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Biomedical subjects

J Ren

Publications and source records attributed to J Ren.

At least 271 records · Page 15Linked to original sources
PubMed1996

Complexes of HIV-1 reverse transcriptase with inhibitors of the HEPT series reveal conformational changes relevant to the design of potent non-nucleoside inhibitors.

Crystal structures of HIV-1 reverse transcriptase (RT) complexed with a range of chemically diverse non-nucleoside inhibitors (NNIs) have shown a single pocket in which the inhibitors bind and details of the inhibitor-protein interactions. To delineate the structural requirements for an effective inhibitor, we have determined the structures of three closely related NNIs which vary widely in their potencies. Crystal structures of HIV-1 RT complexed with two very potent inhibitors, MKC-442 and TNK-651, at 2.55 angstroms resolution complement our previous analysis of the complex with the less effective inhibitor, HEPT. These structures reveal conformational changes which correlate with changes in potency. We suggest that a major determinant of increased potency in the analogues of HEPT is an improved interaction between residue Tyr181 in the protein and the 6-benzyl ring of the inhibitors which stabilizes the structure of the complex. This arises through a conformational switching of the protein structure triggered by the steric bulk of the 5-substituent of the inhibitor pyrimidine ring.

Antiviral Agents↗
PubMed1996

Correlated flow cytometric analysis of H-ras p21 and DNA ploidy in acute myelogenous leukemia.

The flow cytometric immunoassay was used to study the correlation between the H-ras oncogene product p21 and the DNA ploidy in 30 de novo cases of acute myelogenous leukemia (AML). The results showed that 17 cases were negative for p21 expression and 13 positive for p21. The patients with positive p21 had higher percentage of bone marrow and peripheral blasts and lower peripheral leukocyte count. The expression of p21 had no influence on the therapeutic effect. Before treatment, DNA diploidy occurred in 18 cases including 13 p21 negative ones, and DNA aneuploidy was revealed in 12 cases including 8 p21 positive ones. Patients with positive p21 or having aneuploidy in complete remission were at risk for early relapse. Our results suggest that p21 may be involved in the process of leukemogenesis and progression in AML.

DNA, Neoplasm↗
PubMed1996

Inhibition of progressing primary esophageal peristalsis by pharyngeal water stimulation in humans.

BACKGROUND & AIMS: Sensory impulses initiated from the pharynx exert differing effects on the deglutitive apparatus. They have an inhibitory effect on the lower esophageal sphincter but an excitatory effect on the upper esophageal sphincter. The aim of this study was to systematically investigate the effect of pharyngeal sensory impulses evoked by water stimulation on the progressing esophageal peristalsis. METHODS: Sixteen healthy young volunteers were studied in the supine position. The presence of normal peristalsis was verified. Esophageal peristalsis was recorded 3, 6, 9, 12, 15, and 18 cm above the lower esophageal sphincter. Pharyngeal stimulation was performed by injecting a predetermined threshold volume into the pharynx 2 cm above the upper esophageal sphincter, directed posteriorly. The injections were timed to coincide with the arrival of the peristaltic wave induced by dry swallows at respective recording sites. RESULTS: Injection of the threshold volume (0.5 +/- 0.1 mL) stopped the progression of peristalsis at both the striated and smooth muscle esophagus. Topical pharyngeal anesthesia blocked this inhibitory effect (P < 0.01). CONCLUSIONS: Sensory impulses initiated from the pharynx evoked by water injection inhibit the progression of primary esophageal peristalsis. Although the clinical significance of these findings is not determined, they may explain the mechanism of some of the failed esophageal peristalsis.

Adult↗
PubMed1996

In-vitro activity of levofloxacin against clinical isolates of Legionella spp, its pharmacokinetics in guinea pigs, and use in experimental Legionella pneumophila pneumonia.

