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Biomedical subjects

S Grossman

Publications and source records attributed to S Grossman.

At least 55 records · Page 3Linked to original sources
PubMed1995

Lipoxygenase activity in rat dermis and epidermis: partial purification and characterization.

Lipoxygenase (LOX) activity in epidermis and dermis was distributed among microsomal and cytosolic fractions. The main products of polyunsaturated fatty acid metabolism were 12-hydroperoxy-cis-5,8,14, trans-10-eicosatetraenoic acid (12-HPETE), 15-hydroperoxy-cis-5,8,11, trans-13-eicosatetraenoic acid (15-HPETE) and 13-hydroxy-cis-9, trans-11-octadecadienoic acid (13-HOD). Enzyme activities were isolated from rat dermis and epidermis by ammonium sulphate precipitation, hydrophobic chromatography and gel filtration. In the dermis, activity was found at a molecular mass of 68 kDa, a pI of 4.6 and a Km of 50 microM. This activity was inhibited by known LOX inhibitors. The main reaction products indicated that this was 15-LOX. In the epidermis, activity was found in a fraction with a molecular mass of 68 kDa, a pI of 4.6 and a Km of 80 microM. Activity was inhibited by known LOX inhibitors whereas the reaction products indicated that this was 12-LOX. LOX activity in rat skin may involve one enzyme with dual regional specificities or may comprise two different enzymes.

Animals↗
PubMed1995

Unilateral renal ischemia reperfusion in the rat: effect of blood volume trapped in the kidney, sucrose infusion, and antioxidant treatments.

This study was carried out in order to examine whether the severity of acute renal failure observed during the four hours following a 45 min period of unilateral occlusion of the renal pedicle could be reduced by various treatments. These include intrarenal flush with saline immediately before the occlusion, by sucrose infusion immediately before reperfusion, or by injection of NAO (natural antioxidant) and vitamin E before the occlusion. After renal pedicle occlusion, creatinine levels increased to 165% of their pre-ischemic values. Urine flow, GFR, renal cortex blood flow and NADH decreased by 99%, 99%, 50% and 36%, respectively. A decrease in the Na and K reabsorption (15% and 32%, respectively) was also observed. Partial protection of renal function against ischemic damage was observed when kidney tissue remained blood-free, by exposing it to saline throughout the period of ischemia. Significant protection was observed after treatment with sucrose, vitamin E and NAO. This study demonstrates that it is possible to attenuate the injury to the ischemic kidney by inducing ischemia in a bloodless kidney, by inducing diuresis in the first phase of reperfusion, or by antioxidant treatment, such as vitamin E or NAO.

Acute Kidney Injury↗
PubMed1995

Increased sensitivity to bilateral nephrectomy in rat caused by endotoxemia.

In this study the possibility that the kidneys play a role in the removal of endotoxin from the blood was examined. This was carried out by an IP injection of endotoxin (at a dosage of 1 mg/100 gr) to rats that had undergone bilateral nephrectomy, and examining the times of death following various treatments. Furthermore, the rats underwent treatment for endotoxin tolerance or received treatment with the antioxidants superoxide dismutase (SOD) and natural antioxidant (NAO) or were injected with an inhibitor of the enzyme NOS (NGmethyl l-arginine 1-NMA). The endotoxin concentration in the plasma was also measured. The results of this study indicate the rats were more sensitive to bilateral nephrectomy in the presence of endotoxin. This increased sensitivity in the binephrectomized rat decreases following treatment inducing tolerance to endotoxin, or following treatment with an antioxidant or with the inhibitor of NOS. A possible explanation for the increased sensitivity following endotoxin injection is that it apparently results from an increased concentration of endotoxin in the blood due to the impossibility of removing the endotoxin via the kidneys. Therefore, endotoxin tolerance or abolishing the potential injuries that may be caused by endotoxin negate the sensitivity in the binephrectomized rats injected with endotoxin.

Animals↗
PubMed1995

Epstein-Barr virus nuclear protein 3C modulates transcription through interaction with the sequence-specific DNA-binding protein J kappa.

