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Biomedical subjects

S Liu

Publications and source records attributed to S Liu.

At least 937 records · Page 52Linked to original sources
PubMed1993

Combined scopolamine and morphine treatment of traumatic brain injury in the rat.

Previous studies have indicated that either scopolamine (1.0 mg/kg) or morphine (10.0 mg/kg) administered to rats prior to or soon after moderate fluid percussion traumatic brain injury (TBI) reduces behavioral deficits associated with injury. In this study, a series of experiments examined the effects of a combination of these drugs, as well as each drug individually, on behavioral outcome, brain temperature, and systemic physiological responses to TBI. Experiment I: a single systemic bolus injection of scopolamine (n = 10), morphine (n = 11), scopolamine plus morphine (n = 11), or saline (n = 10) was administered to rats 15 min prior to TBI. Animals were assessed on beam-walking behavioral performance for 5 days after injury. Scopolamine alone or morphine alone significantly reduced (P < 0.05) deficits produced by injury. Treatment with a combination of scopolamine and morphine provided greater protection on beam-walking behavioral measures than either drug alone. Experiment II: morphine raised brain temperature in uninjured rats (n = 5) to a mean of 39.3 degrees C +/- 0.3 by 60 min post-injection. Neither scopolamine (n = 5) nor scopolamine plus morphine (n = 5) altered brain temperature. Experiment III: scopolamine (n = 7) significantly raised heart rate for 5 min after injury. Saline (n = 8), morphine (n = 9) and scopolamine plus morphine (n = 7) significantly lowered heart rate after injury. All four groups had similar hypertensive responses to TBI which peaked at 10 s after injury. The results confirm that pharmacological blockade of muscarinic receptors or stimulation of mu opioid receptors reduces functional deficits associated with TBI.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Chromosome 17p deletions and p53 mutations in renal cell carcinoma.

Studies of the role of tumor suppressor genes in human renal cell carcinoma from our laboratory have suggested the presence of a disease gene(s) on the short arm of chromosome 3. Little is known about the role other tumor suppressor genes may play in this malignancy. Abnormalities of chromosome 17p and, in particular of p53, are common in many human malignancies. In order to evaluate the role of this region in renal cell carcinoma, we performed restriction fragment length polymorphism analyses of chromosome 17 with probes localized to the p53 region. Fourteen of 29 (48%) evaluable cell lines showed loss of heterozygosity at this locus. Northern blot analysis did not detect a p53 transcript in 4 of 27 cell lines tested. In addition, we screened cell lines for p53 mutations using a polymerase chain reaction-single strand conformation polymorphism technique. Cell lines positive for mutations by this technique were then sequenced. Mutations were detected in 11 of 33 (33%) cell lines, including 8 derived from primary tumors and 3 derived from metastatic foci. Six of 9 (67%) patients with loss of heterozygosity demonstrated a mutation in the remaining allele, while only 1 of 8 (13%) without loss of heterozygosity had a mutation. Three of 3 (100%) cell lines derived from metastases had the same mutation as their matched primary cell line. Loss or mutation of p53 did not correlate either with loss of chromosome 3p or with histological subtype. These results suggest that, while the primary disease gene for kidney cancer appears to be on chromosome 3, abnormalities of p53 are common and may be involved in the progression of this malignancy.

Base Sequence↗
PubMed1993

Tyrosine 115 participates both in chemical and physical steps of the catalytic mechanism of a glutathione S-transferase.

