Biomedical subjects

R Ho

Publications and source records attributed to R Ho.

At least 37 records · Page 2Linked to original sources

PubMed1996

Chaperonins GroEL and GroES: views from atomic force microscopy.

The Escherichia coli chaperonins, GroEL and GroES, as well as their complexes in the presence of a nonhydrolyzable nucleotide AMP-PNP, have been imaged with the atomic force microscope (AFM). We demonstrate that both GroEL and GroES that have been adsorbed to a mica surface can be resolved directly by the AFM in aqueous solution at room temperature. However, with glutaraldehyde fixation of already adsorbed molecules, the resolution of both GroEL and GroES was further improved, as all seven subunits were well resolved without any image processing. We also found that chemical fixation was necessary for the contact mode AFM to image GroEL/ES complexes, and in the AFM images. GroEL with GroES bound can be clearly distinguished from those without. The GroEL/ES complex was about 5 nm higher than GroEL alone, indicating a 2 nm upward movement of the apical domains of GroEL. Using a slightly larger probe force, unfixed GroEL could be dissected: the upper heptamer was removed to expose the contact surface of the two heptamers. These results clearly demonstrate the usefulness of cross-linking agents for the determination of molecular structures with the AFM. They also pave the way for using the AFM to study the structural basis for the function of GroE system and other molecular chaperones.

Adenylyl Imidodiphosphate↗

PubMed1996

Differential gene expression of transforming growth factors alpha and beta, epidermal growth factor, keratinocyte growth factor, and their receptors in fetal and adult human prostatic tissues and cancer cell lines.

OBJECTIVES: Recent studies have shown that growth factors may play a role in the etiology of benign prostatic hyperplasia (BPH) and prostatic carcinoma. Several growth factors have been reported to be expressed by prostatic tissues, but these growth factors have never been examined in human fetal prostate and compared with adult prostates and cancer cell lines. The present study was designed to investigate the messenger ribonucleic acid (mRNA) expression of transforming growth factor (TGF)-alpha, TGF-beta 1, TGF-beta 2, TGF- beta 3, keratinocyte growth factor (KGF), epidermal growth factor (EGF), EGF receptor (EGF-R), and KGF receptor (KGF-R) in human fetal and adult prostatic tissues and cancer cell lines by reverse-transcriptase-polymerase chain reaction-(RT-PCR) using specific oligonucleotide primers. METHODS: Total RNA was extracted from human fetal and adult prostates (BPH tissues) and cancer cell lines. The gene expression of these growth factors and their receptors was determined by RT-PCR using specific oligonucleotide primers. RESULTS: The results of these experiments suggest that: (1) human fetal prostate expressed mRNA transcripts for TGF-alpha, TGF-beta 1, TGF-beta 2, TGF-beta 3, and EGF. However, KGF, KGF-R, and EGF-R mRNA were not expressed by human fetal prostate; (2) human adult prostate (BPH tissues) showed mRNA transcripts for all growth factors and their receptors except KGF-R; (3) human BPH-1 cell lines expressed mRNA transcripts for TGF-alpha, TGF-beta 1, TGF-beta 2, TGF-beta 3, EGF, and KGF-R, but not for EGF-R and KGF growth factors; (4) human primary prostate cancer cell line (ND-1) showed mRNA transcripts for all growth factors except EGF and KGF; and (5) human prostate cancer cell lines (LNCaP, DU-145, PC-3) expressed mRNA transcripts for all growth factors except KGF, which was absent in all cell lines. However, KGF-R mRNA was absent in the PC-3 prostate cancer cell line. CONCLUSIONS: These results suggest that the differential gene expression for various growth factors and their receptors in human fetal and adult prostatic tissues and cancer cell lines may be important in understanding the role of these factors in the pathophysiology of prostatic diseases.

Adult↗

PubMed1995

Touch-evoked agitation produced by spinally administered phospholipid emulsion and liposomes in rats. Structure-activity relation.

