The roles of Transforming growth Factor Type β<Subscript>3</Subscript>(TGF-β<Subscript>3</Subscript>) and mast cells in the pathogenesis of scleroderma

Scleroderma is a connective tissue disorder characterised by excessive accumulation of collagen in the skin and internal organs. The most likely explanation for this process is local activation of collagen synthesis from fibroblasts. Our intention was to elucidate whether TGF-₃ and mast cells play a pathogenic role in abnormal connective tissue formation in scleroderma. In this study, skin biopsies from 20 patients with scleroderma and five from healthy individuals were studied by an indirect immunoperoxidase technique to determine the immunoreactivity of TGF-₃ in the dermis. In addition, skin samples were stained with toluidine blue to count the number of mast cells in scleroderma, and tissues were examined under the electron microscope to evaluate the ultrastructural changes. Increased TGF-₃ immunoreactivities were detected in the dermis in the patient's skin, suggesting the presence of a subpopulation responsible for the increased collagen production. Mast cell counts in the skin of patients with scleroderma were significantly greater (19.2 ± 4.1/unit) than those of normal controls (4.4 ± 1.2/unit). Ultrastructural observations indicated that there is a close relationship between the mast cells and fibroblasts. These results suggest that fibrosis in scleroderma could evolve through the activation of fibroblasts and the regulatory mechanisms that appear to modulate the behavior of these cells with respect to collagen production.AcknowledgementWe are grateful for the support of Professor M. Kaya in our work with the electron microscope and thank Professor E. Erten for supplying tissue samples.     

Prevalence of Anticardiolipin and Anti-β<Subscript>2</Subscript>-Glycoprotein I Antibodies in Celiac Disease

The aim of this study was to evaluate the prevalence of anticardiolipin antibodies (aCL) and anti-β₂-glycoprotein I antibodies (aβ2GPI) in patients with celiac disease and to analyze the clinical features of antiphospholipid syndrome in these patients. We conducted a prospective case-control study based on the evaluation of IgG, IgM and IgA aCL, and IgG and IgA aβ2GPI in celiac disease patients and in controls. All patients were asked about any occurrence of thrombotic manifestations. In addition, women were asked about pregnancy morbidity. Fifty celiac disease patients and 50 healthy controls were studied. IgM aCL were not detected in study group or in controls. IgG aCL were found in two patients and in one control. IgA aCL were significantly more frequent in celiac disease patients compared with controls (13/50 (26%) vs. 2/50 (4%), p=0.004, OR [95% CI]=9.09 [1.81–50]). There was no statistically significant difference for the prevalence of IgG and IgA aβ2GPI between patients and controls. Clinical features of antiphospholipid syndrome were noted in two patients with negative antibodies. Prevalence of IgM and IgG aCL and of aβ2GPI were not increased in celiac disease. IgA aCL were more frequently detected in celiac disease. However, no clinical features of antiphospholipid syndrome were noted.     

Pharmacological Stimulation of β <Subscript>2</Subscript>-adrenergic Receptors (β <Subscript>2</Subscript>AR) Enhances Therapeutic Effectiveness of β <Subscript>1</Subscript>AR Blockade in Rodent Dilated Ischemic Cardiomyopathy

Background. We have reported that β ₂ adrenoreceptor (β ₂AR) stimulation is anti-apoptotic, and has strong beneficial effect on cardiac remodeling in an experimental model of post myocardial infarction chronic heart failure (CHF) in rats. Here we investigate whether the addition of chronic pharmacological β ₂AR stimulation enhances the therapeutic effects of β ₁AR blockade on cardiac remodeling in the same model. Methods and Results. Metoprolol, a β ₁AR blocker, given alone (β ₁) or in combination with β ₂AR agonist, fenoterol (β ₁β ₂) were administered to rats via drinking water for 6 weeks, beginning 2 weeks following permanent coronary ligation. Progressive left ventricular (LV) remodeling of untreated animals, assessed by repeated echocardiography, occurred during the observation time, i.e., 42% and 25% increases in end-systolic and end-diastolic LV volumes respectively, 27% fall in ejection fraction, and 35% infarct expansion. Pressure-volume loop analyses at 2d and 8th post infarction weeks showed continuous deterioration of systolic and diastolic functions and arterio-ventricular mismatch. Histological evaluation at the end of 8 weeks revealed the MI expansion and hypertrophy of cardiomyocytes. β ₁β ₂ prevented LV remodeling, MI expansion and cardiomyocytes hypertrophy to a greater degree than β ₁, due, in large part, to a vasodilatory effect of β ₂AR stimulation and thus improvement of arterio-ventricular mismatch. The abnormal diastolic performance improved only in β ₁β ₂. β ₁β ₂ treatment reduced myocardial apoptosis throughout myocardium, but β ₁ reduced apoptopsis only in the areas remote from MI. Conclusion. The therapeutic effects of chronic β ₁AR blockade on cardiac remodeling of heart failure are enhanced and extended when supplemented with β ₂AR stimulation. This research was supported by the Intramural Research Program of the National Institute on Aging, NIH.     