The activities of levofloxacin and ofloxacin against 22 clinical legionella isolates was determined by microbroth dilution susceptibility testing. Growth inhibition of two Legionella pneumophila strains grown in guinea pig alveolar macrophages by levofloxacin, ofloxacin, or erythromycin was also determined. The drug concentrations required to inhibit 90% of strains tested was 0.032 mg/L for levofloxacin or ofloxacin, and was 0.016 mg/L for ciprofloxacin. BYE alpha broth significantly inhibited the activities of all three drugs tested, as judged by the susceptibility of control Escherichia coli strains. Levofloxacin (0.25 mg/L) reduced bacterial counts of two L. pneumophila strains grown in guinea pig alveolar macrophages by 1 log10, but regrowth occurred over a 3 day period; levofloxacin (1 mg/L) reduced bacterial counts by 2-3 log10 cfu/mL. Levofloxacin was significantly more active than erythromycin, and as active as ofloxacin or ciprofloxacin in this assay. Pharmacokinetic and therapy studies of levofloxacin and ofloxacin were performed in guinea pigs with L. pneumophila pneumonia. For the pharmacokinetic study, levofloxacin was given (10 mg/kg) by the intraperitoneal route to infected guinea pigs; mean peak plasma and lung concentrations were 3.4 mg/L and 1.4 micrograms/g, respectively, at 0.5 h and 2.6 mg/L and 0.6 micrograms/g at 1 h. The terminal half-life phase of elimination from plasma and lung was c. 1 h. All 15 infected guinea pigs treated with levofloxacin (10 mg/kg/day given ip once daily) for 5 days survived for 9 days after antimicrobial therapy, as did all 14 guinea pigs treated with the same dose of ofloxacin. None of 13 animals treated with saline survived. Levofloxacin is effective against L. pneumophila in vitro and in a guinea pig model of legionnaire's disease. Levofloxacin should be evaluated as a treatment of human legionnaires' disease.

Animals↗
PubMed1996

Activity of trovafloxacin (CP-99,219) against Legionella isolates: in vitro activity, intracellular accumulation and killing in macrophages, and pharmacokinetics and treatment of guinea pigs with L. pneumophila pneumonia.

The activity of trovafloxacin against 22 clinical Legionella isolates was determined by broth microdilution susceptibility testing. The trovafloxacin concentration required to inhibit 90% of strains tested was < or = 0.004 micrograms/ml, in contrast to 0.032 micrograms/ml for ofloxacin. In guinea pig alveolar macrophages, trovafloxacin achieved intracellular levels up to 28-fold over the extracellular concentration, which was similar to the levels obtained with erythromycin. Trovafloxacin (0.25 micrograms/ml) reduced bacterial counts of two L. pneumophila strains grown in guinea pig alveolar macrophages by > 2 log10 CFU/ml, without regrowth, under drug-free conditions over a 3-day period; trovafloxacin was significantly more active than ofloxacin or erythromycin (0.25 to 1 microgram/ml) in this assay. Single-dose (10 mg of prodrug CP-116,517-27 per kg of body weight given intraperitoneally [i.p.], equivalent to 7.5 mg of trovafloxacin per kg) pharmacokinetic studies performed in guinea pigs with L. pneumophila pneumonia revealed peak serum and lung trovafloxacin levels to be 3.8 micrograms/ml and 5.0 micrograms/g, respectively, at 0.5 h and 4.2 micrograms/ml and 2.9 micrograms/g, respectively, at 1 h. Administration of a lower prodrug dose (1.4 mg of trovafloxacin equivalent per kg i.p.) gave levels in lung and serum of 0.4 microgram/g and 0.4 microgram/ml, respectively, 1 h after drug administration. The terminal half-lives of elimination from serum and lung were 0.8 and 1.1 h, respectively. All 15 infected guinea pigs treated for 5 days with CP-116,517-27 once daily (10 mg/kg/day i.p., equivalent to 7.5 mg of trovafloxacin per kg/day) survived for 10 days after antimicrobial therapy, as did all 15 guinea pigs treated with ofloxacin once daily (10 mg/kg/day i.p.) for 5 days. None of 13 animals treated with saline survived. In a second experiment with animals, trovafloxacin (1.4 mg/kg/day i.p. for 5 days) protected all 16 guinea pigs from death, whereas all 15 animals treated with saline died. Trovafloxacin is an effective antimicrobial agent against Legionella in vitro and in vivo, with the ability to concentrate in macrophages and kill intracellular organisms.

Animals↗
PubMed1996

Kinematic and dynamic characteristics of solid pellet movement during the pharyngeal phase of swallowing.