BACKGROUND: Phospholipid-based liposomes can alter the kinetics of spinally administered agents. We have observed that spinal delivery of these preparations results in an unexpected touch-evoked agitation, a state in which light touch evokes vocalization by the rat. In the current study we characterized this agitated state induced by various liposome preparations. METHODS: Rats prepared with lumbar intrathecal catheters received a variety of phospholipids delivered spinally as emulsions or as liposomes. Before and after injection, the animal's hot-plate latency (52.5 degrees C surface), spontaneous mobility, and spontaneous and evoked pain behavior were assessed. RESULTS: Spinal delivery of L-alpha-phosphatidylcholine of egg yolk (L-EPC) and the phospholipase hydrolysis product (lyso-L-alpha-phosphatidylcholine [lyso-L-EPC]) produced dose-dependent touch-evoked agitation; the order of potency and rapidity of onset was lyso-L-EPC > L-EPC, with no difference in activity whether administered as an emulsion or as a liposome preparation. Examination of the activity of a series of pure phospholipids revealed the ordering of touch-evoked agitation potency (where PC = phosphatidylcholine) to be L-monopalmitoyl-PC (lyso product of L-dipalmitoyl-PC) > L-dipalmitoyl-PC > L-distearoyl-PC, L-dioleoyl-PC >> L-dilauroyl-PC > L-dimyristoyl-PC; D-dipalmitoyl-PC = 0. The effect of unsaturation on touch-evoked agitation cannot be predicted because dioleoyl-PC and dioleoyl phosphatidylglycerol produced touch-evoked agitation but dipalmitoleoyl-PC did not. Substitution of glycerol for choline as the head group had no influence on touch-evoked agitation. Spinal treatment with an inhibitor of phospholipase (mepacrine) or a cyclooxygenase (ketorolac) blocked the touch-evoked agitation of L-EPC but not that of lyso-L-EPC. CONCLUSIONS: These results emphasize that certain L-isomeric phospholipids with their gel-transition temperatures near body temperature can produce prominent touch-evoked agitation after spinal delivery, an effect likely mediated by a phospholipase hydrolysis product. This touch-evoked agitation, which is consistent with neurotoxicity reported in the early literature on lysophospholipids, suggests that the choice of lipids for the formulation of liposomes intended for spinal drug delivery should be carefully considered.

Animals↗

PubMed1994

A high-sensitivity CCD system for parallel electron energy-loss spectroscopy (CCD for EELS).

A cooled frame transfer CCD camera system was developed and tested as a parallel detector in an electron energy-loss spectrometer mounted on a transmission electron microscope. The use of a shutterless camera with a frame transfer CCD collected virtually 100% of the photon signal with a reasonably fast acquisition time. The system detective quantum efficiency was over 90% under normal experimental conditions. Because of the low channel to channel gain variations in the CCD, the signal-to-noise ratio and the detection limit were substantially better than that obtained with a silicon intensified target (SIT) camera, and direct fitting to the standard data was feasible. Quantitation at the phosphorus L edge generated from a phosphoprotein, phosvitin, showed that, under identical experimental conditions, direct fitting of spectra obtained with this CCD system gave better sensitivity than that given by the SIT camera system. Because of its larger pixel charge well, the CCD system can also operate at a much higher beam current, resulting in a significant reduction in the time required for elemental mapping at a given sensitivity.

Image Processing, Computer-Assisted↗

PubMed1993

Thickness determination of biological thin sections by multiple-least-squares fitting of the carbon K-edge in the electron energy-loss spectrum.

We show that the local thickness of biological thin sections can be measured by fitting the electron energy-loss spectrum of the plurally scattered carbon K-edge, and that such measurements are consistent with the results obtained by a conventional method based on the relationship t/lambda alpha ln(It/I0). The new method allows the simultaneous measurement and/or mapping of both local thickness and core-level excitations (e.g. Ca L-edge) in this energy range (280-400 eV) without requiring the measurement of the zero-loss and the valence/plasmon excitations.

Animals↗

PubMed1992

Euthanasia and abortion: personality correlates for the decision to terminate life.

The study addresses three research questions: (a) How well do level of conservatism and religiosity predict attitudes toward passive and active euthanasia? (b) What is the relationship between attitudes toward abortion and attitudes toward euthanasia? (c) How do personality variables relate differentially to these attitudes? Subjects were 168 Australian adults (68 men, 100 women). Initial analysis indicated no sex differences in attitudes toward the study's criterion variables. Descriptive statistics indicated general support for active and passive euthanasia and for abortion, although the majority of the respondents were found to be more accepting of passive than of active euthanasia. Multiple regression analyses indicated that level of conservatism was the most consistent predictor of attitudes toward euthanasia and abortion, whereas religiosity was found significantly to predict attitudes toward abortion only.

Abortion, Induced↗

PubMed1990

Stimulatory role of substance P on gonadotropin release in ovariectomized rats.

