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Y Ni

Publications and source records attributed to Y Ni.

At least 91 records · Page 5Linked to original sources

Studies on unusual cytoplasmic structures which contain rabies virus envelope proteins.

We investigated unusual structures produced in BHK-21 cells infected with rabies virus (HEP-Flury strain). Sellers' staining of the cells revealed, in addition to Negri body-like structures (inclusion bodies), production of a fuchsin-stained cytoplasmic structure (FCPS) which encircled the nucleus. The frequency of the FCPS-forming cells increased as replication progressed. The FCPS was different from the inclusion body because the former contained the viral glycoprotein (G) and matrix protein (M2) antigens, while the latter contained nucleocapsid antigens. In the early phase of infection, we observed accumulation of viral envelope antigens in a cytoplasmic structure that was considered to be expanded rough endoplasmic reticulum (rER) because of its concomitant increase in BiP content. Time-course studies suggested that the envelope antigen-containing structure, which was not stained with basic fuchsin, translocated to the perinuclear region to form the FCPS. FCPS formation was dependent on incubation temperature and was decreased at 30 degrees C, while the development of virus-induced cytopathic effect (CPE) was delayed. When the incubation temperature was shifted up to 37 degrees C, FCPS formation was induced again and progression of CPE was accelerated in approximate proportion to the increasing number of FCPS-positive cells. From these studies, we conclude that viral G proteins gradually accumulate in the rER with M2 protein and the expanded rER converts eventually into the FCPS, which may be closely related to accelerated host cell death.

Animals↗

Recent developments in pharmacokinetic modeling of perfluorocarbon emulsions.

Perfluorocarbon (PFC) emulsions are potential oxygen carriers. This study is to investigate pharmacokinetic compartment models and physiologically based models, which correlate the distribution of PFC emulsion in the blood, reticuloendothelial system (RES) tissues and non-RES tissues with the excretion data. The models are evaluated by nonlinear regression analysis (using PCNONLIN software) with expiration data from animal following an i.v. injection of a concentrated perflubron emulsion. One model with four compartments (representing PFC emulsion in blood, RES tissues, non-RES tissues, and PFC solubilized in blood) meets the mathematical and physical criteria. The physiological modeling provides insight of physiological mechanisms. The relationship between the compartment model parameters (rate constants) and physiological parameters (tissue volumes, flow rates, etc.) is presented. An advantage of physiological model is that prediction may be made in interspecies scaling. The above two kinds of modeling are useful in many applications, e.g. to describe and predict the time course of PFC disposition throughout the body.

Animals↗

Effect of manganese dipyridoxal diphosphate on liver magnetic resonance imaging and serum bilirubin in rats with removable biliary obstruction.

RATIONALE AND OBJECTIVES: It is known that manganese dipyridoxal diphosphate (Mn-DPDP) causes persisting liver enhancement in cholestatic rats, that free Mn++ plus bilirubin induces intrahepatic cholestasis, and that free Mn++ is released in vivo after Mn-DPDP injection. Hence, there is a concern about potential secondary intrahepatic cholestasis in patients who have biliary obstruction. In this study, we further investigated this issue. METHODS: Removable total biliary obstruction (RTBO) was induced in 12 rats. Six of them (group A) received Mn-DPDP (25 mumol/kg). The others (group B) served as control animals. The data from serial magnetic resonance imaging and serum bilirubin tests were compared. RESULTS: Without Mn-DPDP, a minimal increase of the liver intensity was observed in both groups because of cholestasis. In group A, the intensity of the liver was strongly enhanced with Mn-DPDP but normalized within 48 hr after removal of the obstruction. In both groups, total bilirubin levels increased up to 131.67 mumol/l 2 days after RTBO but rapidly decreased within 4 hr and almost normalized within 24 hr after removal of the obstruction, suggesting a lack of Mn-DPDP influence on the bilirubin level. CONCLUSION: We found that Mn-DPDP did not cause secondary intrahepatic cholestasis. Retained Mn++ is likely eliminated after restoration of bile flow. These results indicate that Mn-DPDP can be used in patients who have obstructive jaundice as long as it is followed by successful bile drainage.