Information on solid particle movement through the pharynx, in addition to its physiologic importance, has relevance to the swallowing of medications in pill form. The purpose of this study was to determine the kinematics of a solid particle during the oral-pharyngeal phase of swallowing. We used a concurrent manometric-videofluoroscopic technique and identified two distinct zones of increasing bolus velocity, one at the tongue base and the other at the pharyngo-upper esophageal sphincter (UES) region. Velocity decreased significantly (p < .05), to 9.0 +/- 1.0 cm/s, while the bolus traversed the area located between the tip of the horizontal epiglottis and the pharyngeal wall. The velocity of the liquid barium bolus head was similar to that of the solid barium pellet. The average bolus tail velocity was relatively constant. The acceleration of the barium pellet was temporally associated with development of an incrementally decreasing pressure distal to the location of the pellet in the hypopharynx and across the UES. In conclusion, the kinematics of a solid particle are similar to those of the head of a liquid bolus, but both are different from bolus tail kinematics. During the pharyngeal phase of swallowing, the area located ahead of the bolus exhibits an incrementally decreasing pressure, caudally. This may facilitate bolus transport and contribute to airway protection.

Adult↗
PubMed1996

Troglitazone attenuates high-glucose-induced abnormalities in relaxation and intracellular calcium in rat ventricular myocytes.

Diabetes is associated with impaired cardiac diastolic dysfunction. Isolated ventricular myocytes from diabetic animals demonstrate impaired relaxation concomitant with prolonged intracellular Ca2+ transients. We have recently shown that maintaining normal adult rat ventricular myocytes in a "diabetic-like" culture medium (low insulin and high glucose) produces abnormalities in excitation-contraction coupling similar to in vivo diabetes. Troglitazone (TRO), a novel insulin-sensitizing agent, significantly lowers blood pressure and modestly increases cardiac output in vivo, but its direct impact on cardiac function is unknown. To determine whether TRO could prevent high-glucose-induced dysfunction, normal myocytes were maintained in culture for 1-2 days in either normal medium containing 5 mmol/l glucose or high-glucose medium containing 25 mmol/l glucose. TRO (5 micromol/l) was added to both normal and high-glucose media. Mechanical properties were evaluated using a high-resolution video-edge detection system, and Ca2+ transients were recorded in fura-2-loaded myocytes. Relaxation from peak contraction was significantly longer in myocytes cultured in high glucose. Treating cells with TRO either attenuated or prevented the high-glucose effects, without changing the mechanical properties of myocytes cultured in normal medium. TRO also prevented the abnormally slow rates of Ca2+ transient decay induced by high glucose. Collectively, these data demonstrate that TRO can protect against the high-glucose-induced relaxation defects, perhaps through changes in intracellular Ca2+ handling. If TRO has both vasodilatory actions and beneficial cardiac properties (e.g., improvement of diastolic function) in the presence of hyperglycemia, this antidiabetic agent may prove to have significant salutary cardiovascular effects in type II diabetes.

Animals↗
PubMed1996

Induction of immunotolerance in rats by intratesticular administration of an eicosapeptide of bovine S-antigen.

Immunization of albino LEW rats with a retinal soluble antigen (S-antigen) induces experimental autoimmune uveoretinitis (EAU) which shows clinical features resembling those of human uveitis. Several uveitogenic epitopes have been identified in the antigen. This study reports that an intratesticular injection of low doses of a uveitogenic eicosapeptide (P343-362) of S-antigen prior to immunization with the same peptide prevented the onset of EAU by inducing systemic tolerance, designated orchidic tolerance. Splenic lymphocytes of both CD4+ and CD8+ subsets from tolerized rats transferred orchidic tolerance to syngeneic recipients and protected them from subsequent EAU induction. Orchidic tolerance elicited by low antigen dosage was mediated, in part, by active suppression due to suppressor or regulatory cells. At high antigen doses, however, regulatory activity was reduced possibly due to the induction of anergy in regulatory cells, and EAU severity increased. The CD4+ regulatory T cells from tolerized rats showed enhanced expression of IL-4 mRNA compared with CD4+ cells from control rats. Increased immunoreactivity for IL-4, IL-10 and TGF-beta was observed in the spleen and lymph nodes of tolerized animals. The results suggest that orchidic tolerance induced by low doses of P343-362 is mediated in part by CD4+ regulatory cells secreting Th2 cytokines.