The Epstein-Barr virus (EBV) nuclear protein 3C (EBNA 3C) is essential for EBV-mediated transformation of primary B lymphocytes, is turned on by EBNA 2, and regulates transcription of some of the viral and cellular genes which are regulated by EBNA 2. EBNA 2 is targeted to response elements by binding to the DNA sequence-specific, transcriptional repressor protein J kappa. We now show that EBNA 3C also binds to J kappa. EBNA 3C causes J kappa to not bind DNA or EBNA 2. J kappa DNA binding activity in EBV-transformed lymphoblastoid cells is consequently reduced. More than 10% of the EBNA 3C coimmunoprecipitated with J kappa from extracts of non-EBV-infected B lymphoblasts that had been stably converted to EBNA 3C expression. EBNA 3C in nuclear extracts from these cells (or in vitro-translated EBNA 3C) prevented J kappa from interacting with a high-affinity DNA binding site. Under conditions of transient overexpression in B lymphoblasts, EBNA 2 and EBNA 3C associated with J kappa and less EBNA 2 associated with J kappa when EBNA 3C was coexpressed in the same cell. EBNA 3C had no effect on the activity of a -512/+40 LMP1 promoter-CAT reporter construct that has two upstream J kappa sites, but it did inhibit EBNA 2 transactivation of this promoter. These data are compatible with a role for EBNA 3C as a "feedback" down modulator of EBNA 2-mediated transactivation. EBNA 3C could, in theory, also activate transcription by inhibiting the interaction of the J kappa repressor with its cognate DNA. The interaction of two viral transcriptional regulators with the same cell protein may reflect an unusually high level of complexity or stringency in target gene regulation.

Antigens, Viral↗
PubMed1995

Unit-based nursing research.

Sixteen nurses working in a variety of clinical settings met to learn about unit based nursing research. Each had a different perception of the concept but all agreed it was time to validate, via research, what nurses were actually doing to impact patient outcomes. The individual members achieved their goals of learning the steps in the research process and planning a study for their own unit. The mentoring received from the researcher was viewed as a necessary component of accomplishing their projects.

Clinical Nursing Research↗
PubMed1995

Dapsone-induced hemolytic anemia: role of glucose-6-phosphate dehydrogenase in the hemolytic response of rat erythrocytes to N-hydroxydapsone.

Individuals deficient in erythrocytic glucose-6-phosphate dehydrogenase (G6PD) show about a 2-fold increase in sensitivity toward dapsone-induced hemolytic anemia. Rat studies have shown that the hemolytic activity of dapsone resides in its N-hydroxy metabolites; exposure of rat red cells to N-hydroxy-dapsone in vitro followed by readministration to isologous rats results in premature splenic sequestration of the damaged cells. This study examines the ability of the steroid, epiandrosterone, to inhibit rat red cell G6PD and the effect of such inhibition on the susceptibility of rat red cells to N-hydroxydapsone hemolytic activity. Epiandrosterone was found to inhibit rat red cell G6PD uncompetitively and to suppress red cell hexose monophosphate shunt activity by more than 95%. Epiandrosterone suppression of rat red cell G6PD activity resulted in about a 2-fold increase in sensitivity of the rat cells to N-hydroxydapsone hemolytic activity, and a modest but significant increase in depletion of red cell glutathione. In contrast, suppression of rat red cell catalase activity by aminotriazole had no effect on the hemotoxicity of N-hydroxydapsone. Epiandrosterone appears to be a useful tool to explore the mechanism by which G6PD deficiency enhances susceptibility to hemolytic drugs.

Amitrole↗
PubMed1994

Genetic and biochemical evidence that EBNA 2 interaction with a 63-kDa cellular GTG-binding protein is essential for B lymphocyte growth transformation by EBV.

Epstein-Barr virus (EBV) nuclear protein 2 (EBNA 2) is an acidic transcriptional transactivator of virus and cell gene expression and is essential for growth transformation of primary B lymphocytes. EBNA 2 transactivation of response elements (E2REs) can be mediated by interaction with a GTGGGAA-specific DNA-binding factor(s). We now purify the factor by S-Sepharose and EBNA 2 affinity chromatography and identify it as a single 63-kDa protein. The protein is shown to specifically coimmunoprecipitate with EBNA 2 from lymphoblasts transfected with an EBNA 2 FLAG expression vector. Mutation of GTG to TCT in a GTGGGAA motif common to the Cp, LMP2, and LMP1 promoters results in loss of recognition by p63. EBNA 2 amino acids 310-336 are sufficient for p63 binding. The only motif in this 27 amino acid sequence which is common to the EBNA 2 genes of EBV types 1 and 2 is GPPWWPP (I/V) (C/R) DP, which is therefore likely to mediate p63 interaction. Mutation of WW to SS or FF ablates interaction with p63, indicating that both the hydrophobic and aromatic characteristics of WW are essential for its "key" interaction with p63. EBNA 2 with a WW mutated to SS is also unable to marker rescue primary B lymphocyte transforming virus from cells infected with an EBNA 2-deleted virus, while otherwise isogenic wild-type EBNA 2 readily marker rescues transforming virus in parallel experiments. EBNA 2 transactivation through the Cp E2RE is completely abolished by the WW to SS mutation while transactivation of -234 to +40 LMP1 E2RE is only partially affected. These genetic and biochemical experiments support the hypothesis that EBNA 2 WW interaction with a p63 GTGGGAA-binding protein is essential for EBV-mediated cell growth transformation because it specifically associates EBNA 2 with its response elements. This enables the EBNA 2 acidic domain to transcriptionally transactivate specific genes.