The participation of the hydroxyl group of tyrosine 115 in the catalytic mechanism of isoenzyme 3-3 of rat glutathione (GSH) S-transferase is implicated by x-ray crystallographic analysis of a product complex and confirmed by comparison of the catalytic properties of the native enzyme and the Y115F mutant. Tyrosine 115 is located in domain II of the protein (the xenobiotic substrate binding domain) and is the first residue in this domain to be shown to play a direct role in catalysis. The 1.8-A structure of isoenzyme 3-3 in complex with (9S,10S)-9-(S-glutathionyl)-10-hydroxy-9,10-dihydrophenanthrene, one of the diastereomeric products of the reaction of GSH with phenanthrene 9,10-oxide, indicates that the hydroxyl group of Tyr115 is within hydrogen-bonding distance of the 10-hydroxyl group of the bound product and, by implication, is proximal to the oxirane oxygen of the substrate in the Michaelis complex. Site-specific replacement of Tyr115 with phenylalanine has profoundly different effects on catalysis depending on the type of reaction and whether the rate-limiting step in catalysis is a chemical step or a physical step. Stopped flow measurements of the rate constants for product release and viscosity effects on the steady-state kinetics establish that the rate-limiting step in catalysis with phenanthrene 9,10-oxide (kcat = 0.4 s-1) is probably a chemical one, whereas the physical step of product dissociation (koff) is rate-limiting in the reaction of 1-chloro-2,4-dinitrobenzene (kcat = 20 s-1). The Y115F mutant is severely impaired in catalyzing the addition of GSH to phenanthrene 9,10-oxide (kcat = 0.0044 s-1), evidence that the -OH of Tyr115 provides electrophilic assistance in the epoxide ring opening. In contrast, the Y115F mutant is a better catalyst toward 1-chloro-2,4-dinitrobenzene (kcat = 72 s-1) than is the native enzyme. The enhanced rates of product release in the mutant are ascribed to the loss of hydrogen bonds between the -OH of Tyr115 and the side chain -OH and main chain NH of serine 209, interactions that block the channel to the active site or inhibit the segmental motion of the protein.

Amino Acid Sequence↗
PubMed1993

Molecular characterization of the gene encoding the gamma subunit of the human skeletal muscle 1,4-dihydropyridine-sensitive Ca2+ channel (CACNLG), cDNA sequence, gene structure, and chromosomal location.

cDNA clones of the gamma subunit of the skeletal muscle 1,4-dihydropyridine-sensitive voltage-dependent Ca2+ channel were isolated from a human fetal skeletal muscle cDNA library using the rabbit gamma cDNA as a probe. The DNA sequence of the entire human cDNA was determined. Cosmids that contained the human gamma gene were isolated and used to determine the genomic organization of the coding sequences. Four exons were identified, spanning 12.5 kilobases of DNA. Reverse-transcribed polymerase chain reaction analysis detected the gamma transcript in human and mouse skeletal muscle RNAs, but not in RNA from human brain or cardiac muscle or from mouse brain, cardiac muscle, spleen, kidney, liver, or stomach. A polymorphic dinucleotide repeat within the gamma gene was identified. This repeat was used to type a subset of the Centre d'Etude du Polymorphisme Humain families. Linkage analysis indicates that the gamma gene is tightly linked (Z = 12.94, theta = 0.001) to growth hormone at chromosome 17q23, a region that also contains the adult skeletal muscle Na+ channel.

Amino Acid Sequence↗
PubMed1993

Adenosine-5'-carboxaldehyde: a potent inhibitor of S-adenosyl-L-homocysteine hydrolase.

Adenosine-5'-carboxaldehyde (3) and its 4'-epimer (4) were synthesized and shown to be potent type I mechanism-based inhibitors of recombinant rat liver AdoHcy hydrolase with k2/KI values of 16.7 x 10(-3) and 5.5 x 10(-3) nM-1 min-1, respectively. The observation that 3 and 4 are potent inhibitors of AdoHcy hydrolase supports the hypothesis that they function as key intermediates in the mechanism by which the (Z)- and (E)-4',5'-didehydro-5'-deoxy-5'-fluoroadenosines 1 and 2 inactivate this enzyme.

Adenosine↗
PubMed1993

Refined 1.6 A resolution crystal structure of the complex formed between porcine beta-trypsin and MCTI-A, a trypsin inhibitor of the squash family. Detailed comparison with bovine beta-trypsin and its complex.

