Corneal endothelial cell loss between the Kongsap manual phacofragmentation and phacoemulsification.

OBJECTIVE: To compare corneal endothelial cell loss between the Kongsap manual phacofragmentation and phacoemulsification. MATERIAL AND METHOD: One hundred two eyes with age-related cataract were randomized to undergo either the Kongsap manual phacofragmentation (Group 1, 52 eyes) or phacoemulsification surgery (Group 2, 50 eyes) with implantation of a posterior chamber, foldable, acrylic intraocular lens performed by one surgeon. The main parameters were corneal endothelial cell density (ECD), best corrected visual acuity (BCVA), and intraoperative and postoperative complications. Follow-up visits were scheduled at 1, 4, and 12 weeks. RESULTS: Pre-operatively, the mean ECD in Group 1 was 2,350 +/- 229 cells/mm2 and in Group 2 was 2,429 +/- 263 cells/mm2 (p = 0.112). Mean ECD decrease was 7.61% in Group 1 and 7.19% in Group 2 at the end of 12 weeks. The 95% confidence intervals of the mean differences of the endothelial cell loss at 4 weeks and 12 weeks after surgery were -1.87 to 2.04% and -2.77 to 3.63%, respectively. Mean best-corrected visual acuity at the end of 4 weeks was 0.88 +/- 0.22 in Group 1 and 0.82 +/- 0.24 in Group 2 (p = 0.117). There was no statistical difference between the groups in intra-operative and postoperative complications (p > 0.05). CONCLUSION: The corneal endothelial cell loss after cataract surgery with the Kongsap manual phacofragmentation is equivalent to those of phacoemulsification and both surgical techniques allowed excellent visual results.

Disfunción edoltelial inducida por hipercolesterolemia: esudio in vitroen un modelo animal / Endothelial dysfunction induced by hypercholesterolemia: in vitro study ina animal model

Los factores de riesgo tradicionales como hipercolesterolemia afectan la producción endotelial o acción de sustancias vasoactivas fundamentales como el óxido nítrico (NO). Este fenómeno tiene efectos proa terogénicos y protombóricos. El objetivo del presente estudio fue evaluar los efectos in vitro de la hipercolesterolemia sobre la función endotelial y la formación de placa ateroesclerótica en tejido aórtico proveniente de conejos hipercolesterolémicos.Los resultados encontrados sugieren que los niveles elevados de colesterol sérico producen una alteración en la vasodilatación arterial dependiente de endotelio funcional, la cual se presenta sin cambios morfológicos de las células endoteliales. Se corrobora que la hipercolesterolemia es un factor de riesgo independiente de ateroesclerosis el cual contribuye a la disfunción como evento inicial en la génesis del proceso

Dose-dependent effect of resveratrol on proliferation and apoptosis in endothelial and tumor cell cultures

Experimental data suggest that Resveratrol, a compound found in grapes and other fruits may influence cell proliferation and apoptosis. The aim of our experiments was to study the effect of Resveratrol on tumor cell cultures and an endothelial cell culture in order to examine the effect of various doses of this compound on active cell death and cell proliferation. Human tumor (HT-29, SW-620, HT-1080) and endothelial (HUV-EC-C) cells were treated with various doses of (0.1 to 100.0 microg/ml) Resveratrol in vitro. Cell number, apoptotic and mitotic index was measured 24, 48 and 72 h after treatment. Low doses (0.1-1.0 microg/ml) of Resveratrol enhance cell proliferation, higher doses (10.0-100.0 microg/ml) induce apoptosis and decrease mitotic activity, which is reflected in changes of cell number. Resveratrol influences dose dependently the proliferative and apoptotic activity of human tumor and endothelial cells. The possible role of formaldehyde in the mechanism of action of Resveratrol is discussed.

Temperature-Responsive surface for novel co-culture systems of hepatocytes with endothelial cells: 2-D patterned and double layered co-cultures

We have developed two novel cell co-culture system, without any on cell type combination limitation, utilizing a polymer surface which is temperature-sensitive with respect to its cell adhesion characteristics. One system involves a patterned co-culture of primary hepatocytes with endothelial cells utilizing patterned masked of the electron-beam cured, temperature-responsive polymer, poly (N-isopropylacrylamide) (PIPAAm) by masked electron beam irradiation. Hepatocytes were cultured to confluency at 37 degrees C on these surfaces. When the culture temperature was reduced below 32 degrees C, cells detached from the PIPAAm-grafted areas without any need for trypsin. Endothelial cells were then seeded onto the same surfaces at 37 degrees C. These subsequently seeded endothelial cells adhered only to the now-exposed PIPAAm-grafted domains and could be co-cultured with the hepatocytes initially seeded at 37 degrees C in well-ordered patterns. The other system involves a double layered co-culture obtained by overlaying endothelial cell sheets of the designed shape onto hepatocyte monolayers. The endothelial cells adhered and proliferated on the PIPAAm-grafted surface, as on polystyrene tissue culture dishes at 37 degrees C. By reducing the temperature, confluent monolayers of cells detached from the PIPAAm surfaces without trypsin. Because the recovered cells maintaed intact cell-cell junctions together with deposited extracellular matrix, the harvested endothelial cell sheets, with designed shapes, were transferable and readily adhered to hepatocyte monolayers. Stable double layered cell sheets could be co-cultivated. These two co-culture methods enabled long-term co-culture of primary hepatocytes with endothelial cells. Hepatocytes so co-cultured with endothelial cells maintained their differentiated functions, such as albumin synthesis for unexpectedly long periods. These novel two co-culture systems offer promising techniques for basic biologic researches upon intercellular communications, and for the clinical applications of tissue engineered constructs.

