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PubMed1982

Development of a two-step enrichment procedure for recovery of Yersinia enterocolitica from food.

Two new enrichment media were formulated for the recovery of Yersinia enterocolitica from foods: (i) yeast extract-rose bengal broth for preenrichment at 4 or 10 degrees C; and (ii) bile-oxalate-sorbose broth, a selective enrichment incubated at 22 degrees C. Comparison of these media in a two-step enrichment procedure against cold enrichment and modified Rappaport broth showed improved and more rapid recovery of human strains of Y. enterocolitica from inoculated foods. The use of bile-oxalate-sorbose broth as a selective enrichment also improved the performance of cold enrichment with phosphate-buffered saline. Determination of the best enrichment system for recovery of Y. enterocolitica from samples of retail pork and fresh pork tongues depended on whether the criterion was the number of positive samples, the variety of different serotypes recovered, or the ability to recover the important human serotype O:3. A single enrichment system with the widest selectivity would include preenrichment at 4 degrees C with either phosphate-buffered saline for 14 days or yeast extract-rose bengal broth for 9 days followed by selective enrichment with bile-oxalate-sorbose broth at 22 degrees C for 5 days.

Animals↗
PubMed1994

Incidence of apparent restricted diffusion in three different models of cerebral infarction.

High speed magnetic resonance imaging (MRI) and short diffusion times are used to investigate the appearance of restricted diffusion in three different models of cerebral infarction. The models are: the middle cerebral artery occlusion (MCAO) model in the rat, the carotid occlusion model in the gerbil, and the Rose Bengal microvascular occlusion model in the rat. All three were investigated for 16 b-values equally spaced between 10 and 1510 s/mm2 using two distinct experiments. In the ct (constant time) experiment, the diffusion time was held constant at 11.7 ms while the b-value was varied with the gradient strength. In the cg (constant gradient) experiment, the gradient strength was held constant and the b-value increased by varying the diffusion time from 4.4 to 11.7 ms. A monoexponential decay of the signal intensity with b-value in the ct experiment accompanied by nonmonoexponential (NME) decay in the cg experiment is indicative of restricted diffusion. As this phenomenon is detectable only at short diffusion times, it cannot be due to restriction by impermeable membranes, and we have thus termed this apparent restriction. For the MCAO model and the carotid occlusion model, apparent restriction was found both inside the infarct territory and in some regions outside it. No definite evidence for restriction was found for the Rose Bengal model, which was, however, only studied from 24 h post-insult.

Animals↗
PubMed2004

Anti-Ro/SSA and anti-La/SSB autoantibodies in the tear fluid of patients with Sjögren's syndrome.

PURPOSE: To investigate the presence of anti-Ro/SSA and anti-La/SSB antibodies in the tear fluid and serum of patients with Sjögren's syndrome and to evaluate the association of these autoantibodies with the severity of keratoconjunctivitis sicca. METHODS: Tear fluid and serum were obtained from 28 patients with Sjögren's syndrome and 17 age matched normal control subjects. Evaluation of tear fluid and sera anti-Ro/SSA and anti-La/SSB levels was done by using a quantitative enzyme linked immunosorbent assay kit designed for the quantitative measurement of IgG class autoantibodies directed against highly purified SSA and SSB antigens. Tear function and ocular surface were evaluated by Schirmer I test, tear break up time, and rose bengal staining. Dry eye symptom scores were recorded. RESULTS: Increased levels of anti-Ro/SSA and anti-La/SSB antibodies were detected in sera of 57.1% and 50% of SS patients, respectively. Six patients had increased levels of anti-Ro/SSA in the tear fluid, in one case anti-Ro/SSA being detected in tear fluid when it was negative in serum. Ten patients had positive anti-La/SSB titres in tear fluid and in four of these patients, anti-La/SSB titres were not elevated in serum. A positive correlation was observed between serum and tear fluid titres of anti-Ro/SSA (r = 0.43, p = 0.02), but not of anti-La/SSB. Serum anti-Ro/SSA and anti-La/SSB concentrations correlated positively with dry eye symptom scores (r = 0.42, p = 0.02 and r = 0.48, p = 0.01, respectively) and negatively correlated with Schirmer I test scores (r = -0.39, p = 0.04 and r = -0.40, p = 0.03, respectively). Significant correlations were found between tear anti-La/SSB concentrations and dry eye symptom score (r = 0.56, p = 0.02) and also rose bengal staining scores of the ocular surface (r = 0.44, p = 0.02). CONCLUSION: This study shows that autoantibodies against Ro/SSA and La/SSB antigens are present in the tear fluid of some patients with SS and their presence in serum or tear fluid is associated with the severity of keratoconjunctivitis sicca. Additional measurement of tear fluid levels of anti-Ro/SSA and anti-La/SSB may serve as a valuable diagnostic indicator of SS.

