Pegylated interferon-α2b and ribavirin decrease claudin-1 and E-cadherin expression in HepG2 and Huh-7.5 cells

Background. Hepatitis C virus (HCV) infection usually results in long-term viremia. Entry of HCV into the he-patocyte requires claudin-1, -6, -9 and occludin. The efficacy of Pegylated interferon-α (PEG-IFN) treatment against HCV infection increased when ribavirin (RBV) was added to the therapeutic scheme. Our aim was to investigate if PEG-IFN plus RBV regulate claudin expression.Material and methods. HepG2, Huh-7 and Huh-7.5 cells were treated with PEG-IFN-α2a or α2b and/or RBV at different times before obtaining the cytosolic, membrane and cytoskeletal fractions. Claudin-1, 3, 4, 6, and 9, E-cadherin and occludin expression was evaluated by Western blot analysis. Transepithelial electrical resistance (TER) was also determined.Results. Claudin-1, 3, 4, 6, E-cadherin and occludin are constitutively expressed mainly in HepG2 cell membrane. Claudin-1 and E-cadherin cell membrane expression diminished after exposure to PEGIFNα2b (50 ng) + RBV(50 μg); the maximal decrease was observed with 200 ng of PEG-IFNα2b + 200 μg of RBV. The effect was less intense with PEG-IFNα2a. The inhibition of claudin-1 and E-cadherin expression in Huh-7 and Huh-7.5 cells was only observed with 200 ng of PEG-IFNα2b + 200 μg of RBV. TER diminished marginally in the HCV containing hepatoma cells with 200 ng of PEG-IFNα2b + 200 ug of RBV. Claudin-1 mRNA expression level was not affected by the combined treatment.Conclusion. The increased therapeutic efficacy of the PEG-IFNα2b plus RBV treatment could be secondary to the inhibition of claudin-1 and E-cadherin cell membrane expression.     

The CHADS2 and CHA2DS2–VASc scores predict new occurrence of atrial fibrillation and ischemic stroke

Background

Early identification of individuals who are at risk of developing atrial fibrillation (AF) and ischemic stroke may enable a closer surveillance and thus prompt initiation of oral anticoagulation for stroke prevention.

Objective

This study sought to investigate whether congestive heart failure, hypertension, age?≥?75 years, diabetes, previous stroke (CHADS₂) and CHA₂DS₂–vascular disease, age 65–74 years, sex category (CHA₂DS₂–VASc) scores can predict new-onset AF and/or ischemic stroke in patients presenting with arrhythmic symptoms.

Methods and results

We prospectively followed up 528 patients (68.5?±?10.6 years, male 46.2 %) presented for assessment of arrhythmic symptoms but without any documented arrhythmia, including AF for development of new-onset AF and/or ischemic stroke. Their mean CHADS₂ and CHA₂DS₂–VASc scores on presentation were 1.3?±?1.3 and 2.3?±?1.5, respectively. After 6.1 years, 89 patients (16.8 %, 2.77 per 100 patient-years) had documented AF, and 65 patients (12.3 %, 2.0 per 100 patient-years) suffered stroke. Both the CHADS₂ (C statistic 0.63, 95 % confidence interval (CI) 0.58–0.67, P?2DS₂–VASc (C statistic 0.63, 95 % CI 0.59–0.67, P?2 (C statistic 0.69, 95 % CI 0.65–0.73, P?2DS₂–VASc (C statistic 0.69, 95 % CI 0.65–0.73, P?Conclusion The CHADS₂ and CHA₂DS₂–VASc scores can be used in patients who presented with arrhythmic symptoms to identify those who are at risk with developing new-onset clinical AF and ischemic stroke for close clinical surveillance and early intervention.     

Cloning and expression and immunogenicity of Helicobacter pylori BabA_2 gene

AIM:To construct a recombinant strain which expresses BabAof Helicobacter pylori(Hpylon)and to study the immunogenicityof BabA.METHODS:BabA_2 DNA was amplified by PCR and insertedinto the prokaryotie expression vector pET-22b( )andexpressed in the BL21(DE3)E.coli strain.Furthermore,BabA immunogenicity was studied by animal test.RESULTS:DNA sequence analysis showed the sequenceof BabA_2 DNA was the same as the one published by GenBank.The BabA recombinant protein accounted for 34.8% of thetotal bacterial protein.The serum from Hpyloriinfectedpatients and Balb/c rniced immunized with BabA itself couldrecognize rBabA.CONCLUSION:BabA recombinant protein may be an potentialvaccine for control and treatment of Hpyloriinfection.     

