Apolipoprotein E gene polymorphism and the risk of left ventricular dysfunction among Egyptian β-thalassemia major

In Egypt, β-thalassemia is the most common hereditary hemolytic anemia. Cardiac dysfunction, secondary to iron overload with formation of oxygen free radicals, is the most common cause of death in β-thalassemia patients. This study was designed to determine whether the allelic genotype of apolipoprotein E (Apo E), which exhibits antioxidant properties, could represent a genetic risk factor for the development of left ventricular (LV) dysfunction in β-thalassemia major. Fifty Egyptian β-thalassemia major patients were subjected to echocardiography to assess LV function. Apo E genotyping by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) was done for all patients in addition to 50 age and sex matched healthy control subjects. Patients were classified into three groups. Group I and II were clinically asymptomatic. Group II subjects had evidence of LV dilatation, while Group III patients had clinical and echocardiographic findings of LV failure. Apo E4 allele was significantly higher among Group II and III than in controls. In conclusion, Apo E4 allele can be considered as a genetic risk factor for LV dysfunctions in β-thalassemic patients. It could be used as predictive indicator for additional risk of LV failure, particularly in asymptomatic patients with LV dilatation, requiring a closer follow-up, to prevent further disease progression.     

Assessment of HER-2 gene overexpression in Isfahan province breast cancer patients using Real Time RT-PCR and immunohistochemistry

Purpose

Overexpression of proto-oncogene HER-2 is one of the main molecular markers of breast cancer involved in prognosis and diagnosis and also in trastuzumab therapy. Thus, a request for the evaluation of HER-2 status in breast cancer has been increasing. The aim of our study was assessment of HER-2 overexpression in malignant and benign breast cancer specimens by Real Time RT-PCR technique and comparison of its results with IHC outcomes.

Methods

Twenty benign and sixty malignant breast cancers in addition to fifteen normal breast tissue specimens were analyzed by Real Time RT-PCR method. Fresh tissue samples were disrupted by mortar and pestle. A syringe and a needle were used for complete homogenization of the tissues. The RNA was then isolated from the samples and converted to cDNA. A standard curve was initially plotted using BioEasy SYBR Green I and then all 95 specimens were studied by Real Time RT-PCR using 2^− ΔΔCt method.

Results

23.3% of 60 malignant specimens showed HER-2 overexpression, while all of the benign samples represented the normal expression level of HER-2 gene. The concordance rate between the results of Real Time RT-PCR and IHC was 86.6%.

Conclusion

Real Time RT-PCR method is an almost reliable technique and at least can be used as a complementary method for confirming IHC results. This is emanated from relatively high rate of concordance between outcomes of IHC test, as a routine method of detecting the HER-2 gene expression status, and Real Time RT-PCR technique.     

Irreversible Heavy Chain Transfer to Hyaluronan Oligosaccharides by Tumor Necrosis Factor-stimulated Gene-6

The covalent transfer of heavy chains (HCs) from inter-α-inhibitor (IαI) to hyaluronan (HA) via the protein product of tumor necrosis factor-stimulated gene-6 (TSG-6) forms the HC-HA complex, a pathological form of HA that promotes the adhesion of leukocytes to HA matrices. The transfer of HCs to high molecular weight (HMW) HA is a reversible event whereby TSG-6 can shuffle HCs from one HA molecule to another. Therefore, HMW HA can serve as both an HC acceptor and an HC donor. In the present study, we show that transfer of HCs to low molecular weight HA oligosaccharides is an irreversible event where subsequent shuffling does not occur, i.e. HA oligosaccharides from 8 to 21 monosaccharide units in length can serve as HC acceptors, but are unable to function as HC donors. We show that the HC-HA complex is present in the synovial fluid of mice subjected to systemic and monoarticular mouse models of rheumatoid arthritis. Furthermore, we demonstrate that HA oligosaccharides can be used, with TSG-6, to irreversibly shuffle HCs from pathological, HMW HC-HA to HA oligosaccharides, thereby restoring HC-HA matrices from the inflamed joint to their normal state, unmodified with HCs. This process was also effective for HC-HA in the synovial fluid of human rheumatoid arthritis patients (in vitro).     