Animals↗

Localization of metalloporphyrin-induced "specific" enhancement in experimental liver tumors: comparison of magnetic resonance imaging, microangiographic, and histologic findings.

RATIONALE AND OBJECTIVES: We investigated the tumor specificity of gadolinium mesoporphyrin (Gd-MP) and manganese tetraphenylporphyrin (Mn-TPP) as magnetic resonance (MR) imaging contrast agents. METHODS: Fifteen rats with multiple hepatocellular carcinomas and eight rats with implanted Novikoff hepatomas were given intravenous injections of either Gd-MP or Mn-TPP at 0.05 mmol/kg, which was compared with nonspecific gadopentetate dimeglumine (0.3 mmol/kg). T1-weighted spin-echo images were obtained before and up to 48 hr after injection and compared with corresponding microangiograms and histologic specimens. The relative enhancement of organs and tumors was plotted as a function of time. RESULTS: Initially, both metalloporphyrins behaved as nonspecific agents, similar to gadopentetate dimeglumine, and enhanced the tumor by perfusion and diffusion. However, metalloporphyrins, but not gadopentetate dimeglumine, caused a delayed (> or = 3 hr) enhancement in some compartments of certain lesions. The MR imaging-microangiography-histology matching technique revealed that those compartments were actually nonviable components, including necrosis (n = 10), thrombosis (n = 7), and cystic secretion (n = 3), but not viable tumor tissue. CONCLUSION: Metalloporphyrins did not prove to be tumor specific. However, the observed affinity for nonviable tissue has elicited other potential applications for these agents.

Angiography↗

Mapping and characterization of a sequential epitope on the rabies virus glycoprotein which is recognized by a neutralizing monoclonal antibody, RG719.

We have established a murine hybridoma cell line RG719 which produces a rabies virus-neutralizing IgM-type monoclonal antibody (referred to as MAb RG719). Immunoblot analysis indicated that the antibody recognized a sequential epitope of G protein. Among four rabies virus strains tested, the antigenicity to MAb RG719 was absent from the Nishigahara strain, while the other three strains (HEP, ERA and CVS) reacted to the MAb. Studies with deletion mutants of the G protein indicated that the epitope was located in a middle region of the primary structure of G protein, ranging from position 242 to 300. By comparing the estimated amino acid sequence of the four strains, we found in this region two amino acids (at positions 263 and 291) which are common to three of those strains but are not shared by the Nishigahara strain. The site-directed point mutagenesis revealed that replacement of phenylalanine-263 by leucine destroyed the epitope of the HEP G protein, while the epitope was generated on the Nishigahara G protein whose leucine-263 was replaced by phenylalanine. These observations suggest that phenylalanine-263 is essential for constructing the epitope for MAb RG719. The synthetic 20-mer peptide produced by mimicking the amino acid sequence (ranging from amino acid positions 249 to 268) of the presumed epitope region was shown to bind specifically to MAb RG719 and also to raise the virus-neutralizing antibodies in rabbits. Vaccination with the HEP vaccine produced in Japan induced in humans and rabbits production of significant amounts of the antibodies which reacted with the 20-mer peptide.

Amino Acid Sequence↗

Potential role of bile duct collaterals in the recovery of the biliary obstruction: experimental study in rats using microcholangiography, histology, serology and magnetic resonance imaging.