Adoptive Transfer↗
PubMed1996

The actions of prostaglandin E2 on potassium currents in rat tail artery vascular smooth muscle cells: regulation by protein kinase A and protein kinase C.

In vascular smooth muscle cells (VSMCs) of rat tail artery, prostaglandin E2 (PGE2) inhibited a voltage-dependent, delayed rectifier K channel current (Ik). The inhibition was concentration-dependent, via a receptor-mediated mechanism involving the activation of G protein(s) (Ren et al., 1995). In this study, we show that the PGE2-induced inhibition of Ik was mediated by activation of protein kinase A (PKA) and possibly protein kinase C (PKC). Pretreatment of the cells with cyclic adenosine 3',5'-monophosphothioate Rp-isomer (Rp-cAMPs), an inhibitor of adenosine 3', 5'-cAMP-dependent protein kinase (PKA), almost completely abolished the PGE2-induced inhibition. Forskolin, dibutyryl cAMP (Db-cAMP) and cyclic adenosine 3',5'cyclic monophosphothioate Sp-isomer (Sp-cAMPs), activators of adenylate cyclase and PKA, mimicked the effect of PGE2 on Ik. Phosphodiesterase inhibition by 3-isobutyl-1-methylxanthine did not alter the PGE2-induced inhibition of Ik. Moreover, we also found that phorbol myristate acetate (PMA), a PKC activator, significantly suppressed Ik. Both the kinase inhibitor staurosporine and down-regulation of PKC by prolonged exposure of the cells to PMA blocked the PGE2-induced inhibition of Ik, but had no effects on the forskolin, Db-cAMP or SpcAMP-induced effect on Ik. Pretreatment of the cells with Rp-cAMPs only partially diminished the degree of Ik inhibition evoked by PMA. Assay of cAMP content indicated that both PGE2 and PMA induced cAMP accumulation. These results strongly suggest that the modulation of Ik by PGE2 in rat tail artery VSMCs involves signal transduction through both PKA and PKC activation. The activation of PKC may potentiate the cAMP-PKA stimulation, whereas the cAMP-PKA cascade did not seem to affect the PKC pathway. These observations suggest that "cross talk" between the two second-messenger systems is involved in the mechanisms that mediate the effect of PGE2.

1-Methyl-3-isobutylxanthine↗
PubMed1996

Coordination of deglutitive vocal cord closure and oral-pharyngeal swallowing events in the elderly.

OBJECTIVE: The temporal relationship between deglutitive glottic closure, oropharyngeal bolus transit and other biomechanical swallowing events was studied in 10 healthy elderly subjects by concurrent transnasal videolaryngoscopy, videofluoroscopy, intraluminal pharyngeal manometry and submental electromyography. The results were compared with those of a similar study in a group of young healthy volunteers published previously. RESULTS: Overall, the coordination of various deglutitive biomechanical events in the elderly group was found to be similar to that in the young. However, subtle differences were documented. Contrary to historical young controls, where the onset of the bolus movement from the mouth toward the pharynx invariably occurred after the vocal cords reached their maximum adduction, in the elderly this relationship was variable. In all swallows of two subjects it occurred 0.41 +/- 0.2 s after, and in another two it occurred 0.08 +/- 0.06 s before the maximal vocal cord adduction. Six subjects showed interchanging patterns. Except for the above-mentioned changes, the onset of vocal cord adduction in the elderly also preceded all other studied events, similar to previously published data in the young. In the elderly, the total duration between onset of vocal cord adduction and their complete reopening (2.47 +/- 0.1 s) was similar to that in the young. CONCLUSION: Although subtle alterations occur in the coordination of deglutitive vocal cord closure and oropharyngeal bolus transit in some elderly people, the overall coordination between airway protection and the transit aspect of the oral-pharyngeal phase of swallowing is preserved in the elderly.

Aged↗
PubMed1996

Experimental autoimmune uveoretinitis induced by the gamma-subunit of cyclic guanosine monophosphate phosphodiesterase in rats.

