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[Studies on serum protein fractions of patients with maxillary sinus cancer undergoing a combination of radiotherapy and chemotherapy: II. Relationship between changes in serum protein fractions and prognosis].

We examined the correlations between changes in serum protein fractions and the prognosis of the patients. The levels of 21 protein components of the sera of 36 patients with maxillary sinus cancer were determined by a single radial immunodiffusion method before and after radiation therapy. The patients with maxillary sinus cancer were treated with combined intra-arterial infusion of bleomycin and external irradiation of 60 Co gamma-rays, and were concurrently treated with 5-fluorouracil at 200 mg/day p.o. The levels of the same protein components were also measured in 34 normal adult as a control. All patients were observed 5 years and 12 years after radiation therapy. In patients who had survived at least 5 years after radiation therapy, the Alb, Tf, Hx, IgG and IgM levels measured before radiation therapy were elevated significantly compared with those who had died within 5 years. In those who had survived at least 5 years, the Alb, Tf, Hx, IgG, IgM, IgA and I alpha I levels measured after radiation therapy were elevated significantly compared with those who had died within 5 years, and AT III was reduced. In cases of maxillary sinus cancer following a period of 5 to 12 years after radiation therapy, multiple regression analysis was used to determine whether increased concentrations of serum protein fractions were associated with good prognosis for the original disease, alpha 2HS. IgM, HX, alpha 1AT and alpha 1X before radiation therapy were positively correlated with survival, whereas AT III, Pmg, Cp, IgA, and alpha 1AG showed negative correlations. After radiation therapy, Pmg, Hx, Cp, Cl inh and Fib were found to be positive factors of survival rate, whereas alpha 2M, alpha 2Pl, I alpha 1, IgA, alpha 1AG and C3 were negative factors.

Adult↗

Plasma renin activity levels influence the effect of converting enzyme inhibitors in preserving renal function in hypertension.

OBJECTIVE: To study the effect of treatment with cilazapril versus nifedipine on the glomerular filtration rate (GFR), microalbumin excretion rate (mu Alb) and baseline plasma renin activity (PRA) levels in non-diabetic or diabetic essential hypertension. DESIGN AND METHODS: Thirty-four moderate essential hypertensives and thirty-two type II diabetic hypertensives with currently normal renal function completed the study, receiving treatment with a converting enzyme inhibitor (CEI, 5-10 mg/day cilazapril) or a non-CEI (20-40 mg slow-release nifedipine three times a day) during a 12-month controlled follow-up study. RESULTS: Either treatment induced a similar reduction in blood pressure. Patients with PRA > 3 ng/ml per h treated with cilazapril showed a proportional increase in GFR, whereas in those treated with a non-CEI there was a decline in GFR after 8 and 12 months. Only diabetic patients treated with cilazapril showed a mu Alb reduction associated with a stable GFR. CONCLUSION: PRA predicts the effect of CEI preserving renal function in essential and diabetic hypertension.

Adolescent↗

[Visual analysis of hepatic function by a scoring system using 99mTc-GSA].

To establish a simple hepatic function test, a five-point scoring system using 99mTc-galactosyl-human serum albumin (GSA) was applied to seventy-six patients with or suspected with hepatic dysfunction. Dynamic data at rest were obtained immediately and 30 minutes after intravenous bolus injection of 99mTc-GSA (185 MBq). Regions of interest (ROIs) were superimposed over the images of the liver and the entire heart. A visual analysis using a image taken 5 to 6 minutes after radiotracer injection was performed by the three observers blinded to the clinical information to assess their agreement or disagreement on the grading score. kappa-Test was used to evaluate the goodness of agreement. Three pairs in the first reading showed excellent interobserver agreement (0.92 mean kappa-value). Three pairs in the first and second reading also showed excellent intraobserver agreement (0.89 mean kappa-value). Data were also analyzed for correlations with heart/liver count ratio, ICG R15, LHL15, Alb, TB, PT, HPT and ZTT. There were good correlations between visual grading score and ICG R15 (Rs = 0.619, p < 0.0001) and between grading score and heart/liver count ratio (Rs = 0.866, p < 0.0001). Grading score was also correlated with LHL15, Alb, TB, PT, HPT and ZTT (Rs = -0.770; -0.580; 0.593; -0.544; -0.622; 0.518, p < 0.0001 for each). There was a significant difference between heart/liver count ratio and LHL15 in the Grade 4 and Grade 5 groups. In conclusion, the present data suggest that a five-point scoring system based on visual analysis using 99mTc-GSA uptake is a useful indicator for hepatic dysfunction in a clinical setting.