Expression and significance of Tie-1 and Tie-2 receptors, and angiopoietins-1, 2 and 4 in colorectal adenocarcinoma： Immunohistochemical analysis and correlation with clinicopathological factors

AIM: There is strong evidence that tyrosine kinases are involved in the regulation of tumor progression, cellular growth and differentiation. Recently, many kinds of tyrosine kinase receptors have been reported, among them Tie-1 and Tie-2 receptors constitute a major class. Angiopoietin (Ang)-1 is known as a ligand of Tie-2 tyrosine kinase receptor. The objective of this study was to establish a comprehensive Tie-1 and Tie-2 and Ang-1, 2 and 4 expression profile in human colorectal adenocarcinomas. METHODS: We examined 96 cases of surgically resected human colorectal adenocarcinoma by immunohistochemistry and investigated the statistical correlation between the expressions of Ties and Angs and clinicopathological factors. RESULTS: Among the 96 cases of adenocarcinoma, 87 (90.6%), 92 (95.8%), 83 (86.5%), 89 (92.7%), and 76 cases (79.2%) showed positive staining in the cytoplasm of carcinoma cells for the Tie-1 and Tie-2 and Ang-1, 2 and 4 proteins, respectively. Histologically, the expressions of Ties and Angs were variable. The expressions of Ties and Angs were correlated with several clinicopathological factors, but did not correlate with the presence of lymph node metastasis. Ties and Angs were highly expressed in human colorectal adenocarcinoma cells. CONCLUSION: These findings suggest that the Tie-Ang receptor-ligand complex is one of the factors involved in the cellular differentiation and progression of human colorectal adenocarcinoma.     

NOD2： Ethnic and geographic differences

Investigations into the inheritance of the three risk alleles R702W, G908R and 1007fsInsC in NOD2 associated with susceptibility to Crohn's disease have demonstrated a remarkable amount of heterogeneity across ethnicities and populations, with regional variation across Europe for example, suggesting local founder effects. In non-Caucasian populations Crohn's disease continues to increase in incidence but this increase appears not to be a consequence of variation in NOD2, further advancing the accumulating evidence for other susceptibility loci. Frequencies of the known alleles are compared across populations in health and disease and evidence for additional alleles in NOD2 is reviewed. Based on its position on chromosome 16 coincident with some other autoimmune disease susceptibility localizations, research has targeted NOD2 variation as the potential cause of other autoimmune disorders. While these investigations have mostly returned negative findings, two diseases, Blau Syndrome and Graft versus Host Disease, have been shown to be caused by risk alleles in NOD2. As is frequent in complex disease investigations, some results await validation, but the identification of NOD2 and the differences within and across population raises intriguing questions about the population genetics of the variation at this locus.     

Detection of bcl-2 and bax expression and bcl-2/JH fusion gene in intrahepatic cholangiocarcinoma

AIM:To investigate the relationship between bcl-2 gene and its related protein bax and intrahepatic cholangiocellular carcinoma(CCC). METHODS:Semi-nested in situ PCR(SNISPCR)and imm- unohistochemistry were performed to detect bcl-2/JH fusion gene and bcl-2,bax protein expression in 29 cases of CCC. RESULTS:No bcl-2/JH fusion gene was found in all cases of CCC,72.4% of 29 cases expressed bcl-2 protein.Bcl-2 protein expression was related to histopathological grades (P<0.05).There was no corresponding relationship between bcl-2/JH fusion gene formation and bcl-2 protein expression in CCC(P<0.05).Bax was expressed in 10.3% of 29 cases. The ratio of bcl-2 to bax in normal liver tissues(3.5 to 1)was different from that in tumor tissues(7.0 to 1). CONCLUSION:It is suggested that bcl-2/JH fusion gene formation is not a frequent event and may not play an important role in the pathogenesis of CCC.However,aberrant ratio of bcl-2 to bax protein expression may be involved in the course of tumorigenesis of CCC.Abnormal bcl-2 protein expression may not be solely resulted from bcl-2/JH fusion gene.     