Substance P (SP) has been shown to be present in the hypothalamus and anterior pituitary. To evaluate a possible physiological role of endogenous SP in the control of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) release, specific antiserum against SP (anti-SP) was injected intraventricularly (3 microliters into the third ventricle) or intravenously (50 or 200 microliters) into conscious, ovariectomized (OVX) rats. Third ventricular injection of the antiserum induced a significant decrease in both plasma LH and FSH levels when compared to values in control animals injected with normal rabbit serum (p less than 0.01 and p less than 0.025, respectively). The effect was observed within 10 mi and levels remained suppressed for 60 min. In contrast, intravenous injection of large doses of anti-SP had no effect on the release of both hormones. In order to confirm the stimulatory effect of SP itself, synthetic SP was injected intravenously and intraventricularly into estrogen-primed (E-primed), OVX rats. Synthetic SP dramatically stimulated LH release, but not FSH release when injected either intravenously or intraventricularly at doses of 10 and 50 micrograms (p less than 0.001, p less than 0.005 vs. control, respectively). To investigate any direct action of SP on gonadotropin release from the anterior pituitary gland, synthetic SP was incubated with dispersed anterior pituitary cells harvested from E-primed OVX rats. SP did not affect the release of gonadotropins in vitro. These results indicate that endogenous hypothalamic SP exerts a tonic stimulatory hypothalamic control of basal gonadotropin release in OVX rats.

Animals↗

PubMed1989

Cerebrovascular disease and functional outcome after coronary artery bypass surgery.

A series of patients undergoing coronary artery bypass surgery was studied prospectively to see if angiographic evidence of cerebrovascular disease proved predictive of the incidence of neuropsychological deficit 8 days or 8 weeks after surgery. In 47 patients, intravenous digital subtraction angiography was carried out preoperatively to assess the presence and severity of atheromatous changes in the carotid arteries; 51% had evidence of vessel wall disease and 17% had stenosis of at least one carotid artery in the neck, although only one patient had severe narrowing. Overall, 77% of these 47 patients showed a neuropsychological deficit as defined by a significantly reduced score in at least two of 10 tests administered 8 days after surgery. Eight weeks after surgery 36% still showed a deficit. The incidence of neuropsychological deficit was not significantly greater among those patients with angiographically visible carotid artery disease. The mechanism of surgery-related cognitive impairment is briefly discussed in the light of these findings.

Brain↗

PubMed1988

Peripheral neuropathy complicating pancreatitis and major pancreatic surgery.

Four cases of a polyneuropathy associated with pancreatitis and pancreatic pseudocyst formation are reported. Electrophysiological investigation showed the peripheral neuropathy to be predominantly axonal in type. These patients were all seriously ill and many factors may have been involved in the pathogenesis of their neuropathy. They had all received parenteral nutrition and multiple drug therapy including metronidazole, and all had severe sepsis. There was evidence that insufficient vitamin replacement had been given during total parenteral nutrition. It was not possible to decide whether the polyneuropathy resulted from the summation of these factors, is similar to what has been called the polyneuropathy of the critically ill, or is a new association with pancreatic disease.

Acute Disease↗

PubMed1987

Development of brainstem and cerebellar projections to the diencephalon with notes on thalamocortical projections: studies in the North American opossum.

The North American opossum is born in a very immature state, 12 days after conception, and climbs into an external pouch where it remains attached to a nipple for an extended period of time. We have taken advantage of the opossum's embryology to study the development of brainstem and cerebellar projections to the diencephalon as well as the timing of diencephalic projections to somatosensory motor areas of neocortex. The techniques employed included immunocytochemistry for serotonin, the retrograde and orthograde transport of wheat germ agglutinin conjugated to horseradish peroxidase, and the selective impregnation of degenerating axons. Our results suggest that serotoninergic axons, presumably from the dorsal raphe and superior central nuclei, are present in the diencephalon at birth. Axons from the bulbar reticular formation, the vestibular complex, the trigeminal sensory nuclei, and the dorsal column nuclei reach at least mesencephalic (and probably diencephalic) levels by postnatal day (PND) 3, whereas those from the cerebellar nuclei may not grow into comparable levels until PND 5. The dorsal column and cerebellar nuclei innervate the ventral nuclei of the thalamus by estimated postnatal day (EPND) 17 and all of the diencephalic nuclei supplied in the adult animal by EPND 26. Diencephalic axons enter ventrolateral (face) areas of presumptive somatosensory motor cortex by PND 12, but do not reach dorsomedial (limb) regions until EPND 21. At both ages, diencephalic axons are limited to the cortical subplate and marginal zone; they do not innervate an identifiable internal granular layer until considerably later. Our results suggest that axons from the brainstem and cerebellum grow into the diencephalon early in development, but that they do not influence the cerebral cortex until relatively late. When the results of the present study are compared with those reported previously on the development of ascending spinal (Martin et al., '83) and corticofugal (Martin et al., '80; Cabana and Martin, '85b,c) projections, it appears that specific components of major somatosensory and motor circuits develop according to different timetables.