Adult↗
PubMed1993

Radiation hepatology of the rat: parenchymal and nonparenchymal cell injury.

Cellular changes that occur within the liver before and during liver fibrosis have been studied. At various times (0-90 days) after gamma irradiation (0-25 Gy) of the whole liver, hepatocytes and liver nonparenchymal cells were isolated by enzymatic dissociation of the liver and differential centrifugation. Differential cell counts were done to quantify the yield of hepatocytes and nonparenchymal cells. Hepatocyte function in vitro was assayed by the uptake of rose bengal and compared with the clearance of the dye in vivo. Intracellular alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were used to estimate the effect of radiation on the phenotypic expression of hepatocytes. Gene expression of transforming growth factor beta 1 (TGF-beta 1), a cytokine which has been found to play a role in hepatic fibrosis, was measured by 32P-cDNA:mRNA hybridization of poly(A+) mRNA isolated from purified populations of hepatocytes and nonparenchymal cells. Morphologically, hemorrhages were evident in perfused livers 28 days after 25 Gy whole-liver irradiation. This injury was preceded by a significant increase in liver nonparenchymal cells at 2 weeks, the earliest time after irradiation at which measurements were made. Significant increases in liver nonparenchymal cells occurred at doses greater than 10 Gy and persisted throughout the follow-up period after irradiation. The lowest dose studied (5 Gy) inhibited both normal growth of the liver and the associated increase in the number of hepatocytes. Doses greater than 10 Gy resulted in a dose-dependent decline in liver mass and hepatocytes. Viable hepatocytes isolated from irradiated dysfunctional livers, with respect to clearance of rose bengal, were functionally normal in vitro. A dose-dependent decline in the ALT and AST contents of liver cell suspensions and homogenates was observed but the relative decline in ALT was greater, resulting in lower ALT/AST ratios. This decline in the ALT/AST ratios occurred at doses (< 15 Gy) which caused no loss of hepatocytes, suggesting possible conversion of ALT-rich periportal hepatocytes to ALT-poor hepatocytes due to changes in the microenvironment and/or aging of the cell population. TGF-beta 1 mRNA was detected mainly in nonparenchymal cells, and radiation preferentially enhanced the TGF-beta 1 gene expression in these cells. These data suggest that radiation-induced alterations in liver nonparenchymal cell populations may be responsible for microvascular fibrosis, which results in a cascade of pathological events which lead to hepatocyte loss.

Alanine Transaminase↗
PubMed1999

Pb2+ promotes lipid oxidation and alterations in membrane physical properties.

Experimental evidence suggests that cellular damage mediated by oxidants could be involved in the pathology associated with lead (Pb) toxicity. We investigated the effect of Pb2+ on lipid oxidation in liposomes using different initiators. In the presence of Fe2+, Pb2+ (12.5-200 microM) stimulated lipid oxidation in phosphatidylcholine:phosphatidylserine-containing liposomes, measured as 2-thiobarbituric acid-reactive substances (TBARS) and conjugated dienes. This stimulatory effect depended on the presence of membrane negative charges and on bilayer integrity. Pb2+ did not stimulate TBARS formation in the presence of 25 mM 2,2'-azo-bis (2,4 dimethylvaleronitrile (AMVN) and 2,2' azobis (2-amidinopropane) (AAPH). Pb2+ significantly stimulated TBARS production and NADH oxidation in the presence of photoactivated rose Bengal. The use of specific inhibitors indicated that several reactive oxygen species were involved in the pro-oxidant action of Pb2+. Pb2+ (12.5-200 microM) caused membrane lateral phase separation and this effect was positively correlated with its capacity to stimulate Fe2+ and rose Bengal-initiated TBARS production. Pb2+ could bind to the membrane and act to stimulate lipid oxidation by causing changes in membrane physical properties. Through this mechanism Pb2+ would favor the propagation of lipid oxidation. By causing lateral phase separation and/or by increasing lipid oxidation rates, Pb2+ could be cytotoxic by altering membrane-related processes.