A Shared Endoplasmic Reticulum-associated Degradation Pathway Involving the EDEM1 Protein for Glycosylated and Nonglycosylated Proteins

Studies of misfolded protein targeting to endoplasmic reticulum-associated degradation (ERAD) have largely focused on glycoproteins, which include the bulk of the secretory proteins. Mechanisms of targeting of nonglycosylated proteins are less clear. Here, we studied three nonglycosylated proteins and analyzed their use of known glycoprotein quality control and ERAD components. Similar to an established glycosylated ERAD substrate, the uncleaved precursor of asialoglycoprotein receptor H2a, its nonglycosylated mutant, makes use of calnexin, EDEM1, and HRD1, but only glycosylated H2a is a substrate for the cytosolic SCF^Fbs2 E3 ubiquitin ligase with lectin activity. Two nonglycosylated BiP substrates, NS-1κ light chain and truncated Igγ heavy chain, interact with the ERAD complex lectins OS-9 and XTP3-B and require EDEM1 for degradation. EDEM1 associates through a region outside of its mannosidase-like domain with the nonglycosylated proteins. Similar to glycosylated substrates, proteasomal inhibition induced accumulation of the nonglycosylated proteins and ERAD machinery in the endoplasmic reticulum-derived quality control compartment. Our results suggest a shared ERAD pathway for glycosylated and nonglycosylated proteins composed of luminal lectin machinery components also capable of protein-protein interactions.     

hCOA3 Stabilizes Cytochrome c Oxidase 1 (COX1) and Promotes Cytochrome c Oxidase Assembly in Human Mitochondria

Cytochrome c oxidase (COX) or complex IV of the mitochondrial respiratory chain plays a fundamental role in energy production of aerobic cells. In humans, COX deficiency is the most frequent cause of mitochondrial encephalomyopathies. Human COX is composed of 13 subunits of dual genetic origin, whose assembly requires an increasing number of nuclear-encoded accessory proteins known as assembly factors. Here, we have identified and characterized human CCDC56, an 11.7-kDa mitochondrial transmembrane protein, as a new factor essential for COX biogenesis. CCDC56 shares sequence similarity with the yeast COX assembly factor Coa3 and was termed hCOA3. hCOA3-silenced cells display a severe COX functional alteration owing to a decreased stability of newly synthesized COX1 and an impairment in the holoenzyme assembly process. We show that hCOA3 physically interacts with both the mitochondrial translation machinery and COX structural subunits. We conclude that hCOA3 stabilizes COX1 co-translationally and promotes its assembly with COX partner subunits. Finally, our results identify hCOA3 as a new candidate when screening for genes responsible for mitochondrial diseases associated with COX deficiency.     

Discovery of a potent respiratory syncytial virus RNA polymerase inhibitor

Targeting viral polymerases has been a proven and attractive strategy for antiviral drug discovery. Herein we describe our effort in improving the antiviral activity and physical properties of a series of benzothienoazepine compounds as respiratory syncytial virus (RSV) RNA polymerase inhibitors. The antiviral activity and spectrum of this class was significantly improved by exploring the amino substitution of the pyridine ring, resulting in the discovery of the most potent RSV A polymerase inhibitors reported to date.     