PURPOSE: To investigate the capacity of the recombinant gamma-subunit (P gamma) of cyclic guanosine monophosphate phosphodiesterase to induce experimental autoimmune uveoretinitis in Lewis rats. METHODS: Bovine P gamma was expressed in Escherichia coli cells and purified by fast protein liquid chromatography. Lewis rats were immunized by a single footpad injection of P gamma emulsified in complete Freund's adjuvant. Clinical and histopathologic changes in the eye and pineal gland were examined. Lymphocytes were prepared from the lymph nodes of rats with uveitis and transferred by intraperitoneal injection to naive recipient rats. RESULTS: Immunization of rats with P gamma induced panuveitis and pinealitis with clinical and histopathologic changes similar to those induced by S-antigen. Lymphocytes from the lymph nodes of diseased rats transferred uveitis to naive recipients. CONCLUSIONS: P gamma, a retina-specific protein of molecular weight less than 10,000 kDa, is capable of inducing uveoretinitis in Lewis rats. The disease can be transferred adoptively to naive rats by injection of lymphocytes from donor rats with experimental autoimmune uveoretinitis. Inflammation of the pineal gland of immunized rats suggests that P gamma is not only localized to the retina but also to the pineal gland.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1996

[The turn-over flap of the frontalis muscle used for eye-socket depression with contraction of the conjunctival capsule].

We were used to repair the eye-socket depression and contraction after eyeball loss with fat, dermis or rib cartilage implantation. This "stuffing method" has some disadvantages, including absorption and exposure of the implant. In recent years the authors have used a turn-over flap of the frontalis muscle to treat eye-socket depression with contraction of the conjunctival capsule. Satisfactory results have been found at postoperative follow-up.

Adolescent↗
PubMed1996

[ATP-activated single ion channel and its properties in cerebral cortical neurons of neonatal rat].

In the present study, single channel currents were recorded from single neurons dissociated from the cerebral cortex of neonatal SD rat by the cell-attached and inside- out modes of patch clamp technique. ATP-activated channels with a conductance of 32 pS were often recorded and permeable to Na+, K+ and Cs+, but not to Cl-. Most of open time distribution histograms were fitted by two exponential components and few by one, whereas closed time distribution histograms were fitted by two exponential components. Both mean open time and open probability were independent of membrane potential, but the latter increased with increasing ATP concentration. The channel activity was recorded only in the presence of pipette solution containing ATP, and its conductance and open probability were not affected by ACh N-receptor antagonist hexamethonium and mecamylamine. It is suggested that ATP-activated ion channels probably exist in the cerebral cortical neurons of neonatal rat.

Adenosine Triphosphate↗
PubMed1995

Mechanisms of proton-induced stimulation of CGRP release from rat antrum.

Mechanisms of acid-evoked CGRP release from gastric afferent nerves were investigated in rat antral mucosal/submucosal tissues. Low pH (pH 4.0, 5.0 and 6.0) stimulated antral CGRP release significantly and dose-dependently from rat antral fragments. Removal of extracellular calcium from the incubation medium resulted in significant inhibition (59%, P < 0.001) of acid (pH 4.0)-stimulated CGRP release. Conotoxin (1 x 10(-7) M), the selective blocker of N-type calcium channels, also significantly inhibited proton (pH 4.0)-induced CGRP release to values that were 74% below net stimulated levels. Neither nifedipine (1 x 10(-6) M), the L-type Ca(2+)-channel antagonist, nor indomethacin (1 x 10(-5) M), inhibitor of prostaglandin synthesis, altered acid-induced CGRP release. In contrast, ruthenium red (1 x 10(-5) M), capsaicin antagonist, almost completely prevented acid (pH 4.0)-stimulated CGRP release. Capsazepine (1 x 10(-4) M), a specific capsaicin receptor antagonist, also completely abolished acid-induced CGRP release. In conclusion, the results of these studies indicate that hydrogen ions are capable of evoking CGRP release from peripheral sensory neurons in rat antral mucosal/submucosal tissues. Proton-evoked CGRP release requires extracellular calcium and involves N-type calcium channels. Furthermore, acid appears to exert a capsaicin-like effect to evoke sensory neuropeptide release that is sensitive to capsazepine and ruthenium red. These data suggest that proton-induced antral CGRP release represents a direct action of hydrogen ions on mucosal/submucosal sensory dendritic nerve endings to effect local release of neuropeptide.

