Retinal gene therapy with a large MYO7A cDNA using adeno-associated virus.

Usher 1 patients are born profoundly deaf and then develop retinal degeneration. Thus they are readily identified before the onset of retinal degeneration, making gene therapy a viable strategy to prevent their blindness. Here, we have investigated the use of adeno-associated viruses (AAVs) for the delivery of the Usher 1B gene, MYO7A, to retinal cells in cell culture and in Myo7a-null mice. MYO7A cDNA, under control of a smCBA promoter, was packaged in single AAV2 and AAV5 vectors and as two overlapping halves in dual AAV2 vectors. The 7.9-kb smCBA-MYO7A exceeds the capacity of an AAV vector; packaging of such oversized constructs into single AAV vectors may involve fragmentation of the gene. Nevertheless, the AAV2 and AAV5 single vector preparations successfully transduced photoreceptor and retinal pigment epithelium cells, resulting in functional, full-length MYO7A protein and correction of mutant phenotypes, suggesting successful homologous recombination of gene fragments. With discrete, conventional-sized dual AAV2 vectors, full-length MYO7A was detected, but the level of protein expression was variable, and only a minority of cells showed phenotype correction. Our results show that MYO7A therapy with AAV2 or AAV5 single vectors is efficacious; however, the dual AAV2 approach proved to be less effective

Excessive Hospitalizations and Its Associated Economic Burden among People with Diabetes in the United States

AbstractObjectivesWe conducted this study to estimate the excessive hospital admission among people with diabetes and the associated economic burden in the USA.MethodsThe study was based on the 2005 Nationwide Inpatient Sample (NIS), a nationally representative probability sample conducted annually by the Agency of Health Research and Quality. Nearly 8 million records were sampled from over 1000 community hospitals in the 2005 NIS.ResultsExcluding childbirth-related admissions, the estimated US hospitalizations numbered approximately 30.8 million; individuals with diabetes accounted for over 6.4 million (20.9%) of these admissions. For every 1000 individuals without diabetes, with type 1 diabetes, and with type 2 diabetes, the numbers of hospitalizations were 89, 418, and 303, respectively. The rates of hospitalization increased greatly by the presence of diabetes for all age groups and sex. During 2005, the national bill of hospital charges and costs for individuals with diabetes exceeded US$171 billion and US$90 billion, respectively. If the prevalence of diabetes increases to 7.5% from 7.0%, the total number of hospitalizations made by individuals with diabetes will be 7.5 million in 2015.ConclusionsAlthough approximately 7% of the population had diabetes in the USA, nearly 20.9% of hospitalizations were made by individuals with this condition. Due to the excessive hospitalizations incurred by patients with diabetes, a small increase in the number of people with diabetes will amplify the number of hospitalizations. Health-care communities should anticipate this possible increased demand of hospitalizations and the associated economic burden

Frequency Dependent Specific Heat from Thermal Effusion in Spherical Geometry

We present a novel method of measuring the frequency dependent specific heat at the glass transition applied to 5-polyphenyl-4-ether. The method employs thermal waves effusing radially out from the surface of a spherical thermistor that acts as both a heat generator and thermometer. It is a merit of the method compared to planar effusion methods that the influence of the mechanical boundary conditions are analytically known. This implies that it is the longitudinal rather than the isobaric specific heat that is measured. As another merit the thermal conductivity and specific heat can be found independently. The method has highest sensitivity at a frequency where the thermal diffusion length is comparable to the radius of the heat generator. This limits in practise the frequency range to 2-3 decades. An account of the 3omega-technique used including higher order terms in the temperature dependency of the thermistor and in the power generated is furthermore given.Comment: 17 pages, 15 figures, Substantially revised versio

rAAV2/5 Gene-Targeting to Rods: Dose-Dependent Efficiency and Complications Associated With Different Promoters

