β1-Adrenoceptor gene variations: a role in idiopathic dilated cardiomyopathy?

A substantial body of evidence suggests involvement of the human beta1-adrenoceptor (beta1-AR) gene in the pathophysiology of dilated cardiomyopathy (DCM), a severe heart disease of significant public health impact. Beta1-AR-mediated signal transduction is dramatically altered due to downregulation, resulting in an impairment of myocardial response. The important role of genetic factors in idiopathic dilated cardiomyopathy (IDCM) recently recognized, we analyzed this prime candidate gene for genetic variation in carefully selected patients and controls. In this preliminary study, 18 single nucleotide polymorphisms were observed, 17 of which were located in the N-terminal and C-terminal region of the coding exon, resulting in 7 amino acid exchanges: Ser-49-Gly, Ala-59-Ser, Gly-389-Arg, Arg-399-Cys, His-402-Arg, Thr-404-Ala, and Pro-418-Ala. These mutations resulted in 11 different beta1-AR genotypes. Importantly, the genotypes carrying the Ser-49-Gly mutation in the N-terminus of the molecule in a heterozygous or homozygous form were observed significantly more frequently in the group of IDCM patients. The present results may provide a clue on the molecular mechanisms involved in IDCM, and add moreover interesting information on nature, distribution, and evolutionary aspects of sequence variation in human adrenergic receptor genes.     

Our surgical heritage. Surgery in ancient Egypt

In its 3,000 years History the surgery of the Old Egypt came on to an important development. Some of the antique instruments used in traumatology, the general surgery and in cosmetic-plastic operations, are in a scarcely modified manner employed for the same purposes in modern surgical interventions nowadays. The surgical diagnostics and therapy of that time is demonstrated by the surgical instruments stock being in the possession of the "Agyptisches Museum" of the "Staatlichen Museen zu Berlin". The surgery of Egypt transferred its leading part to the prosperous medical schools of Greece at the late period of the Old Egyptian empire (1085-332 B.C.). Its surgical diagnostics and therapy depending on an empirical rationalism, the applied instruments but also its ethical attitude towards the patients have been one of the dedisive bases influencing the development of the surgery in the antiquity.     

Enolase 1 of Candida albicans binds human CD4+ T cells and modulates naïve and memory responses

To obtain a better understanding of the biology behind life-threatening fungal infections caused by Candida albicans, we recently conducted an in silico screening for fungal and host protein interaction partners. We report here that the extracellular domain of human CD4 binds to the moonlighting protein enolase 1 (Eno1) of C. albicans as predicted bioinformatically. By using different anti-CD4 monoclonal antibodies, we determined that C. albicans Eno1 (CaEno1) primarily binds to the extracellular domain 3 of CD4. Functionally, we observed that CaEno1 binding to CD4 activated lymphocyte-specific protein tyrosine kinase (LCK), which was also the case for anti-CD4 monoclonal antibodies tested in parallel. CaEno1 binding to naïve human CD4⁺ T cells skewed cytokine secretion toward a Th2 profile indicative of poor fungal control. Moreover, CaEno1 inhibited human memory CD4⁺ T-cell recall responses. Therapeutically, CD4⁺ T cells transduced with a p41/Crf1-specific T-cell receptor developed for adoptive T-cell therapy were not inhibited by CaEno1 in vitro. Together, the interaction of human CD4⁺ T cells with CaEno1 modulated host CD4⁺ T-cell responses in favor of the fungus. Thus, CaEno1 mediates not only immune evasion through its interference with complement regulators but also through the direct modulation of CD4⁺ T-cell responses.     

Identifying children with high mortality risk

This issue reports selected results from a comprehensive study of infant and child mortality based on the National Family Health Survey data. The analysis distinguishes between neonatal, postneonatal, infant and child mortality since mortality and its causes vary considerably among children of different ages. Hazard regression analysis was used to estimate the effects of each individual variable as the factors that affect infant and child mortality tend to be correlated with each other. The study involves adjusted effects of selected socioeconomic and demographic characteristics on neonatal, postneonatal, infant, and child mortality for children born during the survey. Short birth intervals have a great effect on infant and child mortality. A previous birth interval of less than 24 months increases child mortality by about 67%. Neonatal mortality is highest among children of very young mothers. Child mortality is higher for girls in all states except Tamil Nadu, Kerala, and Goa. Seven groups of children who are especially vulnerable to infant and child mortality were identified. Thus, intervention programs, such as efforts to provide supplemental nutrition and basic immunization to pregnant mothers, infants and young children need to focus on these high-risk groups. Results for many states show elevated mortality rates for girls after the neonatal period. Family health programs aimed at overall improvement in mortality levels should pay attention to providing basic health care and supplemental nutrition to girls.     