Detection of bcl-2 and bax expression and bcl-2／JH fusion gene in intrahepatic cholangiocarcinoma

AIM: To investigate the relationship between bcl-2 gene and its related protein bax and intrahepatic cholangiocellular carcinoma (CCC). METHODS: Semi-nested in situ PCR (SNISPCR) and immunohistochemistry were performed to detect bcl-2/JH fusion gene and bcl-2, bax protein expression in 29 cases of CCC. RESULTS: No bcl-2/JH fusion gene was found in all cases of CCC, 72.4% of 29 cases expressed bcl-2 protein. Bcl-2 protein expression was related to histopathological grades (P<0.05). There was no corresponding relationship between bcl-2/JH fusion gene formation and bcl-2 protein expression in CCC (P<0.05). Bax was expressed in 10.3% of 29 cases. The ratio of bcl-2 to bax in normal liver tissues (3.5 to 1) was different from that in tumor tissues (7.0 to 1). CONCLUSION: It is suggested that bcl-2/JH fusion gene formation is not a frequent event and may not play an important role in the pathogenesis of CCC. However, aberrant ratio of bcl-2 to bax protein expression may be involved in the course of tumorigenesis of CCC. Abnormal bcl-2 protein expression may not be solely resulted from bcl-2/JH fusion gene.     

T-box 2, a mediator of Bmp-Smad signaling,induced hyaluronan synthase 2 and Tgfβ2 expression and endocardial cushion formation

During early heart development, Tbx2 gene expression is initiated in the cardiac crescent and then becomes restricted to the outflow tract and the atrioventricular region. We identified a Tbx2 regulatory region, enriched in multiple Smad sites, sufficient to reproduce Tbx2 expression patterns overlapping Bmp2 and Bmp4 gene activity in the heart. The role of Tbx2 in cardiogenesis was analyzed by using Cre-LoxP activated Tbx2 transgenic misexpression in chamber myocardium. Ventricular Tbx2 misexpression exhibited an abnormally narrow chamber lumen owing to the expansion of Hyaluronan synthase 2 expression in the ECM or cardiac jelly and the appearance of the endocardial cushions (ECs). Excessive Tbx2 also induced Tgfβ2, which coincided with the outgrowth epithelial-mesenchymal transformed cells in ventricular and atrial tissues modifying cardiomyocyte identity from chamber type to non-chamber type. Tbx2, a central intermediary of Bmp-Smad signaling, has a central part in directing Has2 and Tgfβ2 expression, facilitating EC formation.     

Catalase and PPARγ2 genotype and risk of rheumatoid arthritis in Koreans

Catalase (CAT) and peroxisome proliferator activated receptor-γ2 (PPARγ2) are important regulators of oxidative stress and inflammation, and may contribute to the development of rheumatoid arthritis (RA). We investigated the association between CAT and PPARγ2 genotypes and risk and severity of RA using 474 cases and 400 controls. Genotyping for the −262C→T polymorphism of CAT and the Pro12Ala polymorphism of PPARγ2 was performed by PCR-RFLP analysis. Severity of RA was assessed by the anatomical stage according to Steinbrocker, and a Korean language version of a Health Assessment Questionnaire (KHAQ). No association was observed between CAT and PPARγ2 genotypes and risk of RA. Our results suggest that genetic polymorphisms of CAT and PPARγ2 do not play a significant role in the susceptibility to RA among Koreans.     

Immunopresence and enzymatic activity of nitric oxide synthases, cyclooxygenases and PGE2-9-ketoreductase and in vitro production of PGF2α, PGE2 and testosterone in the testis of adult and prepubertal alpaca (Lama pacos)

This study presents the first evidence for differences in COXs, PGE2-9-ketoreductase and NOSs immunopresence and enzyme activity, and prostaglandin and testosterone production between the testes of adult and prepubertal alpacas. The prepubertal testis immunohistochemical data revealed that COX1 was expressed in spermatogonia and endothelial cells whereas COX2 was present only in the stromal cells. In adult animals, COX2 immunosignals were evidenced in germ cells, as well as both COX1 and -2 in Leydig and Sertoli cells. In adult testes, the spermatogonia, spermatocytes and round spermatids had expression of e- and n-NOS only, whereas elongated spermatids exhibited immunopositivity for i- and e-NOS and Sertoli cells expressed only n-NOS. In prepubertal alpacas, i-NOS was localized in spermatogonia, e-NOS in Sertoli cells and all three NOS isoforms in Leydig cells. PGE2-9-ketoreductase immunopresence was observed in spermatogonia nuclei and cytoplasm of prepubertal testis whereas they were localized in spermatid acrosomal vesicle of adult. The enzymatic data indicated that COX1 activity was higher than COX2 in adult alpaca testis whereas the activity of COX2 was greater than that of COX1 in prepubertal animals. Total NOS and PGE2-9-ketoreductase activities were more extensive in adult alpacas. In vitro hormone production results showed that prepubertal testes released lower amounts of testosterone and PGF2α while PGE2 synthesis was six times more elevated than in in vitro incubated adult testes. Taken together, the data on COX2, i-NOS and PGE2 led us to hypothesize that development in prepubertal male reproductive tissues utilizes a mechanism similar to that of inflammation.     