脉冲震荡肺功能在稳定期COPD治疗中的应用价值

目的:探讨脉冲震荡肺功能(Impulse oscillometry,IOS)在稳定期COPD患者中应用价值及其和常规肺功能检测指标的相关性。方法:62例重度稳定期COPD患者,同时选择健康对照组人群40例纳入研究。经噻托溴铵联合沙美特罗替卡松治疗,将重度COPD患者缓解至中度。检测治疗前后常规肺功能指标(FEV1/FVC、FEV1)和IOS指标(ZRS、Fres、R5、X5、R20),分析常规肺功能指标和IOS各指标的相关性。结果:COPD患者FEV1和FEV1/FVC和对照组相比明显降低,差异有统计学意义(P0.05),观察组治疗后FEV1和FEV1/FVC明显改善,差异有统计学意义(P0.05)。观察组ZRS、Fres、R5、R20各项指标明显高于对照组,差异有统计学意义(P0.05);治疗后ZRS、Fres、R5明显改善,差异有统计学意义(P0.05)。FEV1、FEV1/FVC和ZRS、Fres、R5呈负相关性(P0.05),和X5呈正相关性(P0.05)。结论:脉冲震荡肺功能多项指标和传统肺功能指标有良好的相关性,是一种简便、低配合度、准确的的肺功能新的检测技术手段。     

血清PCT联合痰培养检测在呼吸道感染疾病中的应用

目的:研究血清降钙素原(PCT)联合痰培养检测在呼吸道感染疾病中的应用价值。方法:选择2012年1月至2014年12月在我院接受治疗的呼吸道感染患者100例进行研究。分析痰培养菌群的分布情况以及PCT的检测结果,对比痰培养细菌感染者及真菌感染者的菌种分类及PCT检测结果。结果:检出病原菌的患者PCT水平均分别显著高于正常菌群及未见细菌生长的患者,差异均有统计学意义(均P0.05)。细菌感染PCT阳性比例与真菌感染相比,差异无统计学意义(P0.05)。细菌感染中,革兰氏阳性菌的PCT水平(0.74±0.33μg/L)与革兰氏阴性菌(0.72±0.24μg/L)对比,差异无统计学意义(P0.05)。真菌感染中,白假丝酵母菌及烟曲霉菌的PCT阳性比例最高,分别为60.00%及80.00%。结论:血清PCT与痰培养检测应用于评价有呼吸道感染的患者病情,具有一定的反馈意义,但无法确定患者的呼吸道感染究竟是细菌感染亦或是真菌感染。     

纤维支气管镜对急诊重症肺炎合并呼吸衰竭患者呼吸功能及血清炎症因子水平的影响

目的:探讨纤维支气管镜对急诊重症肺炎合并呼吸衰竭患者呼吸功能及炎症反应的影响。方法:选择2013年9月-2015年9月在我院接受治疗的重症肺炎合并呼吸衰竭患者89例作为研究对象,根据治疗方法不同,将患者分为研究组(47例)与对照组(42例)。对照组患者采用常规抗感染治疗,研究组患者在对照组基础上采用纤维支气管镜肺泡灌洗治疗。观察并比较两组患者治疗前后的动态顺应性(Cdyn)、氧合指数(Pa O2/Fi O2)、呼吸做功(WOB)、血清炎症因子水平的变化情况。结果:治疗前,两组患者Cdyn,WOB,Pa O2/Fi O2,CD11b+中性粒细胞、STREM-1及HMGB-1水平比较,差异均无统计学意义(P0.05);治疗后,对照组患者Cdyn、Pa O2/Fi O2及WOB均较治疗前显著升高,差异具有统计学意义(P0.05);研究组患者Cdyn及Pa O2/Fi O2较治疗前显著升高,而WOB较治疗前显著降低,差异具有统计学意义(P0.05);研究组患者Cdyn及Pa O2/Fi O2高于对照组,而WOB低于对照组,差异具有统计学意义(P0.05);两组患者CD11b+中性粒细胞、STREM-1及HMGB-1水平均较治疗前显著降低,且研究组显著低于对照组,差异均具有统计学意义(P0.05)。结论:纤维支气管镜肺泡灌洗能够改善急诊重症肺炎合并呼吸衰竭患者的呼吸功能,缓解机体炎症反应,具有重要的临床意义。