The crystal structure of the complex formed by porcine beta-trypsin with the MCTI-A inhibitor (Momordica charantia, Linn. Cucurbitaceae) has been determined at 1.6 A resolution using the molecular replacement method. The sequence of MCTI-A was determined by recognizing the electron density, and shows that MCTI-A is a member of the squash family of trypsin inhibitors. We report the first high-resolution structure of porcine beta-trypsin. Detailed comparisons have been made on the overall structure, solvent structure and active-site geometries between this complex and bovine beta-trypsin and its complexes. On the basis of our results, we discuss the interaction patterns between inhibitor and trypsin. Unlike other complex structures formed by bovine trypsin with inhibitors, no out-of-plane distortion around the inhibitor's scissible peptide was observed. The role of the trypsin catalytic triad is also discussed on the basis of this structure.

Animals↗
PubMed1993

Effect of Nai on activity and voltage dependence of the Na/K pump in adult rat cardiac myocytes.

We have measured the voltage dependence of the Na/K pump in isolated adult rat cardiac myocytes using the whole-cell patch-clamp technique. In the presence of 1-2 mM Ba and 0.1 mM Cd and nominally Ca-free, Na/K pump current (Ip) was measured as the change in current due to 1 mM ouabain. Voltage dependence of Ip was measured between -140 and +40 or +60 mV using square voltage-pulse and voltage-ramp protocols, respectively. With 150 mM extracellular Na (Nao) and 5.4 mM extracellular K (Ko), we found that the Na/K pump shows a strong positive voltage dependence between -140 and 0 mV and is voltage independent at positive potentials. Removing Nao reduced the voltage dependence at negative potentials with no effect at positive potentials. When Ko was reduced, a negative slope appeared in the current-voltage (I-V) curve at positive potentials. We have investigated whether Nai (intracellular Na) might also affect the voltage dependence of Ip by varying Na in the patch pipette (Napip) between 20 and 85 mM. We found, as expected, that Ip increased markedly as Napip was raised, saturating at about 70 mM Napip under these conditions. In contrast, while Ip saturated near +20 mV and declined to about 40% of maximum at -120 mV, there was no effect of Napip under these conditions. In contrast, while Ip saturated near +20 mV and declined to about 40% of maximum at -120 mV, there was no effect of Napip on the voltage dependence of Ip. This suggests that neither Nai binding to the Na/K pump nor the conformational changes dependent on Nai binding are voltage dependent. These results are consistent with extracellular ion binding within the field of the membrane but do not rule out the possibility that other steps, such as Na translocation, are also voltage dependent.

Animals↗
PubMed1993

Relationship between tissue calcium content and duodenal ulcer in the rat.

Since the effect of cellular calcium on cell injury has been in question, this study focused on the relationship between tissue calcium content and cysteamine-induced duodenal ulcer. Rats treated with cysteamine showed a high frequency and severity of duodenal ulcer, and the calcium content in the duodenal mucosa was elevated. Furthermore, the level of calcium content in duodenal mucosa was positively associated with the severity of the duodenal lesion. Whereas administration of calcium increased duodenal ulcerative response to cysteamine, verapamil afforded protection against ulceration. We conclude that calcium accumulation in duodenal mucosa is related to duodenal ulceration induced by cysteamine.

Animals↗
PubMed1993

An optimized method for determining cytochrome oxidase activity in brain tissue homogenates.

We have developed a method to accurately and reproducibly determine the total activity of cytochrome oxidase (CO) in rat brain tissue homogenates. Previously, accurate measurements have been difficult to obtain because detergents, which are needed to disrupt membranes and unmask CO, also inhibit the enzyme by solubilizing certain phospholipids required for rapid turnover. We compared various methods of sample preparation, and found that maximal CO activity in homogenates could be obtained using specific concentrations of detergents. The range of optimal detergent concentrations was relatively narrow, as CO activity fell sharply with small deviations from the optimum. Of 5 detergents tested, deoxycholate stimulated CO maximally over the widest range of concentrations. In deoxycholate-treated homogenate samples, the calculated CO turnover number was about 480 s-1, indicating that overall enzyme activity was maximal or near maximal, and therefore that the total content of CO was probably detected. This method was reproducible with large or small samples (e.g., < 1 mg tissue), and should be applicable to studies of neural tissue in general.