Obstructive cholestasis induced in animals at the level of the lobar and common bile ducts is known to be reversible with time. This study was conducted not only to test the hypothesis that formation of bile duct collaterals is responsible for the recovery of biliary obstruction but also to assess the potential of hepatobiliary agent-enhanced magnetic resonance imaging for visualizing cholestasis. A total of 52 rats were divided into three groups with selective biliary obstruction, total biliary obstruction and sham surgery. We studied the evolution of cholestasis by correlating microcholangiographic, histological findings with the results of liver tests and hepatobiliary agent-enhanced magnetic resonance imaging. Lobar cholestasis undetected by liver tests but seen on magnetic resonance imaging as a difference between ligated and unligated lobes, occurred in 15 out of 20 rats subjected to selective biliary obstruction within 48 hr after ligation, and recovered later on as a result of the development of bile duct collaterals. Five rats failed to show local cholestasis as a result of the existence of interlobar accessory bile channels. All 18 total biliary obstruction-treated rats were cholestatic soon after ligation, as confirmed by high serum bilirubin and alkaline phosphatase levels and as documented by poor liver enhancement on magnetic resonance imaging. Cholestasis recovered within 4 wk with normalization of liver enhancement on magnetic resonance imaging as a result of the formation of bile duct collaterals (as demonstrated by microcholangiographic and histological study). Bile duct collateral formation is responsible for the recovery from obstructive cholestasis in rats. A similar mechanism might be present in conditions of bile duct obstruction without cholestasis. Hepatobiliary agent-enhanced magnetic resonance imaging is more sensitive than blood tests in detecting local cholestasis and can be used to monitor noninvasively the evolution of biliary obstruction.

Alanine Transaminase↗

Subgenomic S1 segments are packaged by avian reovirus defective interfering particles having an S1 segment deletion.

Two subgenomic segments derived from the S1 genome segment (SGS1-1 and SGS1-2) were identified in avian reovirus defective interfering (DI) particle preparations having an S1 segment deletion. Both SGS1-1 and SGS1-2 were composed of double-stranded RNA (dsRNA) with an estimated size of 400 and 380 bp, respectively. Their segment of origin was identified as the S1 by hybridization analysis. The subgenomic segments were associated with the virus fraction following CsCl density gradient centrifugation, indicating that they are packaged. The subgenomic segments were also shown to be replicated. Therefore, sequence(s) required for replication and packaging are retained. The relative amounts of subgenomic segments were shown to be inversely proportional to that of the S1 segment. The presence of subgenomic segments and concurrent reduction in the relative amount of the S1 segment were found to be directly associated with the decrease in infectious titers. These results suggest that subgenomic segments are responsible for induction of interference by specifically competing with the S1 segment during replication and/or packaging. The competitive relationship between the subgenomic segments and the S1 segment implies that segment-specific sequence(s) or factor(s) are involved in the replication and/or packaging of each individual genome segment.

Animals↗

Clinical observation of treatment of hypertension with calcium.

The effect of oral calcium supplementation (1000 mg/day) on hypertension was studied in 57 borderline and mild-to-moderate hypertensive patients in a randomized, double-blind, placebo-controlled study for 14 weeks. Twenty-five patients from the above groups (11 from the calcium-treated group and 14 from the placebo group) were studied in a crossover fashion for 14 more weeks. The high calcium intake lowered systolic blood pressure by 17 mm Hg (P < .01), and diastolic blood pressure by 11 mm Hg (P < .01). Fifty percent of the calcium-treated patients showed a significant antihypertensive effect and were termed calcium responders. In the crossover study, serum sodium was lower after taking calcium than after placebo intake (P < .05). Pretreatment plasma free calcium content of the calcium-responsive patients was significantly lower (P < .05) than in the calcium nonresponsive patients, and was highly significantly increased (P < .01) after administering calcium. The result showed that oral calcium supplementation can lower blood pressure in a significant fraction of essential hypertensive subjects, and that the free calcium level in plasma may help identify calcium-responsive individuals. While the mechanism by which increased calcium intake lowers blood pressure in hypertension is still undetermined, these data support an underlying relationship between hypertension and calcium and possibly sodium metabolism.

Administration, Oral↗

MR imaging evaluation of liver enhancement by Gd-EOB-DTPA in selective and total bile duct obstruction in rats: correlation with serologic, microcholangiographic, and histologic findings.

