Three novel sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) 3 isoforms. Expression, regulation, and function of the membranes of the SERCA3 family

Sarco/endoplasmic reticulum Ca2+-ATPases (SERCAs) pump Ca2+ into the endoplasmic reticulum. Recently, three human SERCA3 (h3a-c) proteins and a previously unknown rat SERCA3 (r3b/c) mRNA have been described. Here, we (i) document two novel human SERCA3 splice variants h3d and h3e, (ii) provide data for the expression and mechanisms regulating the expression of all known SERCA3 variants (r3a, r3b/c, and h3a-e), and (iii) show functional characteristics of the SERCA3 isoforms. h3d and h3e are issued from the insertion of an additional penultimate exon 22 resulting in different carboxyl termini for these variants. Distinct distribution patterns of the SERCA3 gene products were observed in a series of cell lines of hematopoietic, epithelial, embryonic origin, and several cancerous types, as well as in panels of rat and human tissues. Hypertension and protein kinase C, calcineurin, or retinoic acid receptor signaling pathways were found to differently control rat and human splice variant expression, respectively. Stable overexpression of each variant was performed in human embryonic kidney 293 cells, and the SERCA3 isoforms were fully characterized. All SERCA3 isoforms were found to pump Ca2+ with similar affinities. However, they modulated the cytosolic Ca2+ concentration ([Ca2+]c) and the endoplasmic reticulum Ca2+ content ([Ca2+]er) in different manners. A newly generated polyclonal antibody and a pan-SERCA3 antibody proved the endogenous expression of the three novel SERCA3 proteins, h3d, h3e, and r3b/c. All these data suggest that the SERCA3 gene products have a more widespread role in cellular Ca2+ signaling than previously appreciated.     

The Ophthalmic Branch of the Gutenberg Health Study: Study Design,Cohort Profile and Self-Reported Diseases

Purpose

This paper describes the study design, methodology, cohort profile and self-reported diseases in the ophthalmological branch of the Gutenberg Health Study (GHS).

Methods

The GHS is an ongoing, prospective, interdisciplinary, single-center, population-based cohort study in Germany. The main goals of the ophthalmological section are to assess the prevalence and incidence of ocular diseases and to explore risk factors, genetic determinants and associations with systemic diseases and conditions. The eye examination at baseline included a medical history, self-reported eye diseases, visual acuity, refractive errors, intraocular pressure, visual field, pachymetry, keratometry, fundus photography and tear sampling. The 5-year follow-up visit additionally encompassed optical coherence tomography, anterior segment imaging and optical biometry. The general examination included anthropometry; blood pressure measurement; carotid artery ultrasound; electrocardiogram; echocardiography; spirometry; cognitive tests; questionnaires; assessment of mental conditions; and DNA, RNA, blood and urine sampling.

Results

Of 15,010 participants (aged 35-74 years at the time of inclusion), ocular data are available for 14,700 subjects (97.9%). The mean visual acuity (standard deviation), mean spherical equivalent, median decimal visual acuity, and mean intraocular pressure were 0.08 (0.17) logMar, -0.42 (2.43) diopters, 0.9 and 14.24 (2.79) mm Hg, respectively. The frequencies of self-reported strabismus, glaucoma, surgery for retinal detachment and retinal vascular occlusions were 2.7%, 2.3%, 0.2% and 0.4%, respectively.

Conclusions

The GHS is the most extensive dataset of ophthalmic diseases and conditions and their risk factors in Germany and one of the largest cohorts worldwide. This dataset will provide new insight in the epidemiology of ophthalmic diseases and related medical specialties.     

Results of Automated Retinal Image Analysis for Detection of Diabetic Retinopathy from the Nakuru Study,Kenya

Objective

Digital retinal imaging is an established method of screening for diabetic retinopathy (DR). It has been established that currently about 1% of the world’s blind or visually impaired is due to DR. However, the increasing prevalence of diabetes mellitus and DR is creating an increased workload on those with expertise in grading retinal images. Safe and reliable automated analysis of retinal images may support screening services worldwide. This study aimed to compare the Iowa Detection Program (IDP) ability to detect diabetic eye diseases (DED) to human grading carried out at Moorfields Reading Centre on the population of Nakuru Study from Kenya.