Effects of the inhibition of cytosolic phospholipase A<Subscript>2</Subscript>α in non-small cell lung cancer cells

Purpose  

The aim of this study was to investigate the expression of cPLA₂α in non-small lung cancer cell lines and tissues, and we sought to determine the in vitro effects of the pyrrolidine-2 inhibitor on cPLA₂α sensitivity in three different non-small lung cancer cell lines.     

Shear stress inhibition of H<Subscript>2</Subscript>O<Subscript>2</Subscript> induced p66<Superscript>Shc</Superscript> phosphorylation by ASK1–JNK inactivation in endothelium

Shear stress protects endothelium from a variety of risk factors for vascular disease. Here, we demonstrate a novel mechanism whereby shear stress inhibited reactive oxygen species (ROS)-triggered signaling cascades in endothelial cells. Stimulation of bovine aortic endothelial cells (BAECs) with H₂O₂ induced a 3.07-fold increase in p66^Shc phosphorylation. This response was fully blocked by pretreatment of cells with specific JNK but not p38 or ERK MAP kinase inhibitor. Further study showed that knocking down of apoptosis signal-regulating kinase 1 (ASK1) by siRNA transfection in cells dramatically inhibited phosphorylation of JNK and p66^Shc elicited by H₂O₂. Pre-perfusion of BAECs cultured in silastic tubes with laminar flow generated by a servo-pump system for 30 min also significantly suppressed H₂O₂-induced phosphorylation of p66^Shc. This was accompanied by quantitatively similar inhibition of ASK1 and JNK phosphorylation and activation. These results suggested that shear stress protects endothelium against oxidant stress by suppression of ASK1–JNK-mediated p66^Shc phosphorylation.     

Anti-β<Subscript>2</Subscript>-glycoprotein I in Sjogren’s syndrome is associated with parkinsonism

The nervous system may be involved in up to 30% of patients with Sjogren’s syndrome (SS). We describe three patients with Sjogren’s syndrome and a concomitant parkinsonian syndrome. Elevated titers of anti-β₂-glycoprotein I IgG were found in the serum of all three patients. This autoantibody is strongly associated with anticardiolipin (aCL) antibodies, antiphospholipid syndrome (APS), and thromboembolic phenomena, but its role in the pathogenesis of the parkinsonian disorder in SS is unclear. These patients may present a subtype of SS patients in which the presence of aCL antibodies is associated with central nervous system involvement predominantly in the basal ganglia. Sharon Hassin-Baer and Levy Yair contributed equally to this work.     

Clinical Efficacy of Inhaler Devices Containing β<Subscript>2</Subscript>-Agonist Bronchodilators in the Treatment of Asthma

Background: A number of different inhaler devices are available to deliver β₂-adrenoceptor agonist (β₂-agonist) bronchodilators in asthma. These include hydrofluoroalkane or chlorofluorocarbon (CFC)-free propelled pressurized metered-dose inhalers (pMDIs), many dry powder inhalers and breath-actuated inhalers. Objective: To determine the clinical efficacy of all available hand-held inhaler devices compared with the standard CFC-containing pMDI for the delivery of short-acting β₂-agonist bronchodilators in nonacute asthma in both children and adults. Methodology: A systematic review and meta-analysis was carried out of all available randomized, controlled trials (RCTs) using the standard pMDI compared with any other hand-held inhaler device, delivering short-acting β₂-agonist bronchodilators in patients with stable asthma. Results: One hundred and eighteen RCTs were included in this review. No clinical differences were found between the standard CFC-containing pMDI and 12 other hand-held inhaler devices for most outcome measures. We found no evidence of clinical differences between studies using either a 1 : 1 (pMDI: another inhaler) or a 2 : 1 dosing ratio. Conclusions: In patients with stable asthma, short-acting β₂-agonist bronchodilators in standard CFC-pMDIs are as effective as any other hand-held inhaler device; therefore the cheapest available device that the patient is able to use should always be considered. Pharmaceutical companies should in future submit to regulatory authorities clinical outcome data (as opposed to in vitro data) in support of any dosing schedules greater than 1 : 1 when compared with the standard pMDI. Clinical effectiveness studies that use an intention-to-treat analysis and report more patient-centered outcomes are required.     