Aged↗

[Echographic parameters in the evaluation of the degree of portal hypertension].

We did a prospective study in thirty one patients with ascites who were hospitalized at Cayetano Heredia Hospital (H.N.C.H.). We investigated the association and correlation between ultrasonographic parameters of portal hypertension (HTP) and the presence and level of HTP (determinate by the serum-to-ascites albumin concentration gradient). In our study, we demonstrated that ultrasonographic splenomegaly studied by longitudinal diameter of the spleen discriminate patients with HTP with a high positive predictive value (94.4%), although it didn't happen with transverse diameter of the spleen. However patients with elevated measurements of high GRAD-Alb (> or = 2.35 +/- 0.05 g/dl) had alterations in the transversal diameter of the spleen with a high negative predictive value (91.7%), these findings were suggested its use like a parameter for reject high levels of HTP. In our study, we demonstrated that the existence of changes at diameters of portal vein and splenic vein could not discriminate patients with HTP. By the way, we didn't find association between degree of high GRAD-Alb and the different splenic measurements.

Adult↗

Effects of time of storage, albumin, and osmolality changes on outflow facility (C) of bovine anterior segment in vitro.

PURPOSE: To analyze the influence of time of storage, the presence of albumin at physiological concentrations, and the perfusion with anisosmotic media on the aqueous humor outflow facility (C) of isolated bovine anterior segments (AS). METHODS: Anterior segments dissected from cow eyes were perfused at a constant pressure of 10 mm Hg with Dulbecco's modified Eagle's medium (DMEM; osmolality 300 mOsm/kg), with hyposmotic media (150, 210, and 270 mOsm/kg), or with hyperosmotic media (360, 420, and 480 mOsm/kg). Outflow facility was calculated every 5 seconds as the ratio between average inflow from the reservoir (in microliters per minute) and the perfusion pressure (in millimeters of mercury). Three groups were studied: a 0-hour group, with AS perfused with DMEM 1 to 3 hours after enucleation; a 0-hour alb-group, with AS perfused with DMEM plus 0.1 mg/ml albumin 1 to 3 hours after enucleation; and a 24-hour group, with AS perfused after storage for 24 hours in DMEM. In the 0-hour groups, perfusion with increasingly hyposmotic or hyperosmotic media was also made in 30-minute steps, followed by a return to isosmotic medium for 90 minutes. RESULTS: Perfusion of AS with DMEM for 9 hours caused a progressive increase in C that was statistically significant at 225 minutes in the 0-hour group perfused with DMEM and at 195 minutes in the 24-hour group perfused with DMEM. The 0-hour alb-group perfused with DMEM did not show changes in C throughout the 9-hour perfusion period. Perfusion with increasingly hyposmotic media induced a progressive decrease in C that did not recover on return to isotonic medium. Hyperosmotic media caused a progressive increase in C that returned to control values when isotonic medium was again perfused. CONCLUSIONS: Preservation of tissue for C measurements is best achieved with short storage time (1 to 3 hours). Physiological concentrations of albumin (0.1 mg/ml) prevent development of washout, suggesting that albumin or an albumin-bound factor in aqueous humor may play a role in the maintenance of outflow resistance. Outflow facility also may be influenced by volume changes in the trabecular meshwork.

Animals↗

Assessment of monoethylglycinexylidide as measure of liver function for patients with chronic viral hepatitis.

