our knowledge, this is the first report implicating GM and FcγR loci as risk/protective factors for prostate cancer. Additional, independent, studies are warranted to confirm and extend these findings.     

Study of DNA damage induction and repair capacity of fresh and cryopreserved lymphocytes exposed to H2O2 and γ-irradiation with the alkaline comet assay

The alkaline SCGE assay was evaluated for use with cryopreserved lymphocytes in order to obtain results similar to the freshly isolated ones. The induction of DNA damage as well as the repair capacity of γ-rays and H₂O₂ exposed cryopreserved human lymphocytes was found to be the same to that of the freshly isolated. Human lymphocytes (fresh or cryopreserved) responded differently to the effects of γ-irradiation if compared to the H₂O₂ treatment. The distribution of DNA damage among γ-irradiated lymphocytes was more homogeneous compared to H₂O₂, both in freshly isolated and in cryopreserved cells. 2.4 μg/ml phytohemagglutinin at the start of a 2-h incubation in RPMI of cryopreserved samples gave similar DNA repair and distribution patterns to the 2-h post-exposure incubation of freshly isolated lymphocytes. H₂O₂-induced DNA damage was not repaired completely. However, the repair of γ-rays-induced DNA damage was more efficient. These findings confirm the different mode of action of the two agents on the induction of DNA damage, as well as, the different response of the lymphocytes' DNA repair system.     

Does the oxidation of methionine in thrombomodulin contribute to the hypercoaguable state of smokers and diabetics?

The leading cause of premature death in smokers is cardiovascular disease. Diabetics also suffer from increased cardiovascular disease. This results, in part, from the hypercoagulable state associated with these conditions. However, the molecular cause(s) of the elevated risk of cardiovascular disease and the prothrombotic state of smokers and diabetics remain unknown. It is well known that oxidative stress is increased in both conditions. In smokers, it is established that oxidation of methionine residues takes place in alpha(1)-antitrypsin in lungs and that this leads to emphysema. Thrombomodulin is a key regulator of blood clotting and is found on the endothelium. Oxidation of methionine 388 in thrombomodulin is known to slow the rate at which the thrombomodulin-thrombin complex activates protein C, a protein which, in turn, degrades the factors which activate thrombin and lead to clot formation. In analogy to the cause of emphysema, it is hypothesized that oxidation of this methionine is elevated in smokers relative to non-smokers and, perhaps, in conditions such as diabetes that impose oxidative stress on the body. Evidence for the hypothesis that such an oxidation and concomitant reduction in activated protein C levels would lead to elevated cardiovascular risk is presented.     

Place of genotyping in addition to the phenotype and the assay of serum α-1 antitrypsin

The diagnosis of deficiency of alpha-1 antitrypsin (A1AT) is based on isoelectric focusing of serum proteins and the extent of serum. However, the focusing is technically difficult and a greatly reduced concentration in abnormal A1AT tapeless does not differentiate an unstable variant of a variant called 'null' (that is to say without any phenotypic expression) to 'heterozygous' state. In this study, we compared the results of the assay, the phenotype and genotype of A1AT in 50 patients. Normal A1AT alleles (Pi*M1 to Pi*M4) or loss of the most common (Pi*S and Pi*Z) were clearly identified in phenotyping. However, genotyping was necessary to characterize: (i) certain alleles rarer A1AT (S-Munich, X-Christchurch); (ii) a null allele and; (iii) two new alleles A1AT not yet described in the literature. In conclusion, although the A1AT genotyping is generally not necessary, it is necessary to resolve complex cases and to obtain witnesses validated for isoelectric focusing.     

Lectin binding patterns of alveolar epithelium and subepithelial seromucous glands of the bronchi in sepsis and controls – an approach to characterize the non-specific immunological response of the human lung to sepsis

A delayed or deficient immunological protection as well as an overstimulation of the mucosal immune system may act as possible additional promoters of sepsis-induced lung injury in patients suffering from a severe septic condition. Lectin-binding patterns in pulmonary tissue samples obtained at autopsy from septic patients and control individuals were studied using 11 carbohydrate-specific lectins (Con A, UEA, GSA I, GSA II, MPA, PNA, Jac, WGA, MAA, LPA, and SNA). There were no differences in the secretory product of serous parts of bronchial glands detectable in the two study groups, whereas lectin binding patterns of alveolar epithelium and mucous parts of subepithelial seromucous glands were different in sepsis cases when compared to controls. Apart from differences in binding sites for alpha-mannose, N-acetyl-neuraminic acid and alpha-(2-6)-galactose (as detected by different expression for Con A, MAA and SNA) in the two study groups, the main finding was that no binding sites for alpha-N-acetyl-galactosamine (as investigated by MPA immunoreactivity) could be detected on alveolar epithelial cells and mucous parts of subepithelial seromucous glands in sepsis cases in contrast to the presence of such binding sites in the control cases. We hypothesize that the finding of an altered secretory product of alveolar epithelial cells and bronchial glands in sepsis may be a result of specific carbohydrate deprivation or consumption, respectively, possibly due to direct bacterial effects or pathogenetic events in response to bacterial toxins during the complex cascade of the host's systemic inflammatory response in sepsis. The altered type of mucus glycoprotein physiologically secreted by alveolar epithelium and mucous parts of subepithelial seromucous glands of the bronchi with subsequent loss of a considerable proportion of protection of the mucosal barrier in sepsis may play an important additional role in the development of sepsis-induced lung injury.     