PURPOSE: To investigate the behavior and potential application of the hepatobiliary magnetic resonance (MR) imaging contrast agent gadolinium-ethoxybenzyl (EOB)-diethylene-triamine-pentaacetic acid (DTPA) in cholestasis. MATERIALS AND METHODS: Rat models of selective biliary obstruction (SBO) (n = 20) and total biliary obstruction (TBO) (n = 14) were used. Serial Gd-EOB-DTPA-enhanced MR images were obtained, and the findings were correlated with serologic, microcholangiographic, and histologic findings. RESULTS: In SBO rats, statistically significantly prolonged enhancement of ligated lobes occurred during the acute phase (P < .01) but was absent during the chronic phase owing to bile collateral vessels. In TBO rats, statistically significantly lower liver enhancement (P < .01) was observed as long as the obstruction persisted, until collateral bile ducts developed. CONCLUSION: The distinction between obstructed and unobstructed liver on Gd-EOB-DTPA-enhanced MR images suggests a novel application for visualizing cholestasis.

Animals↗

Effects of buffer pH and phosphate concentration on the droplet size and EYP hydrolysis of perflubron/EYP emulsions.

Oil-in-water emulsions containing perflubron (perfluorooctyl bromide; PFOB) and stabilized with egg yolk phospholipid (EYP) have potential applications as contrast agents and oxygen carriers. In this study, the effects of buffer pH and total phosphate concentration on the emulsion droplet size and EYP hydrolysis were evaluated. 90% w/v perflubron emulsions with NaH2PO4-Na2HPO4 buffers of different pH (4.7-8.7) and phosphate concentrations (30 and 60 mM) were prepared with a high-pressure homogenizer. Emulsions were stored at 40 degrees C and tested at 0, 1, 2 and 3 months. The pH dropped quickly in emulsions with pH 8.7 buffer whereas acidic and neutral buffered emulsions exhibited minor pH drops. The concentration of free fatty acids (FFA) vs emulsion pH can be fitted to a parabolic curve with a minimum at about pH 6.0. The droplet growth rates in emulsions with the pH 4.7 buffer were about 2.5 times of those in emulsions with the pH 8.7 buffer. Total phosphate concentration had only a minor effect. This study emphasizes the importance of the careful selection of buffer pH and capacity to control EYP hydrolysis and possibly emulsion droplet size.

Buffers↗

Effects of formulation, processing and storage parameters on the characteristics and stability of perflubron emulsion.

In this study, the effects of formulation, processing and storage parameters on perflubron (perfluorooctyl bromide; PFOB) emulsions were investigated. Emulsions with varying concentrations of perflubron and egg yolk phospholipid (EYP) were prepared with different processing parameters and placed at different storage temperatures. Their characteristics and stability were compared. The emulsion droplet growth rate was nearly proportional to the perflubron percentage in the range of 15-110% w/v. The initial droplet size of perflubron emulsions was inversely proportional to the concentration of EYP until a certain lower limit of droplet size was reached. The initial droplet size and droplet growth rate of perflubron emulsion were strongly dependent upon the processing parameters. The logarithmic value of the droplet growth rate decreased linearly with l/T in the range of 5-40 degrees C. The formulation and processing parameters are the key variables to be optimized to achieve better emulsion characteristics and stability.

Drug Stability↗

Inhibitors of sterol synthesis. Tritium-labeled 26-hydroxycholesterol of high specific activity from a byproduct of the Clemmensen reduction of diosgenin.

(25R)-26-Hydroxycholesterol (I) was synthesized in six steps from (22Z,25R)-cholesta-5,22-diene-3 beta,26-diol (II) in 31% overall yield. The 26-tert-butyldiphenylsilyl ether of II was converted via its 3 beta-tosylate to (22Z,25R)-6 beta-methoxy-26-(tert- butyldiphenylsilyloxy)-3 alpha,5-cyclo-5 alpha-cholest-22-ene (V). Removal of the 26-silyl group of V gave (22Z,25R)-6 beta-methoxy-3 alpha,5-cyclo-5 alpha-cholest-22-en-26-ol, which was hydrogenated over platinum oxide and then hydrolyzed to I. Catalytic reduction in the presence of deuterium or tritium gas gave [2H]-I or [3H]-I, respectively. Analysis of the [2H]-I by mass spectrometry showed that all the deuterium was located in the sterol side chain, mainly as d2, d3, and d4 species. The 2H and 13C nuclear magnetic resonance (NMR) spectra of [2H]-I indicated that most of the deuterium was located at C-22 and C-23, with lesser amounts at C-24 and minor amounts at C-20, C-21, C-25, and C-27. NMR spectra of [2H]-I and its alpha-methoxy-alpha-(trifluoromethyl)phenylacetate diester showed no detectable 20S epimer and approximately 2% of the 25S epimer. The [3H]-I was prepared analogously to [2H]-I using carrier-free tritium and showed a specific activity of 16.9 Ci/mmol. All synthetic intermediates were characterized fully by 1H and 13C NMR, and representative 1H-1H coupling constants are given for the ring A protons of i-steroids.