The liver metabolizes lidocaine by oxidative deethylation to form monoethylglycinexylidide (MEGX), an analyte proposed as an index of liver function. We determined MEGX and lidocaine serum concentrations with the TDx (Abbott Laboratories) at baseline and 15, 30, 60, and 90 min after the intravenous administration of lidocaine (1 mg/kg), analyzing specimens from 12 apparently healthy volunteers and 40 patients with chronic viral hepatitis diagnosed by liver biopsy and serum tests. The patients were grouped on the basis of the histology activity index. The following laboratory tests were performed on serum specimens from all subjects: albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase, total bilirubin, and prothrombin time. The results showed no significant difference among the four groups for the concentrations of MEGX, lidocaine, and lidocaine/MEGX at the four time points. However, the concentrations of ALB, ALT, AST, AST/ALT, and prothrombin time were substantially different among the four groups. Thus, we conclude that assay of MEGX in our patients with chronic viral hepatitis did not contribute to the assessment of liver function when compared with apparently healthy volunteers and traditional tests of liver function.

Adult↗

[The protein-synthesizing activity of a number of strains of Cladosporium cladosporioides (Fres.) de Vries grown under conditions of continuous low-intensity gamma irradiation].

Phenomena of melanization of radioactively polluted soils, due to prevalence of melanin-containing species and positive radiotropism of some micromycetes have been found during monitoring of mycobiota of the 30-km alienation zone of the Chernobyl NPP for 10 years. To elucidate the contribution of the melanin system to the cell protection against irradiation, the influence of gamma-irradiation on the activity of protein synthesis in four Cladosporium cladosporioides (Fres.) de Vries strains has been investigated. Two strains isolated from radioactively polluted substrates were characterised by the presence of positive radiotropism. Laboratory strain 396 and its alb-mutant (with blocked melanin synthesis), did not possess this feature. The protein synthesizing activity was assayed by incorporation of 14C-leucine in the protein fraction of mycelium, grown during 7 days under continuous gamma-irradiation of low intensity and without it. The protein synthesis was activated in the radioactively treated mycelium of dark-pigmented C. cladosporioides strains and it was suppressed in similarly treated mycelium of alb-mutant of C. cladosporioides. The rate of 14C-leucine incorporation into biomass of investigated strains correlated with positive radiotropism. The dependence between protein synthesis intensity and the availability of melanin protection system in micromycetes is assumed.

Carbon Radioisotopes↗

Carbohydrate metabolism in Dictyostelium discoideum: II. Systems' analysis.

A transition model of carbohydrate metabolism during differentiation of Dictyostelium discoideum was described in Wright & Albe (1994, J. theor. Biol. 168, 231-241). From this base model, two steady-state models are derived in which concentration and flux are consistent with culmination (600 min) or young sorocarp (700 min) stages of development. These steady-state models are analyzed using metabolic control theory (MCT), and the summation property is met for metabolites, individual flux rates, and overall flux control coefficients. The MCT analyses show that flux control differs between the two stages. At culmination, cellulose synthetase is the major control point for overall pathway flux (C = 0.86) whereas at young sorocarp, glycogen phosphorylase is the major control point (C = 0.89). This is a reflection of the increased competition between the endproduct synthases, which are used to define overall pathway flux. Thus the specific metabolic state of the cells is important in assessing control distribution. The significance of this work in understanding the mechanisms controlling biochemical differentiation is discussed and related to the work of others in the field. For example, mutant analysis can reveal which enzymes are essential to differentiation (most are) but cannot reveal which enzymes are unique and/or rate-controlling.

Animals↗

Catalytic asymmetric synthesis of 19,20-dihydroakuammicine.

An enantiocontrolled total synthesis of 19,20-dihydroakuammicine using a catalytic asymmetric Michael addition of dimethyl malonate to cyclohexenone as the key step is described. The above catalytic asymmetric Michael addition proceeds quite efficiently in the presence of a heterobimetalic asymmetric catalyst (ALB-KO-t-Bu-MS 4A, 0.3 mol%), giving the corresponding Michael adduct in 94% yield and 99% ee.

Journal Article↗

Effect of ultrasound-induced hyperthermia and cis-diamminedichloride platinum II on murine renal function.