A prerequisite for using corrective gene therapy to treat humans with inherited retinal degenerative diseases that primarily affect rods is to develop viral vectors that target specifically this population of photoreceptors. The delivery of a viral vector with photoreceptor tropism coupled with a rod-specific promoter is likely to be the safest and most efficient approach to target expression of the therapeutic gene to rods. Three promoters that included a fragment of the proximal mouse opsin promoter (mOP), the human G-protein-coupled receptor protein kinase 1 promoter (hGRK1), or the cytomegalovirus immediate early enhancer combined with the chicken β actin proximal promoter CBA were evaluated for their specificity and robustness in driving GFP reporter gene expression in rods, when packaged in a recombinant adeno-associated viral vector of serotype 2/5 (AAV2/5), and delivered via subretinal injection to the normal canine retina. Photoreceptor-specific promoters (mOP, hGRK1) targeted robust GFP expression to rods, whereas the ubiquitously expressed CBA promoter led to transgene expression in the retinal pigment epithelium, rods, cones and rare Müller, horizontal and ganglion cells. Late onset inflammation was frequently observed both clinically and histologically with all three constructs when the highest viral titers were injected. Cone loss in the injected regions of the retinas that received the highest titers occurred with both the hGRK1 and CBA promoters. Efficient and specific rod transduction, together with preservation of retinal structure was achieved with both mOP and hGRK1 promoters when viral titers in the order of 1011 vg ml–1 were used

Schistosoma haematobium co-infection with soil-transmitted helminthes: prevalence and risk factors from two communities in the central region of Ghana

Background: Schistosoma haematobium co-infection with S. mansoni and  soil-transmitted helminthes afflict most-at-risk populations in endemic communities in the developing world. Aim: This study investigated S. haematobium co-infection with soil-transmitted helminthes, and host risk factors in two communities in the central region of Ghana. Methods: Schools and children were selected by stratified cluster and random sampling methods respectively. A total of 250 school children (aged 5 - > 20 years) were recruited. Teachers issued questionnaires to obtain information on host risk factors, water-contact activities and knowledge of S.  haematobium infection. Urine and stool samples were examined for S. haematobium infection and S. mansoni and soil-transmitted helminthes using sedimentation quantitative and direct smear/formol-ether sedimentation concentration techniques respectively.Results: S. haematobium infection (1 - 50 eggs/10 ml urine)  prevalence at Apewosika and Putubiw were 27.5 % and 17 % respectively. Males were more at risk of S. haematobium infection than females. S. haematobium co-infection with soil-transmitted helminthes (A. lumbricoides, E. histolytica, and T. trichuria) was recorded in Putubiw, with females more at risk than males. Children aged 16-19 and 10-15 were more at risk of S. haematobium infection and  helminthic co-infection respectively. Haematuria and proteinuria were predictive of S. haematobium infection. School children had poor knowledge of S. haematobium infection. Water-contact activity was common.Conclusion: S. haematobium prevalence and its co-infection with soil-transmitted helminthes were common in Putubiw. Watercontact activity and poor knowledge about S. haematobium infection were major risk factors. Increased education on preventive and control measures especially in schools is recommended.Key words: Haematuria, proteinuria, Schistosoma haematobium, S. mansoni, helminthes, Bulinus globosu

The longitudinal link between visual acuity and health-related quality of life in patients with diabetic retinopathy

<p>Abstract</p> <p>Background</p> <p>This study characterized the degree of change in health-related quality of life (HRQL) associated with change in visual acuity among patients with diabetic retinopathy.</p> <p>Methods</p> <p>Data are from a randomized, placebo-controlled trial of ruboxistaurin for vision loss in patients with diabetic retinopathy. Visual acuity was quantified as letters on the ETDRS visual acuity chart. HRQL was assessed with the 25-Item Visual Function Questionnaire (VFQ-25) and the SF-36. Patients were categorized into groups based on visual acuity change from baseline to month 18. HRQL change of these groups was compared using general linear models. Regression analyses examined visual acuity change defined continuously.</p> <p>Results</p> <p>Patients (N = 535) were primarily Caucasian (81.9%) and male (64.1%); mean age = 59.3 years. Compared to patients whose visual acuity did not change, the group with > 10 letters vision loss had significantly greater decreases in all VFQ-25 subscales except ocular pain. SF-36 change scores did not correspond as closely to change in vision. Change in visual acuity defined continuously was significantly associated with change in all VFQ-25 scales except ocular pain (p < 0.0001).</p> <p>Conclusion</p> <p>Change in visual acuity was associated with corresponding changes in HRQL among patients with diabetic retinopathy. Previous research has often defined vision loss as a loss of at least 15 letters on the ETDRS visual acuity chart. In the current study, however, a loss of at least 10 letters was associated with substantial declines in HRQL domains such as driving, dependency, role limitations, and mental health. These findings suggest that patients who experience vision loss of at least 10 letters may be appropriate targets of future research and clinical intervention.</p