Amino Acid Sequence↗
PubMed1994

Glycerol-induced augmentation of sensitivity to endotoxin in rats.

Combined sepsis and rhabdomyolysis result in a mortality rate much higher than that caused by each process alone. An analogous rat model is obtained by simultaneous i.p. administration of a nonlethal dose of lipopolysaccharide (LPS 0.025 mg/100 g) and a nonlethal i.m. injection of glycerol (1 ml/100 g). The aim of this study was to determine the factors contributing to the high mortality rate in this rat model. The factors examined include: Dehydration, plasma volume expansion, 'immunization' to glycerol, induction of LPS tolerance and the effect of free radicals formed in this model. Neither dehydration nor volume expansion affected mortality. 'Immunization' with glycerol was also not effective. In contradistinction, tolerance to LPS achieved by a daily injection with gradual increasing doses of LPS (from 0.05 mg/100 g to 1 mg/100 g) for 6 days reduced the mortality rate by 60% (P < 0.001). Moreover, decreasing free radical activity using the natural antioxidant (NAO) (5 mg/100 g) reduced mortality rates by 50%. A different antioxidant, dimethylthiourea (DMTU) (50 mg/100 g) failed to reduce mortality rates. This study suggests that the synergism between glycerol and LPS is apparently due to an increase in the rats' sensitivity to endotoxin following glycerol injection. However, endotoxin apparently does not enhance sensitivity to glycerol in the rat. The new antioxidant NAO significantly reduced the high mortality rate.

Animals↗
PubMed1993

The interaction between beta-carotene and lipoxygenase in plant and animal systems.

The effect of beta-carotene (BC) on the activity of lipoxygenase (LOX) from plant and animal sources has been examined. Soybean lipoxygenase L-2 activity towards linoleate was inhibited by BC by a maximum of 70% at pH 6.5, whereas L-1 activity was little affected at pH 9.0. Lineweaver-Burk plots indicated that BC inhibited LOX activity by mixed competitive/non-competitive mechanisms. Other hydrophobic compounds also inhibited LOX activity; oleic acid and retinol were competitive inhibitors whereas tocopherol acetate and 5,8,11,14-eicosatetraynoic acid (ETYA) were non-competitive inhibitors. Binding studies with L-2 LOX bound to Sepharose indicated BC-binding and inhibition with the immobilized LOX. Activity of LOX from animal sources was also inhibited by BC both towards linoleate and arachidonate.

Animals↗
PubMed1993

Hepoxilin A3 is the endogenous lipid mediator opposing hypotonic swelling of intact human platelets.

When human blood platelets are exposed to hypotonic medium they swell first but, shortly thereafter, revert toward their original volume in a process termed regulatory volume decrease (RVD). RVD is the result of an enhanced efflux of K+ and Cl- ions and associated water. Platelet RVD is controlled by a short-lived lipoxygenase-derived product (LP). By using a combination of high-performance liquid chromatography, gas chromatography-mass spectrometry, and RVD reconstitution bioassay, we show that LP is identical with hepoxilin A3. In addition we demonstrate that authentic hepoxilin A3 possesses the same biological properties on RVD reconstitution as LP and that the activity of both compounds is amplified through epoxide hydrolase inhibition with 3,3,3-trichloropropene-1,2-oxide. Therefore, we report here that volume expansion causes the formation and release of hepoxilin A3 from intact human platelets and that this hepoxilin plays a major role in volume regulation.

8,11,14-Eicosatrienoic Acid↗
PubMed1993

Lipoxygenase and other enzymes of arachidonic acid metabolism in the brain of chicks affected by nutritional encephalomalacia.

1. Prostaglandin endoperoxide synthetase (PES) and lipoxygenase (Lox) activities were compared in the cerebella and cerebra of vitamin E-sufficient young chicks and in chicks in which nutritional encephalomalacia (NE) was induced by a diet deficient in vitamin E. 2. Eicosanoid production patterns were qualitatively similar in the brains of both groups of chicks, but prostaglandin production was 50-60% less in cerebella of ataxic chicks, compared to control cerebella, while the opposite trend was observed in the cerebellar Lox pathway, as measured by radioimmunoassay of 15-HETE. 3. Cerebellar phospholipase A2 activity was twice that of the cerebrum but was not affected by NE. 4. Purification of Lox activity from the cerebellar homogenates produced a lower yield and enrichment when the starting material was taken from ataxic chicks, compared to the controls. 5. In addition there were qualitative differences in the purified fractions from both groups, as seen by pH optima and kinetics. 6. The results are consistent with the view that the cerebellum has less antioxidant protection than the cerebrum and that its higher phospholipase A2 activity and greater propensity to oxygenate arachidonic acid via the Lox pathway at the expense of the PES pathway may render this region of the brain particularly vulnerable to oxidative damage in NE.