Murine renal function was evaluated after ultrasound-induced kidney hyperthermia (42.5 degrees C and 46.5 degrees C for 35 minutes) and the administration of cis-diamminedichloride platinum II (8 mg/kg). A quantitative immunonephelometric technique was employed to determine urinary total protein (TP) and albumin (Alb) 1-180 days post-treatment. Hyperthermia of 46.5 degrees C elevated urinary TP excretion significantly more than that of 42.5 degrees C cis-diamminedichloride platinum (Cis DDP) administration greatly increase urine TP with a peak mean TP concentration of 488 microgram/ml, four days after Cis DDP (normal range was 26-48 microgram/ml). This returned to normal by day 14. The TP excretion after 42.5 degrees C hyperthermia with concurrent Cis DDP was similar to Cis DDP alone, implying there was no potentiation of early or late Cis-DDP-induced renal damage by hyperthermia. Late mouse mortality was greater (P = 0.08) after Cis DDP alone, than Cis DDP with 42.5 degrees C hyperthermia. There was a statistically significant increase (P less than 0.05) in the Alb/TP excretion ratio after the addition of 42.5 degrees C hyperthermia to Cis DDP, implying an alteration in the site or nature of the Cis DDP renal lesion by hyperthermia.

Albuminuria↗

The genetic demography of the Gainj of Papua New Guinea. I. Local differentiation of blood group, red cell enzyme, and serum protein allele frequencies.

Allele frequencies are reported for 19 blood group, red cell enzyme, and serum protein loci (ABO, Rh, MN, Hb-A, LDH-A, LDH-B, SOD, PGM-1, PGM-2, 6PGD, GPT, ESD, ADA, ACP, PGK, MDH, Alb, Hp, and Tf) determined from 310 blood samples collected among the Gainj, a small population of tribal horticulturalists from highland Papua New Guniea. Fourteen of these loci display genetic variants, and ten of them are sufficiently polymorphic to permit a preliminary analysis of Gainj population structure. Patterns of variation among subdivisions of the population are analyzed using an approach analogous to a multivariate analysis of variance with unbalanced design, and weighted genetic distances are extracted from the results. The distance analysis indicates that patterns of genetic variation within this population reflect the geographical distribution of subdivisions, as well as subdivision size and movement among subdivisions. A parallel analysis of the Gainj and two other tribal groups from highland New Guinea, the Murapin Enga and the Simbai Valley Maring, suggests that the Gainj are both genetically divergent from neighboring populations and internally highly differentiated.

Adult↗

Vascular gene expression and phenotypic correlation during differentiation of human embryonic stem cells.

The study of the cascade of events of induction and sequential gene activation that takes place during human embryonic development is hindered by the unavailability of postimplantation embryos at different stages of development. Spontaneous differentiation of human embryonic stem cells (hESCs) can occur by means of the formation of embryoid bodies (EBs), which resemble certain aspects of early embryos to some extent. Embryonic vascular formation, vasculogenesis, is a sequential process that involves complex regulatory cascades. In this study, changes of gene expression along the development of human EBs for 4 weeks were studied by large-scale gene screening. Two main clusters were identified-one of down-regulated genes such as POU5, NANOG, TDGF1/Cripto (TDGF, teratocarcinoma-derived growth factor-1), LIN28, CD24, TERF1 (telomeric repeat binding factor-1), LEFTB (left-right determination, factor B), and a second of up-regulated genes such as TWIST, WNT5A, WT1, AFP, ALB, NCAM1. Focusing on the vascular system development, genes known to be involved in vasculogenesis and angiogenesis were explored. Up-regulated genes include vasculogenic growth factors such as VEGFA, VEGFC, FIGF (VEGFD), ANG1, ANG2, TGFbeta3, and PDGFB, as well as the related receptors FLT1, FLT4, PDGFRB, TGFbetaR2, and TGFbetaR3, other markers such as CD34, VCAM1, PECAM1, VE-CAD, and transcription factors TAL1, GATA2, and GATA3. The reproducibility of the array data was verified independently and illustrated that many genes known to be involved in vascular development are activated during the differentiation of hESCs in culture. Hence, the analysis of the vascular system can be extended to other differentiation pathways, allocating human EBs as an in vitro model to study early human development.

