Expression of Integrin and TGFBI in Human Retinal Pericytes

Purpose: The aim for this study is to investigate the expression of integrin α3, β1 and TGF-β induced protein (TGFBI) and the secretion of TGFBI by primary culture of human retinal pericytes (pHRP). Evidence suggests that chronic diabetes associate with HRP apoptosis leading to the development of diabetic retinopathy. Methods: pHRP (Cell Systems) were cultured in complete media (15mM glucose) in a humidified, 5% CO2, 37°C condition. Cells were seeded at passage 6 to 8 into a 24 well-plate with coverslips or P10 dishes. Cells (85% confluence) media were then replaced by DMEM media with euglycemic glucose (5.5mM) or hyperglycemic glucose (30mM) and cells were incubated for 48 or 72 hours. Gene and protein expressions of α3, β1 were detected by Real-Time PCR and flow cytometry. TGFBI gene expression was detected by Real-Time PCR and ELISA was used to measure protein level in cell media. Results: Real-Time PCR showed expression of α3, β1 and TGFBI in pHRP at 48 hrs of incubation in both glucose concentrations. Expression of a3 in pHRP in 30 mM glucose was 1.3 times higher than cells in 5.5mM glucose whereas expressions of b1 and TGFBI were comparable in two glucose concentrations. Flow cytometry results also showed expression of integrin subunits in pHRP at 72 hr of incubation. Expression of a3 in pHRP in 30mM glucose was similar to those in cells in 5.5m M (MFI of 251 vs 221 respectively). However, expression of b1 was higher in cells in the higher glucose concentration (MFI: 422 vs 343). ELISA data showed secretion TGFBI protein by HRP at 48 hr of incubation. Protein concentration in media of cell in 30mM glucose was significantly higher than those in 5.5mM (97 vs 57 pg/ml; p=0.0318). Conclusions: This is the first report on the expression of integrin subunits in HPR in euglycemic and hyperglycemic conditions. Both RT-PCR and flow cytometry results show α3, β1 subunits expressions, the level of which may be affected by glucose concentration in the cell media. Furthermore, our ELISA results confirm the secretion of TGFB1 by HRP and a significantly higher protein secretion in hyperglycemic condition. Overall, our data support the hypothesis of integrin and TGFBI expression in HRP. The increase in TGFBI secretion in hyperglycemia suggest a possible role of diabetes. Further studies will provide insight into the role of integrin and TGFBI interaction on the signaling pathway of HRP apoptosis and diabetic retinopathy

Grout long radius flow testing to support Saltstone disposal Unit 5 design

The Saltstone Facility, located within the Savannah River Site (SRS) near Aiken, South Carolina, consists of two facility segments: The Saltstone Production Facility (SPF) and the Saltstone Disposal Facility (SDF). The SPF receives decontaminated legacy low level sodium salt waste solution that is a byproduct of prior nuclear material processing. The salt solution is mixed with cementitious materials to form a grout slurry known as “Saltstone”. The grout is pumped to the SDF where it is placed in a Saltstone Disposal Unit (SDU) to solidify. SDU 6 is referred to as a “mega vault” and is currently in the design stage. The conceptual design for SDU 6 is a single cell, cylindrical geometry approximately 114.3 meters in diameter by 13.1 meter high and is larger than previous cylindrical SDU designs, 45.7 meters in diameter by 7.01 meters high (30 million gallons versus 2.9 million gallons of capacity). Saltstone slurry will be pumped into the new waste disposal unit through roof openings at a projected flow rate of about 34.1 cubic meters per hour. Nine roof openings are included in the design to discharge material into the SDU with an estimated grout pour radius of 22.9 to 24.4 meters and initial drop height of 13.1 meters. The conceptual design for the new SDU does not include partitions to limit the pour radius of the grout slurry during placement other than introducing material from different pour points. This paper addresses two technical issues associated with the larger diameter of SDU 6; saltstone flow distance in a tank 114.3 meters in diameter and quality of the grout. A long-radius flow test scaled to match the velocity of an advancing grout front was designed to address these technology gaps. The emphasis of the test was to quantify the flow distance and to collect samples to evaluate cured properties including compressive strength, porosity, density, and saturated hydraulic conductivity. Two clean cap surrogate mixes (saltstone premix plus water) were designed to simulate slurry with the reference saltstone rheology and a saltstone with extra water from the process flushing operation. Long-radius flow tests were run using approximately 4.6 cubic meters of each of these mixes. In both tests the pump rate was 0.063 liters/second (1 gpm). A higher pump rate, 0.19 liters/second (3 gpm), was used in a third long-radius flow test. The angle of repose of the grout wedges increased as a function of time in all three tests. The final angles of repose were measured at 3.0º, 2.4º, and 0.72º. The pump rate had the largest effect on the radial flow distance and slope of the grout surface. The slope on the pour placed at 0.19 liters/second (3 gpm) was most representative of the slope on the grout currently being pumped into SDU 2 which is estimated to be 0.7º to 0.9º. The final grout heights at 1/3 of a meter from the discharge point were 115, 105, and 38 cm. Entrapped air (≥ 0.25 cm bubbles) was also observed in all of the mixes. The entrapped air appeared to be released from the flows within about 3.1 meters (10 feet) of the discharge point. The bleed water was clear but had a thin layer of floating particulates. The bleed water should be retrievable by a drain water collection system in SDU 6 assuming the system does not get clogged. Layering was observed and was attributed to intervals when the hopper was being cleaned. Heat from the hydration reactions was noticeable to the touch