rAAV2-TGF-β<Subscript>3</Subscript> Decreases Collagen Synthesis and Deposition in the Liver of Experimental Hepatic Fibrosis Rat

Background/Aims  

Hepatic fibrosis is one kind of common wound-healing response to chronic liver injury. Transforming growth factor (TGF)-β₃ performs an anti-fibrosis function under certain conditions such as pancreatic fibrosis and wound healing. This study aimed at investigating the effect of TGF-β₃ on the histology in the liver of rat with liver fibrosis.     

An Increase of Blood Anti-β<Subscript><Emphasis Type="Bold">2</Emphasis></Subscript>-glycoprotein-I Antibody Associates with Cerebral Hemorrhage in Patients without Antiphospholipid Antibody Syndrome

Although an increase of blood anti-beta2-glycoprotein I antibody (aGPI) is known to raise the risk for cerebral ischemia in patients without antiphospholipid antibody syndrome or systemic lupus erythrematosus, there is no case of cerebral hemorrhage mentioned yet. We reported 6 patients who suffered from nontraumatic cerebral hemorrhage who were identified to have an increase of aGPI. The frequency of increased aGPI in cerebral hemorrhage was 9.7%. Hematoma was predilected at putamen in patients with increased aGPI. The activity of coagulation factors was normal. Multiple brain infarcts compatible with small artery occlusion were illustrated in all of them. Accordingly, cerebral hemorrhage may arise out of small artery vasculopathy in these patients. These results highlight a relationship between autoimmunity, such as aGPI, and cerebral hemorrhage, especially in the case of an absence of traditional risk factor.     

Incidence of ischemic stroke and systemic embolism in patients with hypertrophic cardiomyopathy,nonvalvular atrial fibrillation,CHA<Subscript>2</Subscript>DS<Subscript>2</Subscript>-VASc score of ≤1 and without anticoagulant therapy

Data on the risk of ischemic stroke and systemic embolism (iSSE) events in patients with nonvalvular atrial fibrillation (NVAF), a CHA₂DS₂-VASc score of ≤1, hypertrophic cardiomyopathy (HCM), and without anticoagulant therapy are still lacking. The aim of this study was to investigate the incidence of iSSE events in these patients. We consecutively screened medical records of patients with HCM and NVAF referred to Fuwai Hospital between January 1994 and March 2014. The primary end point was iSSE events, defined as a composite of ischemic stroke and systemic embolism. Follow-up was carried out to ascertain end point status. Medical records of 522 patients with NVAF and HCM were screened. A total of 108 patients (20.7 %) with a CHA₂DS₂-VASc score of ≤1 and without anticoagulant therapy were enrolled and constituted our study population. After a median follow-up of 2.4 years (range 0.6–14.1 years; 376.2 patient-years), ischemic stroke occurred in 2 patients, resulting in death of 1 patient in the first year and paralysis of the other patient in the fourth year. No other iSSE events occurred. The incidence of iSSE was 0.9 % [95 % confidence interval (CI) 0.0–5.0 %] in the first year, and 0.5 % per 100 patient-years (95 % CI 0.1–1.9 %). The risk of iSSE events seems low in patients with NVAF, a CHA₂DS₂-VASc score of ≤1, HCM, and without anticoagulant therapy. Multicenter studies with sizeable study populations are needed to validate the risk of iSSE events in these patients.     

Donepezil improves the cognitive impairment in a tree shrew model of Alzheimer’s disease induced by amyloid-β<Subscript>1–40</Subscript> via activating the BDNF/TrkB signal pathway