Antigens, CD34↗

Conditional knockout of the mouse NADPH-cytochrome p450 reductase gene.

NADPH-cytochrome P450 reductase (CPR or POR) is the obligatory electron donor for all microsomal cytochrome P450 (CYP or P450)-catalyzed monooxygenase reactions. Disruption of the mouse Cpr gene has been reported to cause prenatal developmental defects and embryonic lethality. In this study, we generated a mouse model with a floxed Cpr allele (termed Cpr(lox)). Homozygous Cpr(lox) mice are fertile and without any histological abnormality or any change in CPR expression. The floxed Cpr allele was subsequently deleted efficiently by crossing Cpr(lox) mice with transgenic mice having liver-specific Cre expression (Alb-Cre); the result was a decrease in the level of CPR protein in liver microsomes. The Cpr(lox) strain will be valuable for conditional Cpr gene deletion and subsequent determination of the impact of CPR loss on the metabolism of endogenous and xenobiotic compounds, as well as on postnatal development and other biological functions.

Alleles↗

Development and use of a rat albumin cDNA clone to evaluate the effect of chronic ethanol administration on hepatic protein synthesis.

A rat albumin cDNA probe (pBR alb 149) was developed in order to investigate the molecular mechanisms responsible for changes in hepatic protein synthesis after chronic administration of ethanol to rats. Rats fed a diet for up to 1 year in which 36% of calories were from ethanol, developed fatty livers but not cirrhosis. Cell-free protein synthesis with liver membrane-bound polysomes of ethanol-fed rats was increased as compared to control membrane-bound polysomes, whereas protein synthesis with free polysomes was unchanged. Total RNA extracted from liver membrane-bound polysomes and translated in a rabbit reticulocyte mRNA-dependent system showed a marked increase in albumin synthesis in the ethanol-fed group. Analysis of RNA molecules separated according to molecular weight by gel electrophoresis and hybridized with recombinant-cloned albumin cDNA demonstrated an increase in full-sized albumin mRNA species in ethanol-fed animals. Therefore, chronic ethanol administration appears to increase albumin synthesis by increasing the steady-state level of biologically active albumin mRNA in liver membrane-bound polysomes. Despite development of fatty liver, the protein synthesis machinery functions normally.

Albumins↗

Cell lines with heterogeneous phenotypes result from a single isolation of albumin-sv40 T-antigen transgenic rat hepatocytes.

Several immortalized cell lines were established from the livers of two transgenic rats expressing the simian virus protein large T antigen under the control of the albumin promoter. Hepatocytes from transgenic rats were isolated by a two-step perfusion procedure from a normal-appearing liver and from a liver that contained a single neoplasm. Cells were also isolated from the dissociated liver neoplasm. After 5 weeks in culture, cell colonies were isolated and subcloned as individual cell lines. Electron microscopy revealed that all cell lines had a high nuclear-to-cytoplasmic ratio compared with normal hepatocytes. The cytoplasm contained numerous organelles, including smooth and rough endoplasmic reticulum, Golgi, mitochondria, peroxisomes, and annulate lamellae. However, the lines exhibited a variety of different cell morphologies. All cell lines, including those derived from neoplastic cells, exhibited a similar doubling time of 26 hours and the ability to grow in soft agar. Northern blot analysis revealed that the cell lines differentially expressed hepatocyte markers. Large T antigen was expressed in the cultured cell lines at much higher levels than was observed in transgenic hepatocytes in vivo. This suggests that the viral protein is required to maintain cell viability in culture. In addition, the tumor suppressor proteins, p53 and Rbp105, were detected in all cultured cell lines. In contrast, these same proteins were not detected by Western blot in transgenic hepatocytes in vivo. All cell lines expressed the oncogene c-myc, yet growth factor-dependent and independent growth were observed. The data presented show for the first time the establishment and characterization of a number of cell lines derived from hepatocytes isolated from an alb-SV40 large T-antigen transgenic rat. These cell lines exhibited varied morphological and biochemical hepatocellular characteristics in vitro, suggesting that the expression of the transgene in hepatocytes leads to considerable phenotypic diversity analogous to that seen in hepatocellular carcinomas induced by chemical carcinogenesis.