Localization of the gene encoding the α2 subunit of the human VLA-2 receptor to chromosome 5q23-31

The ₂ subunit of the VLA-2 receptor (CD49B) was mapped to human chromosome 5 by several independent approaches. First, the expression of the ₂ subunit at the protein level was investigated in a panel of human-mouse hybrid cell lines. Cell surface expression was detected by indirect immunofluorescence with monoclonal anti-₂ antibody 12F1. Intracellular ₂ antigen was detected by immunostaining of whole cell extracts or of immunoprecipitated 12F1 antigen with the monoclonal antibodies 3H8 and 5C5. Second, the presence of human genomic ₂ sequences in the panel of human-mouse hybrids was detected by PCR, using primers derived from the published ₂ cDNA sequence. The specificity of the amplification product was shown by direct sequencing. The results of the PCR study were confirmed by amplifying aCD14 gene fragment, known to map to chromosome 5. Finally, in situ hybridization with a³H-labeled 1040-bp cDNA probe, also obtained by PCR, confirmed and refined the localization ofCD49B on chromosome 5 at q23-31.     

Relevances of microfilament and microtubule to the adhesion properties of the HCC onto the Collagen IV/ Laminin coated surface

INTRODUCTION　　Tumorinvasionisclearlyanactiveprocessundergoingthreestages:attach-ment,migrationandmetastasis〔1〕.Infacts,tumorinvasioncouldbeconsideredady-namicadhesiveprocess,whichcontainedmuchcomplexandsubtlephysiologicalandbiochemicalinteractionsbetw…     

Interferons and scleroderma-a new clue to understanding the pathogenesis of scleroderma?

Scleroderma or systemic sclerosis (SSc) is a complex disease characterized by vasculopathy and deregulated immune and fibroblast activation. The resulting excessive production of collagens and other extracellular matrix proteins by fibroblasts as well as the inflammatory response leads to the development of scleroderma. Recently, some emerging data have been showing a possible link between the type I and II interferons (IFNs) and SSc pathogenesis. IFNs are well-known immunomodulators and inhibitors of collagen production. However, IFN therapy also has been implicated in the development or exacerbation of several autoimmune diseases, including SSc. Some studies also showed an increase mRNA and protein levels of IFNs and several interferon stimulated genes in cells and tissues from SSc patients. In this review we discuss about a possible role for IFNs in SSc development and pathogenesis.     

The Analysis to the Influence of the Mass Athletic Dance on the Middle and old aged Health

The article gives the analysis to the mass athletic dances and its influence on the health of the middle and old-aged people. The research shows, through contrasting, observing and physiologically analyzing, that there is an improvement in lowering people‘s blood viscosity and increasing fats catabolism, raising the serum level as well as lowering the serum density, if these people take part in the mass athletic dances regularly for a long time.     

Effects of the administration of pentoxifylline and prednisolone on the evolution of portal fibrogenesis secondary to biliary obstruction in growing animals: immunohistochemical analysis of the expression of TGF- β and VEGF

OBJECTIVE:

During the neonatal and infancy periods, some chronic liver diseases may lead to progressive hepatic fibrosis, which is a condition that can ultimately result in the loss of organ function and severe portal hypertension necessitating hepatic transplantation. In a previous report, pharmacological interventions were demonstrated to modulate hepatic fibrosis induced by bile duct ligation in young rats. The administration of pentoxifylline or prednisolone, or the combination of both, resulted in reduced fibrogenesis in portal spaces. The objectives of the present study were to evaluate the expression of transforming growth factor β and vascular endothelial growth factor after bile duct ligation in young rats and to assess the effect of those same drugs on cytokine expression.