Hemorragia de vías digestivas altas de urgencia en el HUN: correlación clínico-endoscópica

Acute bleeding of upper digestive tract (ABUDT) is one of the most frequent emergencies at emergency services. Determining what patients have bleeding is an important challenge for clinicians since this it may be a risk for the patient´s life. Bleeding requires an interdisciplinary approach including medical, endoscopic and surgical management.  This study shows the relevance of carrying out upper digestive endoscopies of emergency, its endoscopic findings and the need of therapeutic management. Reports of 189 endoscopies diagnosed as having ABUDT were evaluated, from January 1 to June 30, 2010 at the digestive endoscopy unit of Hernando Moncaleano Perdomo University Hospital.  There were patients aged from 1 to 93, predominantly male (53%). Patients having no urgent bleeding cause was of 60,4%, urgent cause was of 39,6% from which 29.1% had non-variceal bleeding and 10.5% had variceal bleeding. Endoscopic therapeutics was not performed on patients having non-variceal bleeding; on the contrary, endoscopic therapeutics was performed on 90% of patients having variceal bleeding. Gastritis (26%) was the most frequent diagnosed cause of bleeding, following gastric ulcer (12%). Patients admitted to emergency service having ABUDT should be evaluated in an integral form using protocols to determine high-risk patients that require to have an upper emergency digestive endoscopy.La hemorragia aguda de vías digestivas altas (HVDA) es una de las emergencias más frecuentes en los servicios de urgencias(1). Un reto importante para el clínico es evaluar que pacientes presentan un episodio hemorrágico que puede poner en riesgo la vida del paciente y sobre el que se debe actuar en forma interdisciplinaria con manejo médico, endoscópico y quirúrgico(2,3). El presente estudio muestra la pertinencia de la realización de endoscopias digestivas altas de urgencia, sus hallazgos endoscópicos y necesidad de manejo terapéutico. Se evaluaron los informes de 189 endoscopias realizadas con diagnóstico de HVDA de enero a junio de 2010 en la unidad de endoscopia digestiva del Hospital Universitario Hernando Moncaleano Perdomo (HUN). Se encontraron pacientes entre los 1 y 93 años, con predominio del género masculino (53%). Encontramos al 60,4% de los pacientes sin causa urgente de sangrado, el 39,6% presentó causa urgente, de los cuales 29,1% con sangrado no variceal, y 10,5% con sangrado variceal, se realizó terapéutica endoscópica a diez pacientes (5,1%) con sangrado no variceal, con sangrado variceal se realizó terapéutica endoscópica a 17 (9%). El diagnóstico más frecuente de las causas no urgentes de sangrado fue gastritis con un 26%, seguido de ulcera gástrica con un 12%. Los pacientes que ingresan al servicio de urgencias con diagnóstico de HVDA deben ser evaluados en forma íntegra con creación de protocolos para establecer cuales pacientes son de alto riesgo y ameriten la realización de una endoscopia digestiva alta de urgencia