Animals↗

Cellular and humoral anti-tumor immune responsiveness in chickens bearing tumors induced by avian sarcoma virus.

A systematic comparison was undertaken of the respective abilities of normal chicken embryo fibroblast (CEF) cells, Rous sarcoma virus (RSV)- transformed CEF cells, avian Rous sarcoma (RS) tumor cells and murine RS cells to serve as targets and antigen donors in various assays for the detection of cellular and humoral anti-tumor immunity in chickens bearing tumors induced by Rous sarcoma virus. As measured by a cytotoxicity procedure, avian and murine RS cells were more susceptible to the killing effects of sensitized lymphocytes than were transformed CEF which in turn were more reactive than normal CEF. In contrast, sera from tumor-bearing animals were able to stain by indirect immunofluorescence only the avian RS and transformed CEF cell types. Extracts of both transformed CEF cells and avian RS cells but not normal CEF were equally effective as inhibitors of migration of peritoneal exudate cells derived from tumor-bearing animals. Transformed CEF were found to produce far higher quantities of transforming virus progeny than avian RS cells, although the latter were apparently albe to synthesize defective viral particles. These data indicate the significant variations which may occur under experimental conditions, depending on the types of assays and target cells employed.

Animals↗

Specific uptake of alpha-fetoprotein by malignant human lymphoid cells.

We have studied the ability of human B- and T-lymphoblastoid cell lines, as well as of peripheral lymphoid cells from leukemia patients, to take up alpha-fetoprotein (AFP) and other serum proteins. Two technical approaches have been employed, both using fluorescent protein derivatives (FITC-proteins): microscopic examination of labelled cells using epifluorescent illumination and quantitation of endocytosed proteins by fluorescence-activated cell sorting (FACS). Compared to human resting T-lymphocytes, all T- and B-cell lines tested exhibited positive staining for fluoresceinated AFP and transferrin (Tf) and a significant increase, up to 100-fold, of the number of AFP and Tf molecules endocytosed per cell. Labelling was prevented or strongly diminished by a 100-fold excess of unlabelled protein. Preliminary results with peripheral lymphoid cells harvested from leukemia patients showed the presence of AFP- and Tf-positive cells in the blood of all patients examined. Intensity of labelling was related to the type of leukemia cells and/or the degree of cell maturation. Most cell lines exhibited positive staining for alpha 2-macroglobulin (alpha 2M) and also, to a lesser extent, for serum Vitamin D3 binding protein (DBP). In contrast, no labelling was observed with FITC-serum albumin (FITC-Alb) or FITC-ovalbumin (FITC-OVA). Comparative uptake of several FITC-proteins by a single cell population revealed significant quantitative and qualitative differences.

Blood Proteins↗

Surface field of forces and protein adsorption behavior of poly(hydroxyethylmethacrylate) films deposited from plasma.

Polymeric films were deposited from hydroxyethylmethacrylate (HEMA) plasma on non-woven poly(butyleneterephtalate) (PBT) filter materials. To test the effect of deposition conditions on surface properties, film were deposited using a constant monomer flow rate and a discharge power ranging from 40-100 W. Surface composition and surface energetics were evaluated by Electron Spectroscopy for Chemical Analysis (ESCA) and contact angle measurement, respectively. Albumin (Alb) and fibrinogen (Fg) adsorption from single protein solutions to the plasma-coated filters was measured. Results illustrate the marked effects of the deposition condition on the surface composition, the surface field of forces, and the protein adsorption behavior. The latter is modeled by the application of the Good-van Oss-Chaudhury theory of Lewis acid-base contribution to interfacial energetics. Materials endowed with widely different properties are obtained from the same monomer and different deposition conditions, a result that must be taken into account both in the production step, to assure constant quality, and in the development of specifically tailored materials.

Adsorption↗