Alzheimer’s disease (AD) is a chronic neurodegenerative disorder which can contribute to memory loss and cognitive damage in the elderly; moreover, evidence from clinical and animal studies demonstrated that AD always exhibit severe cognitive deficits. However, the effects of donepezil medications on cognition are controversial. Additionally, it is unclear whether donepezil can protect neurons to improve cognitive function through the brain-derived neurotropic factor (BDNF)/tyrosine receptor kinase B (TrkB) signalling pathway in the tree shrew (TS), which has a closer evolutionary relationship to primates than rodents. Here, we designed a study on an amyloid-β_1–40 (Aβ_1–40)-induced TS model of AD and investigated the molecular mechanism by which donepezil protects neurons and improves cognitive function through activating the BDNF/TrkB signalling pathway. The results showed that donepezil could rescue Aβ_1–40-induced spatial cognition deficits, and reverse Aβ_1–40-induced temporal horn along with ADC enlargement in the TS brain. Meanwhile, it suppressed Aβ_1–40-induced neuronal damage and loss of body weight. Intriguingly, donepezil could increase the choline acetyl transferase (ChAT) expression level and reduce the fibrillary acid protein (GFAP) expression level in the hippocampus and cortex of TS. Additionally, donepezil significantly upregulated the expression level of BDNF, as well as the phosphorylated level of TrkB. These results suggested that donepezil could protect neurocytes from senility and ameliorate learning and memory impairment in the TS model of AD, which appeared to be through regulating the cholinergic system and inhibiting the BDNF/TrkB-dependent signalling pathway. Moreover, the study underlines the potency of TS to be a novel animal model for research on AD, and it deserves intensive attention.     

Lack of Association Between the TGF-β<Subscript>1</Subscript> Gene and Development of COPD in Asians: A Case–Control Study and Meta-analysis

Abnormalities in the transforming growth factor-β₁ (TGF-β₁) gene are thought to be linked to chronic obstructive pulmonary disease (COPD). We investigated the association between the single nuclear polymorphisms (SNPs) of TGF-β₁ and the risk of COPD in a case–control study and meta-analysis. We genotyped 160 cases and 177 control subjects in a local hospital using the Mass-Array^TM Technology Platform and then tested the association of four SNPs in TGF-β₁ (rs6957, rs1800469, rs2241712, and rs2241718) with COPD. Plasma TGF-β₁ level measurement was performed later. A database covering all papers published up to October 30, 2010, was then reviewed. Statistical analysis was performed using Revman 5.0 and STATA 11.0 software. No association was found between TGF-β₁ gene SNPs and an increased risk of COPD in Asians. By meta-analysis, the link of two polymorphisms, rs1800469 and rs1982073, was investigated in seven and eight studies, respectively, involving 1508 COPD patients and 2608 control subjects. The results showed that there was no significant association between an increased risk of COPD in carriers of the T allele (TT+TC) versus the CC genotype in rs1800469 and rs1982073. In ethnic subgroup analysis, the risk of COPD associated with the rs1800469 T allele was not significantly elevated among Asians. TGF-β₁ gene polymorphisms are not associated with an increased risk of COPD in the Asian population.     

<Emphasis Type="Italic">α</Emphasis>-Lipoic acid increases cardiac glucose oxidation independent of AMP-activated protein kinase in isolated working rat hearts

α-Lipoic acid (ALA) is a naturally occurring enantiomer of lipoic acid and is a cofactor of key metabolic enzyme complexes catalyzing the decarboxylation of α-keto acids. It was recently shown that ALA increases insulin sensitivity by activating AMP-activated protein kinase (AMPK) in skeletal muscle. Also, administration of ALA to obese rats increases insulin-stimulated glucose uptake in the whole body. We investigated the metabolic effects of ALA on isolated working rat hearts. ALA (500μM) stimulated glucose oxidation (157 ± 31 nmol·dry wt^–1·min^–1 in control vs 315 ± 63 nmol·dry wt^–1·min^–1 in ALA-treated, p < 0.05) without affecting glycolysis, lactate oxidation,or palmitate oxidation.Cardiac work was not affected by ALA treatment. The effect of ALA on glucose oxidation was not associated with an activation of AMPK. AMPK activity was 190 ± 14 pmol · mg protein^–1·min^–1 in control vs 190±16 pmol·mg protein–1 · min–1 in ALAtreated hearts. This study shows that ALA stimulates glucose oxidation in isolated working rat hearts independent of AMPK activation.The beneficial effects of ALA treatment in diabetic patients may be at least in part related to its effect on glucose metabolism.     

Regulation of p70<Superscript>S6k</Superscript>, GSK-3β, and calcineurin in rat striated muscle during aging