Expression of MMP—2,TIMP—2 protein and the ratio of MMP—2／TIMP—2 in gallbladder carcinoma and their significance

AIM: To study the correlation between expression of MMP-2, TIMP-2 protein and the ratio of MMP-2/TIMP-2 and clinicalpathological parameters of patients with gallbladder carcinoma.METHODS: Carcinomas (n=45) and polypoid lesions (n=15) of the gallbladder were studied for the expression of MMP-2 and TIMP-2 protein by immunohistochemical avidin-biotin-complex method and image analysis. Clinicalpathological data of patients with gallbladder carcinoma such as histological type, grade of differentiation, level of infiltration, liver invasion and lymph node involvement, etc, were recorded.RESULTS: There was significant difference between the average level (1.123±0.108 VS 1.030±0.054, P=0.002) of MMP-2, the ratio (1.050±0.013 VS0.937±0.078, P=0.003) of MMP-2/TIMP-2 in gallbladder carcinomas and in polypoid lesions of the gallbladder. Significant difference was found between the expression of MMP-2 in early stage and advanced tumors, but there was no correlation between MMP-2 protein expression and histological type, differentiation degree, infiltration level, lymph node involvement or liver invasion. Although no difference was observed between TIMP-2 expression and histological type or differentiation degree, signific ant difference was found between TIMP-2 expression and different Nevin stage, infiltration level, local lymph node involvement or liver invasion (1.168±0.067 VS1.048±0.075, 1.170±0.062 vs 1.039±0.06g, 1.039±0.076 VS1.147±0.083, 1.048±0.074 vs 1.103±0.095, P＜0.05). MMP-2/TIMP-2 ratio did not correlate with histological type, grade of differentiation and liver invasion, but significant differences were found between MMP-2/TIMP-2 ratio and different Nevin stage, infiltration level and lymph node involvement in patients with carcinoma of gallbladder.CONCLUSION: TIMP-2 and MMP-2/TIMP-2 ratio could reflect more accurately biological characteristic of gallbladder carcinoma and MMP-2/TIMP-2 ratio might be a new significant marker in early diagnosis, in the judgment of invasion or metastasis and the estimate of prognosis in patients with gallbladder carcinomas.     

Sodium-glucose co-transporter 2 inhibitors and heart failure—the present and the future

Heart Failure Reviews - Type 2 diabetes mellitus is associated with an increased risk of heart failure. The prevalence of type 2 diabetes mellitus is on an upward trend. Heart failure represents...     

Do Dogs and Cats Passively Carry SARS-CoV-2 on Hair and Pads?

The epidemiological role of domestic animals in the spread and transmission of SARS-CoV-2 to humans has been investigated in recent reports, but some aspects need to be further clarified. To date, only in rare cases have dogs and cats living with COVID-19 patients been found to harbour SARS-CoV-2, with no evidence of pet-to-human transmission. The aim of the present study was to verify whether dogs and cats act as passive mechanical carriers of SARS-CoV-2 when they live in close contact with COVID-19 patients. Cutaneous and interdigital swabs collected from 48 dogs and 15 cats owned by COVID-19 patients were tested for SARS-CoV-2 by qRT-PCR. The time elapsed between owner swab positivity and sample collection from pets ranged from 1 to 72 days, with a median time of 23 days for dogs and 39 days for cats. All samples tested negative, suggesting that pets do not passively carry SARS-CoV-2 on their hair and pads, and thus they likely do not play an important role in the virus transmission to humans. This data may contribute to confirming that the direct contact with the hair and pads of pets does not represent a route for the transmission of SARS-CoV-2.     

Immunoglobulin and β2-microglobulin concentrations in bronchoalveolar lavage of children and adults

Immunoglobulins play an important role in the pulmonary host defense, but little information is available about immunoglobulin and 2-microglobulin concentrations in the lung of normal children. Using bronchoalveolar lavage (BAL) we have studied immunoglobulin and 2-microglobulin levels in 30 children 3–15 years old undergoing elective surgery for nonpulmonary illnesses and in 15 healthy adult volunteers. BAL was performed with 3 × 1 ml/kg of body weight normal saline through an endotracheal tube after induction of anesthesia in children and under local anesthesia in adults. Similar concentrations of IgA and IgG were found in BAL fluid of children and adults even though serum levels were lower in children. As comparable results were obtained for albumin, a serum-derived protein, these data suggest that the permeability of the alveolar membrane is higher in children. IgE and IgM were detected in BAL fluid in only a fraction of children. 2-microglobulin levels were higher in both blood and BAL fluid of children. These data provide the first reference data for immunoglobulin and 2-microglobulin in children and can serve as a basis for future studies of children with pulmonary diseases.     