Participants

Retinal images were taken from participants of the Nakuru Eye Disease Study in Kenya in 2007/08 (n = 4,381 participants [NW6 Topcon Digital Retinal Camera]).

Methods

First, human grading was performed for the presence or absence of DR, and for those with DR this was sub-divided in to referable or non-referable DR. The automated IDP software was deployed to identify those with DR and also to categorize the severity of DR.

Main Outcome Measures

The primary outcomes were sensitivity, specificity, and positive and negative predictive value of IDP versus the human grader as reference standard.

Results

Altogether 3,460 participants were included. 113 had DED, giving a prevalence of 3.3% (95% CI, 2.7–3.9%). Sensitivity of the IDP to detect DED as by the human grading was 91.0% (95% CI, 88.0–93.4%). The IDP ability to detect DED gave an AUC of 0.878 (95% CI 0.850–0.905). It showed a negative predictive value of 98%. The IDP missed no vision threatening retinopathy in any patients and none of the false negative cases met criteria for treatment.

Conclusions

In this epidemiological sample, the IDP’s grading was comparable to that of human graders’. It therefore might be feasible to consider inclusion into usual epidemiological grading.     

Cytologic diagnosis of intraocular lymphoma in vitreous aspirates

OBJECTIVE: To evaluate the cytologic findings of vitreous fluids with atypical, suspicious for malignancy or malignant lymphoid cells to assess cytologic parameters that may help in reaching the diagnosis of intraoclular lymphoma. STUDY DESIGN: Vitreous aspirates with a malignant, suspicious for malignancy or atypical lymphoid population were identified from the files of Barnes-Jewish Hospital during the previous 11 years. Cytologic preparations were reviewed. Pertinent clinical information was obtained from medical records. RESULTS: Thirteen vitreous aspirates from 12 patients were included. The chief complaints included floaters, blurred vision and decreased visual aculity. Bilateral ocular involvement was present in 8 (67%) patients. Three patients had a history of an extraocular lymphoid malignancy. All patients underwent pars plana vitrectomy and collection of the vitreous aspirate. Cytologic diagnoses included: malignant lymphoma (9 of 13), suspicious for malignant lymphoma (3 of 13) and atypical lymphoid population (1 of 13). Most samples had high cellularity (11 of 13) and necrosis (9 of 13). Abnormal lymphoid cells were large (2-4 times the size of a lymphocyte) and had a high nuclear/cytoplasmic ratio, prominent nucleoli, irregular nuclear contours and a fine to coarse chromatin pattern. All cases with malignant cytology had abundant abnormal lymphoid cells; inconclusive cases had few. Immunocytochemistry for CD20 and CD45RO was performed on 9 of 13 samples and was conclusive in 6 of 9. CONCLUSION: Cytologic analysis of vitreous aspirates can be useful in diagnosing intraocular involvement by malignant lymphoma. Sparse cellularity is the main factor leading to inconclusive diagnoses. Immunostaining can be useful in confirming the lymphoid nature of the malignant cells.     

Bioactive content and antioxidant capacity of Cape gooseberry fruit

At present, Cape gooseberry (Physalis peruviana) fruit is one of the less used raw materials of plant origin, which can be used for human nutrition. This fruit, as well as alimentary products made of it, were used by healers in folk medicine in the distant past. The aim of this study was to monitor and evaluate the antioxidant capacity of fresh fruit of three Cape gooseberry cultivars ‘Giant’, ‘Golden berry’ and ‘Inka’. Antioxidant capacity was also tested, on the basis of the scavenging effect of reactive oxygen species (ROS) and lipid peroxidation of methanolic extracts made of fresh fruit. These results were further extended and supplemented with determinates of the vitamin C and total phenolic contents. These analyses were made for three consecutive years. The highest values of antioxidant capacity were observed in the ‘Inka’ cultivar (9.31 grams of ascorbic acid equivalents kg⁻¹ of fresh mass). In this cultivar, the obtained results were corroborated also in ROS and the contents of vitamin C and total phenolics. Due to a high antioxidant capacity of this fruit species, the results presented should increase its popularity above all as a promising raw material, which can be used for human nutrition.     