Animals↗
PubMed1995

The structure of HIV-1 reverse transcriptase complexed with 9-chloro-TIBO: lessons for inhibitor design.

BACKGROUND: HIV reverse transcriptase (RT) is a key target of anti-AIDS therapies. Structural studies of HIV-1 RT, unliganded and complexed with different non-nucleoside inhibitors (NNIs), have pointed to a common mode of binding and inactivation through distortion of the polymerase catalytic site by NNIs containing two hinged rings. The mode of binding of the TIBO family of inhibitors is of interest because these compounds do not fit the two-hinged-ring model. RESULTS: The structure of HIV-1 RT complexed with 9-chloro-TIBO (R82913) has been determined at 2.6 A resolution. As reported for the lower resolution analysis of another TIBO compound, this inhibitor binds at the same site as other NNIs, but our higher resolution study reveals the Cl-TIBO is distorted from the conformation seen in crystals of the inhibitor alone. This allows Cl-TIBO to mimic the binding of NNIs containing two hinged rings. Inhibitor-protein interactions are again predominantly hydrophobic and the protein conformation corresponds to that seen in complexes with other tight-binding NNIs. CONCLUSIONS: Although Cl-TIBO is chemically very different from other NNIs, it achieves remarkable spatial equivalence and shape complementarity with other NNIs on binding to RT. Comparison of the different RT-NNI complexes suggests modifications to the TIBO group of inhibitors which might enhance their binding and hence, potentially, their therapeutic efficacy.

Benzodiazepines↗
PubMed1995

Purification and properties of a Golgi-derived (alpha 1,2)-mannosidase-I from baculovirus-infected lepidopteran insect cells (IPLB-SF21AE) with preferential activity toward mannose6-N-acetylglucosamine2.

Because the availability and subcellular distribution of processing mannosidases in cells play such powerful roles in determining ultimate structures of glycoconjugates, we desired to identify, characterize, and investigate possible regulation of mannosidases in infected and noninfected lepidopteran insect cells. Since our previous observations that a mannosidase activity that converted Man6GlcNAc2 to Man5GlcNAc2 was enhanced in virus-infected cells, thus providing the necessary intermediate for further processing to complex-type oligosaccharides, we attempted purification of this enzyme. A mannosidase was isolated and purified from membranes, operationally defined as Golgi, of recombinant baculovirus-infected Spodoptera frugiperda (IPLB-SF-21AE) cells. The molecular mass of this protein was approximately 63 kDa. Assays performed by measuring the conversion of NaB3H4-reduced Man6GlcNAc2-ol to Man5GlcNAc-[3H]GlcNAc2-ol demonstrated that the mannosidase activity was dependent on the presence of divalent cations, which was optimal for Ca2+ at pH 6.0. Inclusion of 1-deoxymannojirimycin resulted in 50% inhibition at a concentration of 20 microM, whereas swainsonine did not show such inhibition. No activity was observed with p-nitrophenyl alpha-D-mannoside (4 mM) as a substrate. The preferred reduced oligosaccharide substrate was Man6GlcNAc2-ol, with lower activities obtained with Man9GlcNAc2-ol, Man8GlcNAc2-ol, and Man7GlcNAc2-ol. With Man6GlcNAc2-ol as substrate, products smaller than reduced Man5GlcNAc2-ol were not observed. Mannose was also liberated from the glycoprotein, ovalbumin. These properties are consistent with an enzyme classification as a type I (alpha 1,2)-Man6-mannosidase.

Animals↗
PubMed1995

Scanning Kelvin Microscope: a new method for surface investigations.

Based on the well known Kelvin probe for work function measurements a new microstructure analysis system - the Scanning Kelvin Microscope - has been developed. It allows to measure simultaneously with high lateral resolution the distribution of the contact potential difference (CPD) between a conductive sample and a reference probe together with the topographical structure of the sample surface. The measurement is contact free and non-destructive and can be carried out in natural environments. At present the lateral resolution of the measurement approaches 5 microm. The results can be displayed on a computer in three dimensional colour pictures.

Journal Article↗