Laparoscopic liver sectionectomy 2 and 3 (LLS 2 and 3): towards the “gold standard”

Background

The purpose of this article is to define the state of the art in laparoscopic liver sectionectomy 2 and 3 (LLS 2 and 3) in order to advance the good option towards the “gold standard”.

Methods

Based on a large review of the literature as well as on our personal experience the authors define clearly: the feasibility and the effectiveness of LLS 2 and 3.

Results

In this review the conversion rate was <4%, the histological positive margins was <0.8%, and the mortality was inferior to 0.8%.

Conclusion

The LLS 2 and 3 seem equivalent or perhaps better option compared with the same intervention performed by laparotomy and can be proposed as primary with a grade C recommendation.     

Origins of R2? and white matter

Estimates of the apparent transverse relaxation rate () can be used to quantify important properties of biological tissue. Surprisingly, the mechanism of dependence on tissue orientation is not well understood. The primary goal of this paper was to characterize orientation dependence of in gray and white matter and relate it to independent measurements of two other susceptibility based parameters: the local Larmor frequency shift (f_L) and quantitative volume magnetic susceptibility (Δχ). Through this comparative analysis we calculated scaling relations quantifying (reversible contribution to the transverse relaxation rate from local field inhomogeneities) in a voxel given measurements of the local Larmor frequency shift. is a measure of both perturber geometry and density and is related to tissue microstructure. Additionally, two methods (the Generalized Lorentzian model and iterative dipole inversion) for calculating Δχ were compared in gray and white matter. The value of Δχ derived from fitting the Generalized Lorentzian model was then connected to the observed orientation dependence using image-registered optical density measurements from histochemical staining. Our results demonstrate that the and f_L of white and cortical gray matter are well described by a sinusoidal dependence on the orientation of the tissue and a linear dependence on the volume fraction of myelin in the tissue. In deep brain gray matter structures, where there is no obvious symmetry axis, and f_L have no orientation dependence but retain a linear dependence on tissue iron concentration and hence Δχ.In many neurological diseases such as multiple sclerosis, Alzheimer’s, and Parkinson, and in conditions following traumatic brain injury, microstructural changes occur in gray and white matter (1–4). One method for quantifying these microstructural changes is the mapping of the effective transverse relaxation rate (). Along with the longitudinal relaxation rate (R₁) and transverse relaxation rate (R₂), has been viewed as a fundamental MRI tissue parameter, affected by several factors including myelin content (5, 6), endogenous ferritin-based (Fe³⁺) iron (7, 8), tissue microstructure (6), and paramagnetic, blood deoxyhemoglobin (9). However, a number of recent studies have reported a somewhat surprising dependence of on tissue orientation, at least in white matter (10–12). The purpose of this paper was to investigate the mechanisms that could contribute to this orientation dependence of in both gray and white matter. Because is influenced by magnetic field perturbations, we examined the role of local Larmor frequency shift (f_L) and quantitative magnetic susceptibility (Δχ), parameters that relate field and frequency. Through this analysis we identified unique scaling relations that relate to the local Larmor frequency shift calculated after removal of macroscopic field inhomogeneities. Additionally, we compared two methods for computing Δχ in gray and white matter: (i) fitting the Generalized Lorentzian (GL) model of field perturbers (13) to f_L measured at multiple brain orientations and (ii) magnitude-regularized dipole inversion (14). The difference between these two estimates represents the local frequency shift due to a cylindrical, axon geometry and is a marker of axonal integrity.Having modeled the orientation dependence, we next examined the effect of myelin and iron on both and Δχ. We demonstrated a linear correlation between these quantities and optical density (OD) derived from diaminobenzidine (DAB)-enhanced Perls stain (sensitive to ferritin-based iron) for cortical gray matter. Similarly, using OD derived from solochrome cyanine-R (ScR)–stained slides in rat brain major white matter fiber regions, we showed strong positive correlations between the transverse relaxation constants, R₂ and , and myelin density. Taken together, our results demonstrate that observed values in ex vivo brains can be explained by a sinusoidal dependence on tissue microstructure orientation in conjunction with a linear dependence on the myelin concentration in cortical gray and white matter. In deeper gray matter structures with no preferred symmetry axis, does not have an orientation dependence but retains a linear dependence on iron concentration.