Deuterium↗

Identification of proteins encoded by avian reoviruses and evidence for post-translational modification.

Twelve avian reovirus strain 176 proteins, 10 structural (lambda 1, lambda 2, lambda 3, mu 1, mu 2/mu 2C, sigma 1, sigma 2, sigma 3, and sigma 4), and 2 nonstructural (mu NS and sigma NS) were identified and characterized by electrophoretic analysis of purified virions and infected cell lysates. Three of the identified proteins (mu 1, mu NS, and sigma 4) have not been previously described. In pulse-chase experiments, 10 of the proteins (lambda 1, lambda 2, lambda 3, mu 1, mu 2, mu NS, sigma 1, sigma 2, sigma NS, sigma 3) were shown to be primary translation products, whereas mu 2C was shown to be a post-translational cleavage product of mu 2.

Animals↗

Characterization of avian reovirus-induced cell fusion: the role of viral structural proteins.

Cell fusion induced by avian reovirus was analyzed using virus strain FC and Vero cells. One-step growth curves showed that cell fusion was directly associated with viral replication. Cell fusion occurred most efficiently at basic pH (8.0-8.5) and fusion from without could not be demonstrated. Actinomycin D, at low concentrations, increased cell fusion, and cycloheximide prevented cell fusion, indicating that viral protein(s) were responsible for the induction of cell fusion. Immunofluorescence tests indicated that viral proteins were present on the infected cell surface. Radioimmuno-precipitation identified structural proteins mu 2C and sigma 2 as predominant viral protein species present on the infected cell surface. Cell fusion was inhibited by virus-specific antisera, suggesting that mu 2C and/or sigma 2 present on the infected cell surface were involved in the induction of cell fusion. Trypsin and chymotrypsin treatment of purified viruses cleaved both mu 2C and sigma 2 proteins, but generated different cleavage products with each protein. The addition of trypsin to the culture media following infection increased cell fusion, whereas chymotrypsin treatment decreased cell fusion. The opposite effects of trypsin and chymotrypsin on the cell fusion, together with the different specificities of these two proteases in cleavage of mu 2C and sigma 2 proteins, further suggest that the cell surface-associated mu 2C and/or sigma 2 are involved in the syncytium formation.

Alkalies↗

Comparison between Gd-DTPA, Gd-EOB-DTPA, and Mn-DPDP in induced HCC in rats: a correlation study of MR imaging, microangiography, and histology.

The behaviour of two liver-specific contrast MR agents, Gd-EOB-DTPA and Mn-DPDP and one nonspecific contrast agent, Gd-DTPA, was compared in a rat model of chemically induced hepatocellular carcinoma (HCC). The study included contrast enhanced MR imaging and the corresponding microangiography and histology. Analysis of the MR images showed similar degrees of maximum relative liver enhancement: 47.5 +/- 8.2% for Gd-EOB-DTPA (0.03 mmol/kg) at 5 min postinjection and 52.5 +/- 14.4% for Mn-DPDP (0.025 mmol/kg) at 15 min; both exceeded the value obtained with Gd-DTPA (34.8 +/- 13.6%, at 5 min), even at 0.3 mmol/kg. Gd-EOB-DTPA caused a similar "negative" enhancement of all types of HCC, independent of their differentiation and vascularization, i.e., lesion-to-liver contrast-to-noise ratio (CNR) of differentiated and undifferentiated HCC increased negatively from, respectively, 1.9 +/- 1.1 and -5.1 +/- 3.1 before contrast to -5.2 +/- 2.4 and -11.8 +/- 4.8 at 5 min after contrast. On Mn-DPDP enhanced images, the undifferentiated HCCs showed up negatively (CNR -5.5 +/- 4.7 before contrast to -13.7 +/- 10 at 15 min after contrast), whereas the more differentiated tumors showed up positively (CNR from 2.3 +/- 2.0 before contrast to 12.5 +/- 3.5 at 24 hr postcontrast) due to active uptake and delayed elimination of Mn-DPDP.