Effet de l’eau oxygénée (H2O2) sur la germination et la croissance de la roselle (Hibiscus sabdariffa L. var. sabdariffa) au Gabon

Les quantit&#233;s de roselle (Hibiscus sabdariffa L. var. sabdariffa) produites au Gabon ne sont pas suffisantes pour satisfaire la demande locale. Pour augmenter les productions de cette plante, un essai d&#8217;intensification des rendements a &#233;t&#233; r&#233;alis&#233; en utilisant de l&#8217;eau oxyg&#233;n&#233;e (H2O2). Les graines de roselle ont pour cela &#233;t&#233; trait&#233;es par trempage pendant 48 h dans diff&#233;rentes concentrations de ce compos&#233;, puis cultiv&#233;es en serre. Les r&#233;sultats obtenus ont montr&#233; qu&#8217;aux concentrations comprises entre 0,02 M et 0,1 M l&#8217;eau oxyg&#233;n&#233;e r&#233;duisait tous les param&#232;tres morphom&#233;triques mesur&#233;s, &#224; savoir : la germination des graines, les croissances longitudinale et tangentielle des tiges et les surfaces foliaires des plantes. Aux faibles concentrations de 0,01 M, ce compos&#233; a toutefois provoqu&#233; une l&#233;g&#232;re et non significative stimulation des param&#232;tres &#233;tudi&#233;s. Dans les gammes de concentrations utilis&#233;es, l&#8217;eau oxyg&#233;n&#233;e n&#8217;est donc pas recommandable pour l&#8217;am&#233;lioration des productions de roselle au Gabon.Mots cl&#233;s: Roselle, eau oxyg&#233;n&#233;e, graines, tiges, feuilles, croissanc

Matrix Metalloproteinase 13 Is Induced in Fibroblasts in Polyomavirus Middle T Antigen-Driven Mammary Carcinoma without Influencing Tumor Progression

Matrix metalloproteinase (MMP) 13 (collagenase 3) is an extracellular matrix remodeling enzyme that is induced in myofibroblasts during the earliest invasive stages of human breast carcinoma, suggesting that it is involved in tumor progression. During progression of mammary carcinomas in the polyoma virus middle T oncogene mouse model (MMTV-PyMT), Mmp13 mRNA was strongly upregulated concurrently with the transition to invasive and metastatic carcinomas. As in human tumors, Mmp13 mRNA was found in myofibroblasts of invasive grade II and III carcinomas, but not in benign grade I and II mammary intraepithelial neoplasias. To determine if MMP13 plays a role in tumor progression, we crossed MMTV-PyMT mice with Mmp13 deficient mice. The absence of MMP13 did not influence tumor growth, vascularization, progression to more advanced tumor stages, or metastasis to the lungs, and the absence of MMP13 was not compensated for by expression of other MMPs or tissue inhibitor of metalloproteinases. However, an increased fraction of thin collagen fibrils was identified in MMTV-PyMT;Mmp13−/− compared to MMTV-PyMT;Mmp13+/+ tumors, showing that collagen metabolism was altered in the absence of MMP13. We conclude that the expression pattern of Mmp13 mRNA in myofibroblasts of invasive carcinomas in the MMTV-PyMT breast cancer model recapitulates the expression pattern observed in human breast cancer. Our results suggest that MMP13 is a marker of carcinoma-associated myofibroblasts of invasive carcinoma, even though it does not make a major contribution to tumor progression in the MMTV-PyMT breast cancer model

Identical temperature dependence of the time scales of several linear-response functions of two glass-forming liquids

The frequency-dependent dielectric constant, shear and adiabatic bulk moduli, longitudinal thermal expansion coefficient, and longitudinal specific heat have been measured for two van der Waals glass-forming liquids, tetramethyl-tetraphenyl-trisiloxane (DC704) and 5-polyphenyl-4-ether. Within the experimental uncertainties the loss-peak frequencies of the measured response functions have identical temperature dependence over a range of temperatures, for which the Maxwell relaxation time varies more than nine orders of magnitude. The time scales are ordered from fastest to slowest as follows: Shear modulus, adiabatic bulk modulus, dielectric constant, longitudinal thermal expansion coefficient, longitudinal specific heat. The ordering is discussed in light of the recent conjecture that van der Waals liquids are strongly correlating, i.e., approximate single-parameter liquids.Comment: 8 pages, 6 figures, Substantially revised versio