Animal Nutritional Physiological Phenomena↗
PubMed1993

The use of magnetic resonance imaging to track controlled drug release and transport in the brain.

A method has been developed to track controlled drug release and transport in the brain. This method entails the use of a polymeric implant to release, over time, a paramagnetically labelled compound into the brain. Magnetic resonance imaging is used to determine the evolving concentration distribution. This method is well suited to other types of intracranial drug delivery systems as well as to track transport in other organs of the body.

Agar↗
PubMed1993

A strategy for decreasing anxiety of ICU transfer patients and their families.

With the growing number of clients transferred out of the intensive care units (ICUs) following increasingly shorter stays, time constraints have become a barrier to effective teaching. Written information that is readily available to clients helps resolve this problem. A pamphlet (in Spanish and English) was developed to ease the move for patients, families, and critical care and medical nurses from a medical ICU (MICU) to a general floor.

Anxiety↗
PubMed1992

Developmental changes in hepatic basolateral membrane lipid composition and fluidity.

Membrane fluidity and lipid composition influence the activity of a variety of membrane proteins. Decreased rates of hepatic ion clearance are associated with the neonatal period. We postulated that hepatic basolateral membranes derived from suckling animals might be less fluid than those from adult animals. Basolateral membrane vesicles were prepared from the livers of 1-week-old (SBLMV) and adult (ABLMV) rats by a Percoll gradient method. Na+/K(+)-ATPase activities were similar in the two groups. Double bond index, cholesterol and cholesterol/phosphorus ratios were significantly higher in SBLMV compared with ABLMV, while lipid phosphorus and relative percentages of phospholipid subclasses did not differ. Fluorescence anisotropy measured using diphenylhexatriene as well as 2-(9-anthroyloxy)stearate was significantly greater in SBLMV compared with ABLMV, while measurements made with 12-(9-anthroyloxy)stearate were similar in both age groups. Mean excited state lifetimes, lifetime distributions, and rotational correlation times were similar in both groups. These data suggest that hepatic basolateral membranes derived from suckling rats are less fluid than those from adult animals and further suggest that this difference may be due to increased cholesterol in hepatic basolateral membranes derived from suckling animals.

Animals↗
PubMed1992

Lipoxygenase activity in the brain regions of young chicks: isolation and some properties.

1. The lipoxygenase (LOX) oxygenation pathway of arachidonic acid was investigated in the cerebellum and cerebral hemispheres of young chicks. 2. Lipoxygenase products consisted mainly of 15-hydroxyeicosatetraenoic acid (15-HETE), accompanied by the 15-hydroperoxy analog (15-HPETE) and the 5-HETE product. 3. The yield of 15-HETE was 3 times greater in the cerebellar system than in the cerebrum. 4. PLA2 activity of the cerebellum was twice that of the cerebrum. 5. Affinity chromatography revealed 2 brain fractions with LOX activity which were assayed with either linoleic or arachidonic acid as substrate. 6. The fraction eluted with 0.2 M sodium acetate pH 5.0, produced a higher yield and enrichment of LOX activity than the eluate obtained with 0.1 M Tris-HCl buffer (pH 8.0). 7. A considerably higher yield and enrichment of the enzyme was achieved when the starting material was the cerebellum, compared to the cerebrum. 8. The optimal pH for both purified fractions from cerebrum and cerebellum was 6.5, with either linoleic or arachidonic acid as substrate. 9. The cerebral LOX yielded Michaelis-Menten kinetics when linoleic acid was the substrate, while the corresponding plots for the cerebellar enzyme were sigmoidal. 10. Arachidonic acid as substrate produced sigmoidal plots, except at pH 5.0, where Michaelis-Menten kinetics were observed. 11. These results and the elevated activities of PLA2 and 15-LOX could be significant in relation to the special vulnerability of the cerebellum in chick nutritional encephalomalacia.

Animals↗
PubMed1992

Minimizing diagnostic blood loss in critically ill patients.

BACKGROUND: Blood loss from diagnostic procedures in critically ill patients needs to be minimized. Traditionally, when drawing blood from arterial lines, the initial sample used to clear the line has been discarded (open method). Use of a temporary reservoir enables this discard sample to be returned to the patient (closed method). METHODS: Critically ill surgical patients were prospectively randomized to the open or closed method of drawing blood from arterial lines. Blood loss to diagnostic sampling was measured in both groups. RESULTS: A comparison study (n = 1657) of these two methods revealed that blood loss to the patient was significantly decreased (P << .01) using the closed method. Mean blood loss per patient per day was 69 mL in the open group (n = 873) vs 35 mL in the closed group (n = 784). CONCLUSIONS: Use of the closed method when drawing blood from arterial lines results in a significant decrease in blood lost to diagnostic procedures.

Adult↗