Estudo das vias de co-estimulaçäo envolvidas na proliferaçäo linfocitária por células endoteliais humanas, na presença de fitohemaglutinina / Study the costimulatory pathways involved in the activation of CD4+T cells human in the presence of phytohemmagglutin

A ativação de linfócitos T envolve a ligação do receptor dos linfócitos T (TCR) ao complexo CMH/peptídeo antigênico, denominado de primeiro sinal, e a interação de moléculas de superfície presente nas células endoteliais e seus ligantes linfocitários, o segundo sinal, Ou simplesmente co-sinal. Ausência do sinal induz As células endoteliaís são capazes de capturar, processar e apresentar antígenos aos linfócitos T, com capacidade de co-estimulação superior aos fibroblastos, células musculares lisas e epiteliais. Classicamente, as células endotelais ativam células T de memória e induzem anergia em células T naives. Estudamos as vias de co-estimuiação envolvidas na proliferação linfocitária induzida por células endoteliais humanas. Linfócitos T CD4+ e CD8+ de adultos e linfócitos T CD4+ e T CD4+/CD45RA+ de cordão umbilical foram utilizados como células respondedoras na cultura mista (CMEL). Células endoteliais imortalizadas, ECV-304, e as derivadas da veia do cordão umbilical, HUVEC, foram usadas COMO células estimuladoras. Linfócitos T CD4+ naives foram obtidos por seleção positiva com anticorpos anti-CD45RA e posterior depleção com anti-CD45RO. A pureza desta subPopulação foi de 98 por cento. PHA na concentração de 5OnglmL foi acrescentada na CMEL. A Proliferação foi medida por incorporação de timidina triciada. Anticorpos monoclonais foram utilizados contra as moléculas de co-estimulação presentes nas células endoteliais e linfocitárias. Foi calculada a porcentagem de inibição da Proliferação linfocitária dê cada anticorpo em comparação ao seu controle isotípico. Células endoteliais induziram Proliferação das células respondedoras somente na presença de PHA. Nas CMEL com ECV-304 e linfócitos T CD4+ de adulto, obtivemos as porcentagens de inibição com os seguintes anticorpos: anti-CD2 (clone T11; 98.2+12.4), anti-CD58 (clone BRlC5; 57.6+-11), anti-CD59 (MEM43; 50+11), anti-CD11a (AFOL-1; 70.7+15), anti-CDl8 (7E4; 58.9+16.6), anti-CD54 (BBIG-1L; 68.8+15.6), anti-CD102 (BT-1; 30.9-+11.9) and anti-CDl54 (24-31; 26.7-+11.9). Não obtivemos inibição nesta proliferação, com os AcMo dirigidos contra: CD27, CD3O, CD7O, CD28, CDl37, CDlO6, CD29, CDlO6, CD49d e com a proteína de fusão CTLA4-ig. Nas CMEL com HUVEC e linfócitos T CD4+ de sangue de cordão, obtivemos as seguintes porcentagens de inibição: anti-CD2 (clone T11, 64.8+16.1), anti-CD58 (clone TS2/9; 63.6ñ13.6), anti-CD59 (MEM43; 36ñ10.9), anti-Cd11a (AFOL-1; 33.4ñ12, anti-CD18 (7E4; 25ñ11.4...(au)

The induction of cyclooxygenase-2 (COX-2) in cultured endothelial cells treated with serum from preeclampsia is mediated by interleukin-6.

COX-2 protein, but not COX-1 protein, was induced in HUVEC from women with a normal pregnancy (nHUVEC) treated with serum from patients with preeclampsia (pSerum), but not with serum from women with a normal pregnancy (nSerum). COX activity in pSerum treated nHUVEC was less than in nSerum treated nHUVEC. Interestingly, the induction of COX-2 protein in nHUVEC treated with pSerum was inhibited by antiIL-6 antibody. The decreased COX activity in nHUVEC treated with pSerum plus antiIL-6 antibody was also reversed in a dose dependent manner. Thus, the induction of COX-2 in pSerum treated nHUVEC was mediated by IL-6. Therefore, the development of selective inhibitors of COX-2 or of IL-6 antagonists may have a potential role in the prevention and treatment of preeclampsia.