Amidines↗
PubMed1995

Melatonin protects neurons from singlet oxygen-induced apoptosis.

Singlet oxygen (O2[1 delta g]) is a very reactive molecule that can be produced by living cells and may contribute to cytotoxicity. The pineal hormone melatonin has been reported to possess potent antioxidant activity, and to be capable of scavenging O2(1 delta g). We investigated whether melatonin might reduce the neurotoxic action of O2(1 delta g). The cytotoxic effect of singlet oxygen was studied in primary cultures of cerebellar granule neurons pretreated with a photosensitive dye, rose bengal, and exposed to light--a procedure that generates O2(1 delta g). We found that this procedure triggers neuronal death, which is preceded by mitochondrial impairment (assayed by the rate of the reduction of MTT, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide, into formazan), and by DNA fragmentation--a marker of apoptosis. DNA fragmentation was determined in situ by terminal deoxynucleotidyl transferase assay; cell death was assayed with 0.4% trypan blue solution--viable cells with an intact membrane are not permeable to trypan blue; dead cells are, and thus, they are stained blue. Neuroprotection was obtained with the pineal hormone melatonin. In a cell-free system, melatonin also protected the enzyme creatine kinase (EC 2.7.3.2) from the rose bengal-induced injury. The results suggest that melatonin might counteract the cytotoxic action of singlet oxygen. Further studies are needed to clarify the exact role singlet oxygen and melatonin might play in neurodegenerative diseases.

Animals↗
PubMed1970

The interaction of triethyltin with a component of guinea-pig liver supernatant. Evidence for histidine in the binding sites.

A protein fraction was isolated from guinea-pig liver that binds triethyltin with an affinity of approx. 2x10(6)m(-1) at pH8.0. It was shown that the protein responsible for binding 70% of the triethyltin found in guinea-pig liver after injection of radioactively labelled triethyltin is at most a few per cent of the total liver protein. Evidence is presented from the kinetics of loss of binding and loss of certain amino acids on photo-oxidation with either Methylene Blue or Rose Bengal that each binding site consists of two histidine residues.

Animals↗
PubMed1975

Phenobarbital effects in cholestatic liver diseases.

Fifteen patients with cholestatic disorders were treated for 1 to 5 months with phenobarbital. Primary biliary cirrhosis was diagnosed in seven, sclerosing cholangitis in two, intrahepatic biliary hypoplasia in three, and cholestatic hepatitis in three. Except for the patients with cholestatic hepatitis, in whom marked cholestasis was virtually the only abnormality in liver biopsy specimens, serum bilirubin and bile acid concentrations were diminished during therapy, the hepatic clearance of sulfobromophthalein and 131-I-rose bengal was variably enhanced, and there was relief from pruritus. Serum cholesterol concentrations and other measures of hepatic function were not significantly changed during therapy except for serum alkaline phosphatase activity, which rose in twelve patients. Parallel changes occurred in 5'-nucleotidase, suggesting a hepatic origin for the alkaline phosphatase activity. These studies indicate that phenobarbital therapy is associated with improvement in organic anion clearance in some patients with cholestatic disorders and may be beneficial to such patients.

Adult↗
PubMed1981

A flow-cell method for measuring hepatic excretion of pigments continuously in rats.

The electronic absorption of effluent bile from restrained rats with indwelling bile cannulas can be measured directly and continuously in the wavelength region 430-800 nm using commercially available micro flow-cells. The method is useful for studying and characterizing the hepatic excretion of endogenous and exogenous pigments. Biliary excretion curves are obtained directly without requiring separate analyses of individual bile samples. To demonstrate its application and scope, the technique has been used to follow the biliary excretion of sulfobromophthalein, rose bengal, biliverdin, and bilirubin.

Animals↗
PubMed1980

Intrahepatic gallbladder.

Intrahepatic gallbladder must be distinguished from other lesions which also produce an area of decreased tracer concentration on liver scan. Identification is imperative because it determines therapy. A scan performed with rose bengal labeled with iodine 131 was used in this case.

Gallbladder↗
PubMed1987

The effect of a surfactant in media for the enumeration, growth and identification of airborne fungi.