Animals↗

PubMed1985

Tamoxifen alone or in combination with estradiol-17 beta inhibits the growth and malignant transformation of hepatic hyperplastic nodules.

Hepatic hyperplastic nodules (HHNs) induced by the 'resistant hepatocyte method' of Solt et al. were studied as an experimental prototype of oral contraceptive-related tumors. Cytoplasmic estrogen receptors were present in all HHNs harvested and their concentration was always less than that in normal liver. No specific cytoplasmic progestin receptors could be measured in the above tumor or liver specimens. The long-term administration of estradiol-17 beta (4.8-24.0 micrograms/day) resulted in the death of all but one of 20 animals prior to termination at 10 months. Tamoxifen (0.25-2.5 mg biweekly), which did not lead to excess mortality, decreased HHN grade (proportion of liver slice occupied by HHN) and inhibited malignant transformation. Combination therapy with single-dose estradiol-17 beta (4.8 micrograms/day) and various doses of tamoxifen (0.25-2.5 mg biweekly) in most cases reduced mortality, HHN grade and malignant transformation. Cytoplasmic progestin receptors were absent and estrogen receptors were either undetectable or present in low concentrations in hepatic tumors harvested at the time of termination. Our results indicated that HHNs are hormone-dependent and that malignant transformation can be inhibited by tamoxifen alone or in combination with estradiol-17 beta.

Animals↗

PubMed1985

Tamoxifen alone or in combination with estradiol-17 beta inhibits the growth and malignant transformation of hepatic hyperplastic nodules.

Hepatic hyperplastic nodules (HHNs) induced by the 'resistant hepatocyte method' of Solt and Farber were studied as an experimental prototype of oral contraceptive-related tumors. Cytoplasmic estrogen receptors were present in all HHNs harvested and their concentration was always less than that in normal liver. No specific cytoplasmic progestin receptors could be measured in the above tumor or liver specimens. The long-term administration of estradiol-17 beta (4.8-24.0 micrograms/day) resulted in the death of all but one of 20 animals prior to termination at 10 months. Tamoxifen (0.25-2.5 mg biweekly) which did not lead to excess mortality, decreased HHN grade (proportion of liver slice occupied by HHN) and inhibited malignant transformation. Combination therapy with single-dose estradiol-17 beta (4.8 micrograms/day) and various doses of tamoxifen (0.25-2.5 mg biweekly) in most cases reduced mortality, HHN grade and malignant transformation. Cytoplasmic progestin receptors were absent and estrogen receptors were either undetectable or present in low concentration in hepatic tumors harvested at the time of termination. Our results indicate that HHNs are hormone-dependent and that malignant transformation can be inhibited by tamoxifen alone or in combination with estradiol-17 beta.

Animals↗

PubMed1983

Auditing antibiotic use in a teaching hospital: focus on cefoxitin.

The prevalence and nature of antibiotic misuse in a major teaching hospital was assessed by means of a quality-of-use audit. Cefoxitin was chosen for study. The use of cefoxitin increased rapidly during the study period, accounting for 17.7% ($15300) of the pharmacy's costs for cephalosporins during the first year of its availability but 47.7% ($60707) during the second year. Cefoxitin was inappropriately used for 43% of the patients receiving it during the first 2-month audit period and for 25% of those receiving it during the second audit period, 1 year later (p less than 0.01 by chi-square analysis).

Anti-Bacterial Agents↗

PubMed1983

Absorption of contrast medium during ERCP.

In order to assess systemic absorption, serum diatrizoate levels were measured in 25 patients requiring endoscopic retrograde cholangiopancreatography (ERCP). Urinary diatrizoate was measured in the urine of seven of these. In five additional patients, diatrizoate was instilled into the duodenum during endoscopy. Blood was drawn before and after completion of the procedure in all patients and at intervals from 2 to 24 hr in nine. Eighteen-hour urine collections were obtained from seven patients. Increases in serum diatrizoate concentration were 7.10 micrograms/ml +/- 3.01 (mean +/- SE) in the duodenal-instillation patients, 230.68 +/- 53.24 micrograms/ml in 9 patients in whom only the pancreatic duct was visualized, 7.83 +/- 1.05 micrograms/ml in three patients in whom only the bile duct was visualized, and 77.67 +/- 28.22 micrograms/ml in 13 patients in whom both ducts were visualized. The mean total urinary excretion of diatrizoate was 3.05 +/- 0.98 g of diatrizoate. Endoscopists performing ERCP should be aware of the likelihood of systemic absorption and the possibility of reactions to iodinated contrast materials in sensitive patients.

Absorption↗

PubMed1983