Ultrastructural studies of mast cells, fibroblasts and endothelial cells, in clinically involved skin of patients with scleroderma

A hypothetical role of mast cells, endothelial cells and fibroblasts, in the pathophysiology of scleroderma has been proposed. The present study aimed at the illustration of the ultrastructural changes of these cells, in particular, in the skin of patients with scleroderma. In the following work, electron microscopic examination demonstrated a spectrum of degranulated mast cells as well as different grades of cell-activation evidenced by presence of hypodense granules with increased cytoplasmic organelles such as rough-surface endoplasmic reticulum [RER], polysome and mitochondria. The fibroblasts showed numerous dilated RER, intra and extracytoplasmic filaments. Some fibroblast - like cells were seen ingesting some mast cell granules. Evidences of endothelial injury were detected in the form of thickening and stratification of basal lamina and swollen endothelial cells

Decreased biological activity is related to abnormal multimeric structure of von Willebrand factor in pulmonary hypertension

We evaluated the correlation between decreased biological activity and abnormalities in the multimeric structure of plasma von Willebrand factor (vWF) in 27 pulmonary hypertensive patients (median age, 21 years). The biological activity of vWF was measured by the ristocetin cofactor assay and its multimeric structure was assessed by Western immunoblotting after SDS-agarose gel electrophoresis. In spite of high antigenic activity of vWF in plasma (139 ñ 65 vs 91 ñ 27 per cent in controls, P=0.003), the biological activity expressed as a percent of the control value was decreased in pulmonary hypertensive patients (60-88 per cent activity, 95 per cent CI for the mean). High molecular weight multimers (biologically active forms) were absent in patients and there was a significant increase in the concentration of low molecular weight polymers in comparison with normals (56 ñ 12 and 35 ñ 12 per cent of total multimer density, respectively, P<0.001). Multimeric abnormalities were positively correlated with plasma vWF levels (r=0.51, P=0.0007) and negatively correlated with vWF biological activity (r=-0.54, P=0.004). Thus, decreased biological function is related to abnormalities in the multimeric structure of vWF, possibility reflecting extensive endothelial dysfunction in pulmonary hypertension.

Some cytological answers to atherosclerosis

A concise review of the atherosclerotic process is presented and a new concept is proposed as to the fundamental nature of atheromas and the process of atheromatosis. Evidence and arguments are provided to consider the atheroma as a newly developing and evolving vasculo-endocrine unit of the vessel wall, secreting steroidal chemicals and hormones into the blood stream. These hormones are of probable importance as trophic factors for the vascular tunica interna and are also of circulatory value for enhancing trans-capillary transport of nutrients and metabolites and catalysing the tissue reactions of metabolism in the tissue spaces [thus being tissue circulation promoters and enhancers]. They may be of use in preventing or treating various tissue metabolic storage disorders as well as in creating better general and special tissues for the body

Three possible triggers to induce thrombocytopenia in dengue virus infection.

Changes in platelet count by dengue virus-platelet interaction and the participation of anti-DV antibody to such changes were studied in vitro with the aim to investigate a mechanism of how the DV causes prominent thrombocytopenia characteristically seen in DHF. The results obtained showed that: (1) DV antigen attached to human platelets without immune-mediated reaction, (2) a decrease in platelet count was more markedly demonstrated by the binding of anti-DV antibody on the DV antigen associated with platelets than by the binding of the antigen-antibody complex on platelets, (3) a modulation of endothelial cell by the infection of DV to the cell was suggested as one of the causes of the thrombocytopenia.

Fluorescent low density lipoprotein and acetylated low density lipoprotein labeling of cultured bovine trabecular endothelial cells

A preliminary study was performed to investigate the staining characteristics of trabecula. endothelial cells with low density lipoprotein (LDL) and acetylated low density lipoprotein (Ac-LDL) labeled with a fluorescent probe, 1, 1`- dioctadecyl-3,3,3`, 3`- tetramethyl-indocarbocyanine perchlorate (Dil). Trabecular endothelial cells revealed a strong fluorescence with Dil-LDL, which was contradictory to the previous results obtained from other types of endothelial cells. These cells also showed a moderate fluorescence with Dil-Ac-LDL. Scleral fibroblasts and keratocytes showed a moderate to strong fluorescence with Dil-LDL and a weak fluorescence with Dil-Ac-LDL. Corneal endothelial cells revealed a very weak background fluorescence with Dil-LDL and a moderate fluorescence with Dil-Ar-LDL. Therefore, these four kinds of cells could not be definitely differentiated depending only on the staining characteristics with Dil-LDL and Dil-Ac-LDt.