Retinal Arterioles Narrow with Increasing Duration of Anti-Retroviral Therapy in HIV Infection: A Novel Estimator of Vascular Risk in HIV?

Objectives

HIV infection is associated with an increased risk of age-related morbidity mediated by immune dysfunction, atherosclerosis and inflammation. Changes in retinal vessel calibre may reflect cumulative structural damage arising from these mechanisms. The relationship of retinal vessel calibre with clinical and demographic characteristics was investigated in a population of HIV-infected individuals in South Africa.

Methods

Case-control study of 491 adults ≥30 years, composed of 242 HIV-infected adults and 249 age- and gender-matched HIV-negative controls. Retinal vessel calibre was measured using computer-assisted techniques to determine mean arteriolar and venular diameters of each eye.

Results

The median age was 40 years (IQR: 35–48 years). Among HIV-infected adults, 87.1% were receiving highly active antiretroviral therapy (HAART) (median duration, 58 months), their median CD4 count was 468 cells/µL, and 84.3% had undetectable plasma viral load. Unadjusted mean retinal arteriolar diameters were 163.67±17.69 µm in cases and 161.34±17.38 µm in controls (p = 0.15). Unadjusted mean venular diameters were 267.77±18.21 µm in cases and 270.81±18.98 µm in controls (p = 0.07). Age modified the effect of retinal arteriolar and venular diameters in relation to HIV status, with a tendency towards narrower retinal diameters in HIV cases but not in controls. Among cases, retinal arteriolar diameters narrowed with increasing duration of HAART, independently of age (167.83 µm <3 years of HAART vs. 158.89 µm >6 years, p-trend = 0.02), and with a HIV viral load >10,000 copies/mL while on HAART (p = 0.05). HIV-related venular changes were not detected.

Conclusions

Narrowing of retinal arteriolar diameters is associated with HAART duration and viral load, and may reflect heightened inflammatory and pro-atherogenic states of the systemic vasculature. Measurement of retinal vascular calibre could be an innovative non-invasive method of estimating vascular risk in HIV-infected individuals.     

Optimal strategy for controlling the spread of HIV/AIDS disease: a case study of South Africa

HIV/AIDS disease continues to spread alarmingly despite the huge amounts of resources invested in fighting it. There is a need to integrate the series of control measures available to ensure a consistent reduction in the incidence of the disease pending the discovery of its cure. We present a deterministic model for controlling the spread of the disease using change in sexual habits and antiretroviral (ARV) therapy as control measures. We formulate a fixed time optimal control problem subject to the model dynamics with the goal of finding the optimal combination of the two control measures that will minimize the cost of the control efforts as well as the incidence of the disease. We estimate the model state initial conditions and parameter values from the demographic and HIV/AIDS data of South Africa. We use Pontryagin's maximum principle to derive the optimality system and solve the system numerically. Compared with the practice in most resource-limited settings where ARV treatment is given only to patients with full-blown AIDS, our simulation results suggest that starting the treatment as soon as the patients progress to the pre-AIDS stage of the disease coupled with appreciable change in the susceptible individuals' sexual habits reduces both the incidence and prevalence of the disease faster. In fact, the results predict that the implementation of the proposed strategy would drive new cases of the disease towards eradication in 10 years.     

人β干扰素N端序列的变异与其抗病毒活性的关系

研究了人β干扰素N端序列的变异与其抗病毒活性的关系。采用DNA重组技术获得了两种干扰素变种:一种N端序列为MWIRGF,另一种为MAIRGF。这两种干扰素变种的抗病毒活性都明显下降。此结果提示,人β干扰素的N端序列对抗病毒活性起主导作用,在改造该蛋白结构时,应避免改动N端结构。     

Centromere proteins

Human anti-centromere sera from scleroderma patients were used to detect centromere antigens of mouse fibroblast cells. An M_r=59000 centromere protein was localized exclusively on mitotic chromosomes. The association of this protein with the mitotic chromosomes proved to be DNase I sensitive. In interphase nuclei, this centromere antigen was not detectable by immunoblot techniques. The results suggest that the M_r=59000 mitosis specific protein may be necessary for the structural stability of kinetochores during mitosis.