Animals↗
PubMed1993

Ecology and evolution of microbial populations for bioremediation.

Bioremediation exploits the genetic diversity and metabolic versatility of microorganisms for the transformation of contaminants into less-harmful end-products, which are then integrated into natural biogeochemical cycles. Understanding the ecology, physiology and evolution of degradative microorganisms is critical for the successful consideration and implementation of bioremediation. This article focuses on the common ecological and evolutionary constraints that influence bioremediation processes.

Bacteria↗
PubMed1993

Potential 99mTc radiopharmaceuticals for renal imaging: tris(N-substituted-3-hydroxy-2-methyl-4-pyridinonato)technetium(IV) cations.

A series of monocationic complexes of N-substituted-3-hydroxy-2-methyl-4-pyridinones labeled with technetium(IV)-99m have been evaluated in vivo as potential radiopharmaceuticals. The pyridinones have different substituents at the ring nitrogen atom: ethyl, i-propyl, i-butyl, benzyl, phenyl, p-methoxyphenyl, 3-butoxypropyl and cyclohexyl. Biodistribution studies of the 99mTc complexes have been carried out in rabbits and mice. High kidney uptake and retention of the radionuclide has been shown in rabbits and mice with the cationic complexes of 3-hydroxy-1-(p-methoxyphenyl)-2-methyl-4-pyridinone and 1-(cyclohexyl)-3-hydroxy-2-methyl-4-pyridinone. These 99mTcL3+ compounds appear to be morphologic renal agents.

Animals↗
PubMed1993

A structural basis for the unequal sensitivity of the major cardiac and liver gap junctions to intracellular acidification: the carboxyl tail length.

The regulation of junctional conductance (Gi) of the major cardiac (connexin43; Cx43) and liver (connexin32; Cx32) gap junction proteins by intracellular hydrogen ion concentration (pH; pHi), as well as well as that of a truncation mutant of Cx43 (M257) with 125 amino acids deleted from the COOH terminus, was characterized in pairs of Xenopus laevis oocytes expressing homologous channels. Oocytes were injected with 40 nl mRNAs (2 micrograms/microliters) encoding the respective proteins; subsequently, cells were stripped, paired, and incubated for 20-24 h. Gj was measured in oocyte pairs using the dual electrode voltage-clamp technique, while pHi was recorded simultaneously in the unstimulated cell by means of a proton-selective microelectrode. Because initial experiments showed that the pH-sensitive microelectrode responded more appropriately to acetate than to CO2 acidification, oocytes expressing Cx32 and wild type and mutant Cx43 were exposed to a sodium acetate saline, which was balanced to various levels of pH using NaOH and HCl. pH was changed in a stepwise manner, and quasi-steady-state Gj -pHi relationships were constructed from data collected at each step after both Gj and pHi had reached their respective asymptotic values. A moderate but significant increase of Gj was observed in Cx43 pairs as pHi decreased from 7.2 to 6.8. In both Cx32 and M257 pairs, Gj increased significantly over a wider pH range (i.e., between 7.2 and 6.3). Further acidification reversibly reduced Gj to zero in all oocyte pairs. Pooled data for the individual connexins obtained during uncoupling were fitted by the Hill equation; apparent 50%-maximum (pK;pKa) values were 6.6 and 6.1 for Cx43 and Cx32, respectively, and Hill coefficients were 4.2 for Cx43 and 6.2 for Cx32. Like Cx32, M257 had a more acidic pKa (6.1) and steeper Hill coefficient (6.0) than wild type Cx43. The pKa and Hill coefficient of M257 were very similar to those of Cx32. These experiments provide the first direct comparison of the effects of acidification on Gj in oocyte pairs expressing Cx43 or Cx32. The results indicate that structural differences in the connexins are the basis for their unequal sensitivity to intracellular acidification in vivo. The data further suggest that a common pH gating mechanism may exist between amino acid residues 1 and 256 in both Cx32 and Cx43. However, the longer carboxyl tail of Cx43 relative to Cx32 or M257 provides additional means to facilitate acidification-induced gating; its presence shifts the pKa from 6.1 (Cx32 and M257) to 6.6 (Cx43) in the conductance of these channels.