Angiography↗

The uptake of manganese dipyridoxal-diphosphate by chemically induced hepatocellular carcinoma in rats. A correlation between contrast-media-enhanced magnetic resonance imaging, tumor differentiation, and vascularization.

OBJECTIVES AND RATIONALE: Negative enhancement of implanted liver tumors has been achieved in preclinical studies on manganese dipyridoxal-diphosphate (Mn-DPDP), a new hepatobiliary specific contrast agent for magnetic resonance imaging (MRI). The authors investigated the effects of Mn-DPDP on primary liver cancer and its possible mechanisms. METHODS: Magnetic resonance imaging was performed in 15 rats with chemically induced hepatocellular carcinoma (HCC) before and after Mn-DPDP injection. Both tumor-liver contrast-to-noise ratio and absolute tumor enhancement were evaluated and compared with the MRI results of a nonspecific contrast agent, gadolinium-DOTA, and correlated with corresponding microangiographic and histologic findings. RESULTS: Mn-DPDP injection led to a persistent positive enhancement in differentiated solid HCCs (22/23) with maximal conspicuity at 24 hours. Undifferentiated HCCs were all delineated by a prompt negative enhancement (20/20) with maximal conspicuity within 30 minutes.

Angiography↗

Experimental liver cancers: Mn-DPDP-enhanced rims in MR-microangiographic-histologic correlation study.

During both animal and clinical studies with manganese (II) N,N'-dipyridoxylethylenediamine-N,N'-diacetate 5,5'-bis(phosphate) (DPDP), enhanced rims around liver tumors on magnetic resonance (MR) images have been observed. To elucidate the origins of these rims and to assess their potential value in the differential diagnosis of liver masses, the authors studied 15 rats with induced hepatocellular carcinoma (HCC), 10 rats with implanted Novikoff hepatomas, and 11 rabbits with implanted VX2 carcinomas. A total of 69 primary and secondary liver cancers from these three animal models were studied. Mn-DPDP- and gadolinium tetraazacyclododecantetra acetic acid (DOTA)-enhanced MR images were compared. On the Mn-DPDP-enhanced images, 34 peritumoral rims of various patterns were displayed, all of which were exclusively related to the presence of highly malignant primary and secondary liver tumors. Peritumoral zones of malignant infiltration, surrounding parenchymal compression, and bile duct proliferation were seen to be the origin of these rims after comparison of MR images with the corresponding microangiograms and histologic specimens.

Angiography↗

Magnetic resonance imaging, microangiography, and histology in a rat model of primary liver cancer.

RATIONALE AND OBJECTIVES: N-nitrosodiethylamine is able to induce various benign and malignant liver lesions in rats with a high success rate and a low mortality rate. It provides a more appropriate model that better simulates the various lesions occurring in patients than the usual model of tumor implantations. METHODS: Hepatic carcinogenesis was induced in 58 Wistar rats using oral N-nitrosodiethylamine. The rats subsequently were studied by liver magnetic resonance imaging (MRI), postmortem microangiography, and histologic examination. RESULTS: Hepatic tumors developed in 57 rats. A wide variety of the tumors in the degree of vascularization, the type of vessels, the areas of intratumoral secretion and necrosis, and the tumor cell differentiation resulted from the tumor model. The authors were able to assess the contribution of the vascular, extravascular, and cellular components in the final pattern of contrast enhancement in MRI. CONCLUSIONS: The N-nitrosoethylamine model for hepatic tumor induction is simple, and provides a more representative range of tumors for experimental evaluation.

Animals↗