Dynamic and Thermodynamic Influences on Precipitation in Northeast Mexico on Orbital to Millennial Timescales

The timing and mechanisms of past hydroclimate change in northeast Mexico are poorly constrained, limiting our ability to evaluate climate model performance. To address this, we present a multiproxy speleothem record of past hydroclimate variability spanning 62.5 to 5.1 ka from Tamaulipas, Mexico. Here we show a strong influence of Atlantic and Pacific sea surface temperatures on orbital and millennial scale precipitation changes in the region. Multiple proxies show no clear response to insolation forcing, but strong evidence for dry conditions during Heinrich Stadials. While these trends are consistent with other records from across Mesoamerica and the Caribbean, the relative importance of thermodynamic and dynamic controls in driving this response is debated. An isotope-enabled climate model shows that cool Atlantic SSTs and stronger easterlies drive a strong inter-basin sea surface temperature gradient and a southward shift in moisture convergence, causing drying in this region

Multiple strategies to prevent oxidative stress in Arabidopsis plants lacking the malate valve enzyme NADP-malate dehydrogenase

The nuclear-encoded chloroplast NADP-dependent malate dehydrogenase (NADP-MDH) is a key enzyme controlling the malate valve, to allow the indirect export of reducing equivalents. Arabidopsis thaliana (L.) Heynh. T-DNA insertion mutants of NADP-MDH were used to assess the role of the light-activated NADP-MDH in a typical C3 plant. Surprisingly, even when exposed to high-light conditions in short days, nadp-mdh knockout mutants were phenotypically indistinguishable from the wild type. The photosynthetic performance and typical antioxidative systems, such as the Beck–Halliwell–Asada pathway, were barely affected in the mutants in response to high-light treatment. The reactive oxygen species levels remained low, indicating the apparent absence of oxidative stress, in the mutants. Further analysis revealed a novel combination of compensatory mechanisms in order to maintain redox homeostasis in the nadp-mdh plants under high-light conditions, particularly an increase in the NTRC/2-Cys peroxiredoxin (Prx) system in chloroplasts. There were indications of adjustments in extra-chloroplastic components of photorespiration and proline levels, which all could dissipate excess reducing equivalents, sustain photosynthesis, and prevent photoinhibition in nadp-mdh knockout plants. Such metabolic flexibility suggests that the malate valve acts in concert with other NADPH-consuming reactions to maintain a balanced redox state during photosynthesis under high-light stress in wild-type plants

Impaired response of blood neutrophils to cell-death stimulus differentiates AQP4-IgG-seropositive NMOSD from MOGAD

BACKGROUND: In neuromyelitis optica spectrum disorders (NMOSD) and myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD), neutrophils are found in CNS lesions. We previously demonstrated that NMOSD neutrophils show functional deficiencies. Thus, we hypothesized that neutrophil accumulation in the CNS may be facilitated by impairments affecting mechanisms of neutrophil death. OBJECTIVE: To evaluate cell death in blood neutrophils from aquaporin-4 (AQP4)-IgG-seropositive NMOSD and MOGAD patients as well as matched healthy controls (HC) using in vitro assays. METHODS: Twenty-eight AQP4 + NMOSD and 19 MOGAD patients in stable disease phase as well as 45 age- and sex-matched HC were prospectively recruited. To induce cell death, isolated neutrophils were cultured with/without phorbol 12-myristate 13-acetate (PMA). Spontaneous and PMA-induced NETosis and apoptosis were analyzed using 7-AAD and annexin-V by flow cytometry. Caspase-3 was assessed by western blot. Myeloperoxidase-DNA complexes (MPO-DNA), MPO and elastase were evaluated by ELISA, and cell-free DNA (cfDNA) by a fluorescence-based assay. Reactive oxygen species (ROS) were evaluated by a dihydrorhodamine 123-based cytometric assay. Serum GM-CSF, IL-6, IL-8, IL-15, TNF-ɑ and IL-10 were evaluated by multiplex assays, and neurofilament light chain (NfL) by single-molecule array assay. RESULTS: In response to PMA, neutrophils from AQP4 + NMOSD but not from MOGAD patients showed an increased survival, and subsequent reduced cell death (29.6% annexin V(+) 7-AAD(+)) when compared to HC (44.7%, p = 0.0006). However, AQP4 + NMOSD also showed a mild increase in annexin V(+) 7-AAD(-) early apoptotic neutrophils (24.5%) compared to HC (20.8%, p = 0.048). PMA-induced reduction of caspase-3 activation was more pronounced in HC (p = 0.020) than in AQP4 + NMOSD neutrophils (p = 0.052). No differences were observed in neutrophil-derived MPO-DNA or serum levels of MPO, elastase, IL-6, IL-8 and TNF-ɑ. IL-15 levels were increased in both groups of patients. In AQP4 + NMOSD, an increase in cfDNA, GM-CSF and IL-10 was found in serum. A positive correlation among cfDNA and NfL was found in AQP4 + NMOSD. CONCLUSIONS: AQP4 + NMOSD neutrophils showed an increased survival capacity in response to PMA when compared to matched HC neutrophils. Although the data indicate that the apoptotic but not the NETotic response is altered in these neutrophils, additional evaluations are required to validate this observation