Magnetic Resonance Angiography of the Abdominal Aorta

Magnetic resonance angiography (MRA) of the abdominal aorta was performed in 36 patients using the "bright-blood" technique on the basis of a series of flow-compensated breath-hold 2 D-GE pulse sequences. Diverse diseases included complete occlusion, stenosis, atherosclerotic or dissecting aneurysm and renal artery stenosis. Coronal and axial projection angiograms [maximum intensity projection (MIP) algorithm] and individual GE images were compared with DSA and contrast-enhanced computed tomography (CT). Our data showed a good correlation of MR and digital subtraction arteriography (DSA) or CT resp. in all cases, when both rotating MIP angiograms and individual GE images were analyzed. Thus, MR is suggested to be a useful noninvasive diagnostic method for the abdominal aorta, particularly in preoperative staging of aneurysms, aortic occlusive disease, and stenosis of the proximal main renal artery. Drawbacks of the method are a signal loss in slow or turbulent flow conditions, and unsatisfactory spatial resolution in small vessels.     

Surgical treatment of 50 carotid dissections: indications and results

PURPOSE: This article analyzes the course of 48 patients with 49 chronic carotid dissections (who were treated surgically at our institution after a median anticoagulation period of 9 months because of a persistent high-grade stenosis or an aneurysm) and the course of one additional patient with acute carotid dissection (who underwent early operative reconstruction 12 hours after onset because of fluctuating neurologic symptoms). METHODS: All medical and surgical records and imaging studies were reviewed retrospectively. All histologic specimens were reevaluated by a single pathologist to assess the cause of dissection. Follow-up of 41 patients (85%) after 70 months (range, 1-190 months) consisted of an examination of the extracranial vessels in the neck by Doppler ultrasound scanning and a questionnaire about the patients' medical history and their personal appraisals of cranial nerve function. RESULTS: Seventy percent of the dissections had developed spontaneously; 18% were caused by trauma; 12% of all patients (22% of the women) had a fibromuscular dysplasia. Indication for surgery was a high-grade persisting stenosis and a persisting or newly developed aneurysm. Flow restoration was achieved by resection and vein graft replacement in 40 cases (80%) and thromboendarterectomy and patch angioplasty in three cases (6%). Gradual dilatation was performed and effective in two cases (4%). Five internal carotid arteries (10%) had to be clipped because dissection extended into the skull base. One patient died of intracranial bleeding. Five patients (10%) experienced the development of a recurrent minor stroke (ipsilateral, 4 patients; contralateral, 1 patient). Cranial nerve damage could not be avoided in 29 cases (58%) but were transient in most of the cases. During follow-up, one patient died of unrelated reasons, and only one patient had experienced the development of a neurologic event of unknown cause. CONCLUSION: Chronic carotid dissection can be effectively treated by surgical reconstruction to prevent further ischemic or thromboembolic complications, if medical treatment for 6 months with anticoagulation failed or if carotid aneurysms and/or high-grade carotid stenosis persisted or have newly developed.     

Hispanic health status in Orange County, Florida.