In this study we compared the content and phosphorylation levels of several molecules believed to regulate muscle hypertrophy and fiber type changes in the extensor digitorum longus (EDL), soleus, diaphragm, and heart of adult (6 months), aged (30 months), and very aged (36 months) Fischer 344 × Brown Norway rats. With aging, the mass of the EDL and soleus decreased significantly (~38% and ~36%, respectively), the diaphragm’s mass remained unchanged while the mass of the heart increased (~35%). Western blotting demonstrated that calcineurin (CnA), the 70-kDa ribosomal S6 kinase (p70^S6k), glycogen synthase kinase-3β (GSK-3β), and the phosphorylated forms of GSK-3β and p70^S6k (p-GSK-3β^Ser9 and p-p70^S6kThr389) were regulated differently with aging and between muscle types. Total p70^S6k, GSK-3β, and p-GSK-3β^Ser9 decreased in the aged-atrophic EDL and soleus while p-p70^S6kThr389 increased. Although total p70^S6k content diminished in the continuously active diaphragm, phosphorylation of p70^S6k remained unchanged. Conversely, the expression of GSK-3β and p-GSK-3β^Ser9 increased in the diaphragm. With aging, the amount of p-p70^S6kThr389 decreased ~56% in the heart while p-GSK-3β ^Ser9 increased ~193%. Interestingly, CnA content remained unchanged in the diaphragm, increased ~204% in the EDL, and decreased ~30% and ~65% with aging in the soleus and heart, respectively. These results indicate remarkable differences in the regulation of molecules thought to govern protein synthesis and changes in contractile protein expression.     

IgG Anti-β<Subscript>2</Subscript> Glycoprotein I Antibodies in Malaysian Patients with Antiphospholipid Syndrome and Systemic Lupus Erythematosus: Prevalence and Clinical Correlations

The aim of this study was to investigate the incidence of IgG anticardiolipin antibody (ACL) and IgG anti-β₂ glycoprotein I antibody (anti-β2GPI) positivity in patients with primary or secondary antiphospholipid syndrome (APS) and systemic lupus erythematosus (SLE), to assess the association between IgG ACL and anti-β2GPI, and the relationship between the presence of ACL and anti-β2GPI with the clinical manifestations of APS. IgG ACL and IgG anti-β2GPI levels were measured in 51 SLE patients, 20 patients with SLE and APS (secondary APS) and 11 primary APS patients using commercially available ELISA kits. Relationships between laboratory data and clinical manifestations of the patients were examined. The incidence of IgG ACL positivity was significantly higher in primary (36.4%) and secondary (40%) APS than in SLE (13.7%) patients (P= 0.02). The incidence of IgG anti-β2GPI positivity was significantly higher in primary (54.5%) and secondary (35%) APS than in SLE (7.8%) patients (P= 0.0006). Mean levels of IgG ACL and anti-β2GPI were significantly higher in the primary and secondary APS than in the SLE patients (P= 0.002 for both). A significant relationship was found between IgG ACL and IgG anti-β2GPI (P= 0.01, R ²= 0.56). There was a significant correlation between the presence of IgG ACL and a history of thrombosis in the combined primary and secondary APS group, but not in SLE patients. In conclusion, in this study IgG ACL and IgG anti-β2GPI are closely related and mean levels of IgG ACL and IgG anti-β2GPI are higher in patients with either primary or secondary APS than in SLE patients. Received: 25 January 2002 / Accepted: 29 April 2002     

Studies of the associations between functional β<Subscript>2</Subscript>-adrenergic receptor variants and obesity,hypertension and type 2 diabetes in 7,808 white subjects

Aims/hypothesis Functional and common Arg16Gly and Gln27Glu polymorphisms have been identified in ADRB2, the gene encoding the β₂-adrenergic receptor. These variants have previously been examined for association with obesity, hypertension and diabetes with inconclusive results. Materials and methods We investigated both of these variants in 7,808 unrelated, middle-aged white people for their association with obesity in a case–control study, quantitative trait analysis and meta-analysis. Moreover, both variants were investigated for their potential influence on measures of hypertension and type 2 diabetes by case–control and quantitative trait analyses. Results The present study did not find consistent evidence for an association of these β₂-adrenergic receptor variants with obesity or hypertension; neither did the quantitative trait analyses show any effect of the variants on obesity-related traits. However, both the Gly allele of the Arg16Gly variant and the Glu allele of the Gln27Glu variant showed nominal association with systolic blood pressure. Furthermore, there was a nominal association of the Arg16 allele frequency and genotype distribution with type 2 diabetes; however, no influence on quantitative biochemical phenotypes related to type 2 diabetes was found. A nominal association of the Arg/Gly genotype with the metabolic syndrome was also observed (p = 0.003). Logistic regression analyses provided no evidence of a synergistic or an additive effect of these variants on obesity, hypertension or diabetes. Conclusions/interpretation After studying 7,808 middle-aged white subjects, we were unable to demonstrate any consistent associations between two common amino acid polymorphisms of the β₂-adrenergic receptor and obesity, hypertension or type 2 diabetes.