Global Diversity of Brittle Stars (Echinodermata: Ophiuroidea)

This review presents a comprehensive overview of the current status regarding the global diversity of the echinoderm class Ophiuroidea, focussing on taxonomy and distribution patterns, with brief introduction to their anatomy, biology, phylogeny, and palaeontological history. A glossary of terms is provided. Species names and taxonomic decisions have been extracted from the literature and compiled in The World Ophiuroidea Database, part of the World Register of Marine Species (WoRMS). Ophiuroidea, with 2064 known species, are the largest class of Echinodermata. A table presents 16 families with numbers of genera and species. The largest are Amphiuridae (467), Ophiuridae (344 species) and Ophiacanthidae (319 species). A biogeographic analysis for all world oceans and all accepted species was performed, based on published distribution records. Approximately similar numbers of species were recorded from the shelf (n = 1313) and bathyal depth strata (1297). The Indo-Pacific region had the highest species richness overall (825 species) and at all depths. Adjacent regions were also relatively species rich, including the North Pacific (398), South Pacific (355) and Indian (316) due to the presence of many Indo-Pacific species that partially extended into these regions. A secondary region of enhanced species richness was found in the West Atlantic (335). Regions of relatively low species richness include the Arctic (73 species), East Atlantic (118), South America (124) and Antarctic (126)

Marine Biodiversity in the Caribbean: Regional Estimates and Distribution Patterns

This paper provides an analysis of the distribution patterns of marine biodiversity and summarizes the major activities of the Census of Marine Life program in the Caribbean region. The coastal Caribbean region is a large marine ecosystem (LME) characterized by coral reefs, mangroves, and seagrasses, but including other environments, such as sandy beaches and rocky shores. These tropical ecosystems incorporate a high diversity of associated flora and fauna, and the nations that border the Caribbean collectively encompass a major global marine biodiversity hot spot. We analyze the state of knowledge of marine biodiversity based on the geographic distribution of georeferenced species records and regional taxonomic lists. A total of 12,046 marine species are reported in this paper for the Caribbean region. These include representatives from 31 animal phyla, two plant phyla, one group of Chromista, and three groups of Protoctista. Sampling effort has been greatest in shallow, nearshore waters, where there is relatively good coverage of species records; offshore and deep environments have been less studied. Additionally, we found that the currently accepted classification of marine ecoregions of the Caribbean did not apply for the benthic distributions of five relatively well known taxonomic groups. Coastal species richness tends to concentrate along the Antillean arc (Cuba to the southernmost Antilles) and the northern coast of South America (Venezuela – Colombia), while no pattern can be observed in the deep sea with the available data. Several factors make it impossible to determine the extent to which these distribution patterns accurately reflect the true situation for marine biodiversity in general: (1) highly localized concentrations of collecting effort and a lack of collecting in many areas and ecosystems, (2) high variability among collecting methods, (3) limited taxonomic expertise for many groups, and (4) differing levels of activity in the study of different taxa