PURPOSE: To assess the health status of the Hispanic population of Orange County, Florida. METHODS: The methodology utilized secondary data for 66 ethnically identified indicators in a comparative framework applied for a 5-year period (1997-2001). FINDINGS: Orange County Hispanics are younger with lower per capita income than their Florida peers, less likely to be White, and much more likely to be of Puerto Rican origin. Relative to the Hispanic populations in the selected peer counties and statewide, Orange County Hispanics have higher age-adjusted death rates for a majority of disease categories and conditions, such as breast, lung, and prostate cancers; chronic liver disease and cirrhosis; diabetes mellitus; pneumonia and influenza; stroke; acquired immunodeficiency syndrome; motor vehicle accidents; and infant, neonatal, and child mortality. Orange County Hispanics did better in comparison to Orange non-Hispanics, with lower age-adjusted death rates for major causes of death such as heart disease, cancer, and stroke. However, for many indicators, the 5-year trends for Orange County Hispanics are moving in an unfavorable direction in contrast to the trends for non-Hispanics, which are either stable or improving. CONCLUSION: Comparative assessments of Hispanic populations using secondary data enable the development of a comprehensive health status profile. However, this approach is currently constrained by the limited number of ethnically identified indicators and, especially for Hispanics, problems in the accuracy and consistency of the assignment to racial categories and subsequent reporting.     

Expression and characterization of Gag protein of HIV-1（CN） in Pichia pastoris

To express the core protein of HIV-1 of Chinese prevalent strain （HIV-1 （CN）） in Pichia pastoris, the fulllength gag gene was inserted into the secretory expression vector pHILS1. Linearized recombinant plasmid pHILGAG by Sail was electrotransformed into the yeast strain GS115, and the yeast transformants were identified by PCR. To induce the interest protein to be expressed, the PCR positive transformants were inoculated in the medium of BMGY and BMMY, mRNA of the strain was detected by RT-PCR, and the expressed protein was analyzed by SDS-PAGE, Western blotting and thin layer scanning. mRNA （1.3 kb） was amplified by RT-PCR. SDS-PAGE and Western blotting analysis showed that the molecular mass of the expressed protein was 55 kD, which was similar to the expected value, and the expressed protein could react with McAb to HIV-1 p24. Thin layer scanning analysis demonstrated that the whole amount of the expressed protein was approximately 13 % of the soluble protein in the supernatant. The recombinant yeast had good genetic stability. The optimal expression conditions of the engineering yeast were as follows： BMMY medium, 80-90% of dissolved oxygen, 1% methanol, and 3-day-cultivation course. Gag proteins were expressed under the optimal expression condition and purified via gel filtration chromatography. The purity of the interest protein was up to 85 %. After the purified proteins were inoculated into BALB/c mice, the anti-HIV-1 antibodies in the immunized mice could be detected by Western blotting.     

Lesion-associated accumulation of uPAR/CD87- expressing infiltrating granulocytes, activated microglial cells/macrophages and upregulation by endothelial cells following TBI and FCI in humans

Urokinase-type plasminogen activator receptor (uPAR/CD87) together with its ligand, urokinase-type plasminogen activator (uPA), constitutes a proteolytic system associated with tissue remodelling and leucocyte infiltration. uPAR is a member of the glycosyl phosphatidyl inositol (GPI) anchored protein family. The functional role of uPAR comprises fibrinolysis by conversion of plasminogen to plasmin. In addition, uPAR promotes cell adhesion, migration, proliferation, re-organization of the actin cytoskeleton, and angiogenesis. Furthermore, uPAR is involved in prevention of scar formation and is chemoattractant to macrophages and leucocytes. In order to investigate the pathophysiological role of uPAR following human CNS injury we examined necrotic brain lesions resulting from traumatic brain injury (TBI; n = 28) and focal cerebral infarctions (FCI; n = 17) by immunohistochemistry. Numbers of uPAR+ cells and uPAR+ blood vessels were counted. Following brain damage, uPAR+ cells increased significantly within 12 h, reached a maximum after 3-4 days and remained elevated until later stages. uPAR was expressed by infiltrating granulocytes, activated microglia/macrophages and endothelial cells. Numbers of uPAR+ vessels increased in parallel subsiding earlier following FCI than post TBI. The restricted, lesion-associated accumulation of uPAR+ cells in the brain parenchyma and upregulated expression by endothelial cells suggests a crucial role for the influx of inflammatory cells and blood-brain barrier (BBB) disturbance. Through a failure in BBB function, uPAR participates in formation of brain oedema and thus contributes to secondary brain damage. In conclusion, the study defines the localization, kinetic course and cellular source of uPAR as a potential pharmacological target following human TBI and FCI.