Air sampling on to culture media for viable fungi is complicated by the presence of fast-growing species. A non-ionic surfactant (Triton N-101) incorporated into the medium effectively reduced the spread of such species and thus facilitated the enumeration and identification of fungal colonies collected by an Andersen sampler. The concentration of surfactant from 250 to 8000 ppm was not critical. Sixteen species of common moulds inoculated on surfactant media showed restricted radial growth but never total inhibition. Surfactant did not prevent sporulation and did not greatly affect morphological characteristics. Rose bengal media performed poorly in comparative tests. These observations indicate the practical value of this surfactant in aeromycological studies.

Air Microbiology↗
PubMed1978

Photooxidation of soybean apoleghemoglobin with protoporphyrin IX, heme and dye.

Soybean apoleghemoglobin a was irradiated with visible light in the presence of different sensitizers to probe the heme environment of the protein. With protoporphyrin IX as sensitizer, specific photooxidation of histidine-92 and histidine-61 occurred. Irradiation of oxyleghemoglobin and cyanleghemoglobin resulted in photooxidation of histidine-92, while addition of methylene blue caused both histidine-92 and histidine-61 to be oxidized. Apoleghemoglobin, irradiated in the presence of rose bengal or methylene blue, lost tryptophan-128 in addition to the two histidines.

Apoproteins↗
PubMed1975

A mechanism for prozone formation in the complement fixation test for bovine brucellosis.

Complement fixation (CF) by bovine IgG1 or IgM antibodies to Brucella abortus was inhibited by specific non-complement-fixing antibodies of the IgG2 subclass. This inhibition may account for the appearance of prozones in CF titrations of some antiserums, and for the occurrence of serums which are positive to the Rose Bengal test, but negative to the CF test.

Agglutination Tests↗
PubMed1991

Beneficial effect of sodium sucrose-sulfate on the ocular surface of patients with severe KCS in primary Sjögren's syndrome.

Sucralfate (aluminium sucrose-sulfate), a well known gastric mucosal protectant, has been tested topically on 22 patients (20 females and 2 males) suffering from primary Sjögren's syndrome. Median treatment period was 6 months (range 1-19 months). Statistically significant improvement in the ocular surface condition was found judged from the reduction in Rose-Bengal score (P less than or equal to 0.00005). The beneficial effect appeared within the first 1-4 months of treatment. No adverse side effects were encountered.

Administration, Topical↗
PubMed1998

Micropachometric quantification of tear assessment tests.

After reviewing the results of available methods for defining the dry eye with an exact assessment of severity, we found that micropachometry was the most sensitive. The following is a ranking of the other tests according to usefulness: Tear resistance, Schirmer, and BUT. Rose bengal stained eyes were usually decompensated; the meniscus test's primary use was as an "all or nothing" indicator, but recording the widths of extant menisci were unproductive.

Coloring Agents↗
PubMedDate not supplied

Eye parameters for the diagnosis of xerophthalmos.

The anatomic aspects of the lacrimal apparatus are reviewed in relation to tear production, discussing the concept of xerophthalmos, its etiology and tests for its diagnosis. The authors use two diagnostic tests which quantify the serous secretion (Schirmer's or Jones's tests) and demonstrate epithelium damage (fluorescein or rose Bengal dye test). In cases of presumed xerophthalmos, other tests such as determination of lactoferrin, muramidase, IgA, TISC/BUT, etc. may also be performed to confirm the diagnosis.

Endothelium, Corneal↗
PubMed1977

Structural rearrangements due to ligand binding and haem replacement in myoglobin and leghaemoglobins.

Structural rearrangements in sperm whale myoglobin and leghaemoglobins caused by changes in the spin or oxidation state of the iron as a consequence of ligand binding have been measured by difference spectroscopy in the ultraviolet. When compared with the high-spin acetate complex, ligands which cause a transition to the low-spin state also cause large perturbations of tyrosine(s) remote from the haem pocket in myoglobin but only minor perturbations of tryptophan (s) in leghaem calobin. This may indicate a weaker coupling between events at the haem site and conformational changes in the protein in leghaemoglobins. The absorption spectra of various haem-liganded forms of the two proteins as well as the binding of the dye rose Bengal to the two apoproteins are consistent with weaker interactions between the haem ano apoprotein and a more solvent-exposed haem pocket in leghaemoglobin compared with myoglobin.

Animals↗