Animals↗
PubMed1993

Behavioral protection by moderate hypothermia initiated after experimental traumatic brain injury.

The effects of postinjury hypothermia on behavioral outcome following moderate fluid percussion traumatic brain injury (TBI) were examined. In Experiment I, three groups of rats were examined. The first group was normothermic (37.5 degrees C); and hypothermia (30 degrees C) was initiated 15 min and 30 min postinjury in the second and third groups, respectively. Whole body cooling was achieved by ventral ice pack. Cooling of the brain to 30 degrees C was achieved in 25 min and maintained for 60 min. Brain temperature was measured indirectly by a probe in the temporalis muscle. Behavioral outcome was assessed by beam-balance performance, beam-walking performance, and body weight loss measured daily for 5 days after TBI. Both the normothermic group and the 30-min postinjury hypothermic group exhibited significant (p < 0.05) beam-balance and beam-walking deficits on days 1 through 5 after TBI. In contrast, the 15-min postinjury hypothermic group exhibited significant (p < 0.05) beam-walking deficits only on day 1 after TBI and significant (p < 0.05) beam-balance deficits on days 1, 3, and 4 after TBI. In Experiment II, subcortical brain temperature was compared to temporalis muscle temperature in normothermic (37.5 degrees C) and hypothermic (30 degrees C) rats subjected to TBI. In both groups brain temperature tracked within 0.4 degree C of temporalis muscle temperature. These results are similar to post-TBI excitatory receptor antagonist studies and indicate a therapeutic window for moderate hypothermia of less than 30 min after moderate fluid percussion TBI in the rat.

Animals↗
PubMed1993

Organophosphate sensitizes the human pancreas to acinar cell injury: an ultrastructural study.

Viable pancreas fragments from five human donors were incubated in oxygenated buffered Eagle Medium. The preparation and incubation conditions were based on the method of Scheele and Palade. In Group 1 there was 1-h preincubation with echothiophate (10(-4) M); then, acetylcholine (10(-5) M) was added. After 2 h tissues were prepared for electron microscopy. Acinar injury with vacuole formation was apparent. Many of these changes were observed in fragments incubated only with acetylcholine (10(-5) M) (Group 2) and in incubates with echothiophate only (10(-4) M) (Group 3); only minor changes were seen in controls with Eagle's Medium (Group 4). Large vacuoles were significantly more numerous in Group 1 than in Control Group 4 (p < 0.05). Zymogen granules were depleted in Groups 1, 2, and 3. This depletion was significant in Group 1 when compared with Group 4 (p < 0.02). These results extend previous in vitro results that showed increased amylase release after echothiophate treatment in human pancreas and a left shift in response to acetylcholine.

Acetylcholine↗
PubMed1993

Cellular and molecular aspects of renal repair in acute renal failure.

Experimental data suggest that the recovery of renal function after ischemic or nephrotoxic acute renal failure is due to a replicative repair process dependent on predominantly paracrine release of growth factors. These growth factors promote renal proximal tubule cell proliferation and a differentiation phase dependent on the interaction between tubule cells and basement membrane. These insights identify the molecular basis of renal repair in ischemic and nephrotoxic acute renal failure and may lead to potential therapeutic modalities that accelerate renal repair and lessen the morbidity and mortality associated with these renal disease processes.

Acute Kidney Injury↗