METHODS:

In this experimental study, 80 young rats (21 or 22 days old) were submitted either to laparotomy and common bile duct ligation or to sham surgery. The animals were allocated into four groups according to surgical procedure, and the following treatments were administered: (1) common bile duct ligation + distilled water, (2) sham surgery + distilled water, (3) common bile duct ligation + pentoxifylline, or (4) common bile duct ligation + prednisolone. After 30 days, a hepatic fragment was collected from each animal for immunohistochemical analysis using monoclonal antibodies against transforming growth factor β and vascular endothelial growth factor. Digital morphometric and statistical analyses were performed.

RESULTS:

The administration of pentoxifylline reduced the transforming growth factor β-marked area and the amount of transforming growth factor β expressed in liver tissue. This effect was not observed after the administration of prednisolone. There was a significant reduction in vascular endothelial growth factor expression after the administration of either drug compared with the non-treatment group.

CONCLUSIONS:

The administration of pentoxifylline to cholestatic young rats resulted in the diminished expression of transforming growth factor β and vascular endothelial growth factor in liver tissue. The administration of steroids resulted in the diminished expression of vascular endothelial growth factor only. These pathways may be involved in hepatic fibrogenesis in young rats submitted to bile duct ligation and exposed to pentoxifylline or prednisolone.     

Contributions of the Kölliker–Fuse nucleus to coordination of breathing and swallowing

Herein we compare the effects of perturbations in the Kölliker–Fuse nucleus (KFN) and the lateral (LPBN) and medial (MPBN) parabrachial nuclei on the coordination of breathing and swallowing. Cannula was chronically implanted in goats through which ibotenic acid (IA) was injected while awake. Swallows in late expiration (E) always reset while swallows in early inspiration (I) never reset the respiratory rhythm. Before cannula implantation, all other E and I swallows did not reset the respiratory rhythm, and had small effects on E and I duration and tidal volume (V_T). However, after cannula implantation in the MPBN and KFN, E and I swallows reset the respiratory rhythm and increased the effects on I and E duration and V_T. Subsequent injection of IA into the KFN eliminated the respiratory phase resetting of swallows but exacerbated the effects on I and E duration and V_T. We conclude that the KFN and to a lesser extent the MPBN contribute to coordination of breathing and swallowing.     

Contributions of vision–proprioception interactions to the estimation of time-varying hand and target locations

We investigated the relative importance of vision and proprioception in estimating target and hand locations in a dynamic environment. Subjects performed a position estimation task in which a target moved horizontally on a screen at a constant velocity and then disappeared. They were asked to estimate the position of the invisible target under two conditions: passively observing and manually tracking. The tracking trials included three visual conditions with a cursor representing the hand position: always visible, disappearing simultaneously with target disappearance, and always invisible. The target’s invisible displacement was systematically underestimated during passive observation. In active conditions, tracking with the visible cursor significantly decreased the extent of underestimation. Tracking of the invisible target became much more accurate under this condition and was not affected by cursor disappearance. In a second experiment, subjects were asked to judge the position of their unseen hand instead of the target during tracking movements. Invisible hand displacements were also underestimated when compared with the actual displacement. Continuous or brief presentation of the cursor reduced the extent of underestimation. These results suggest that vision–proprioception interactions are critical for representing exact target–hand spatial relationships, and that such sensorimotor representation of hand kinematics serves a cognitive function in predicting target position. We propose a hypothesis that the central nervous system can utilize information derived from proprioception and/or efference copy for sensorimotor prediction of dynamic target and hand positions, but that effective use of this information for conscious estimation requires that it be presented in a form that corresponds to that used for the estimations.     

Is it possible to predict the long-term response to venlafaxine with the use of biological markers and psychophysiological methods?

INTRODUCTION: The present study investigated whether it is possible to predict the medium term response to venlafaxine using biological markers and psychophysiological methods. MATERIAL: Fourteen (14) patients aged 21-60 years suffering from Major Depression according to DSM-IV were included in the study. METHODS: The SCAN v 2.0 and the IPDE were used to assist clinical diagnosis. Patients were investigated with electrooculogram (EOG), Pattern-Reversal Visual Evoked Potentials (PR-VEPs), Dexamethasone Suppression Test (DST), D-fenfluramine Challenge Test, and brain Single Photon Emission Tomography (SPECT). Venlafaxine 150-225 mg per os daily was administered. The follow-up period was 2 years. STATISTICAL ANALYSIS: Chi-square test and ANOVA were used for the analysis of data. RESULTS: There was a lower left globus pallidus regional cerebral blood flow in patients with better response. On the contrary, chronic patients were closer to normality. DISCUSSION: The results of the current study provide preliminary evidence concerning our ability to predict response to venlafaxine and to understand its way of action.