Biomedical subjects
Publications and source records attributed to C D Smith.
A 39-year-old woman presented with an acute left hemisphere stroke and was found to have severe hyperthyroidism. She developed an acute right brachial artery occlusion while under continuous cardiac monitoring, and at no time exhibited atrial fibrillation. This case, and a review of the literature, suggests that thromboembolism occurs in thyrotoxicosis without atrial fibrillation.
A 34-year-old man had progressive, poorly controlled low back pain for 3 years before testing confirmed a diagnosis of ochronosis. This is the first case description of the magnetic resonance imaging (MRI) appearance of spinal ochronosis, which should be considered when advanced degenerative change is seen on spinal MRI in young individuals.
A woman with previously treated disseminated histoplasmosis was admitted with progressive paraparesis. Magnetic resonance imaging showed multifocal cerebritis and spinal histoplasmoma. She recovered following a second treatment with amphotericin B. This is the fourth reported patient with intramedullary spinal histoplasmoma, and the first demonstrating successful treatment of the lesion with medical therapy alone.
Two long terminal repeat (LTR) enhancer-binding proteins which may regulate high rates of avian leukosis virus (ALV) LTR-enhanced c-myc transcription during bursal lymphomagenesis have been identified (A. Ruddell, M. Linial, and M. Groudine, Mol. Cell. Biol. 9:5660-5668, 1989). The genes encoding the a1/EBP and a3/EBP binding factors were cloned by expression screening of a lambda gt11 cDNA library from chicken bursal lymphoma cells. The a1/EBP cDNA encodes a novel leucine zipper transcription factor (W. Bowers and A. Ruddell, J. Virol. 66:6578-6586, 1992). The partial a3/EBP cDNA clone encodes amino acids 84 to 313 of vitellogenin gene-binding protein (VBP), a leucine zipper factor that binds the avian vitellogenin II gene promoter (S. Iyer, D. Davis, and J. Burch, Mol. Cell. Biol. 11:4863-4875, 1991). Multiple VBP mRNAs are expressed in B cells in a pattern identical to that previously observed for VBP in other cell types. The LTR-binding activities of VBP, a1/EBP, and B-cell nuclear extract protein were compared and mapped by gel shift, DNase I footprinting, and methylation interference assays. The purified VBP and a1/EBP bacterial fusion proteins bind overlapping but distinct subsets of CCAAT/enhancer elements in the closely related ALV and Rous sarcoma virus (RSV) LTR enhancers. Protein binding to these CCAAT/enhancer elements accounts for most of the labile LTR enhancer-binding activity observed in B-cell nuclear extracts. VBP and a1/EBP could mediate the high rates of ALV and RSV LTR-enhanced transcription in bursal lymphoma cells and many other cell types.
New drugs for causal prophylaxis of malaria are needed. A proguanil/sulfamethoxazole combination was investigated using a rhesus monkey model (Macaca mulatta infected with Plasmodium cynomolgi) to determine whether causal prophylaxis could be achieved. When a five-day regimen of proguanil (40 mg/kg/day) combined with sulfamethoxazole (100 mg/kg/day) was used, infection of all animals (6 of 6) was observed, with an extended prepatent period (median 40 days). Two control animals became infected on days 9 and 23 following sporozoite inoculation. Plasma concentrations indicated that proguanil and sulfamethoxazole were adequately absorbed and metabolized to cycloguanil and N4-acetylsulfamethoxazole, respectively. Analysis of liver biopsy specimens demonstrated that the drugs were present two days following sporozoite inoculation but were not detectable one week later. Proguanil plus sulfamethoxazole does not eliminate exoerythrocytic-stage parasites in the rhesus monkey--P. cynomolgi model.
The effects of a novel porphyrin, tolyporphin, on P-glycoprotein-mediated multiple drug resistance in human ovarian and breast cell lines were characterized. Compared with parental SKOV3 and MCF-7 cells, the P-glycoprotein-overexpressing sublines SKVLB1 and MCF-7/ADR were 5- and 1.3-fold less sensitive to the cytotoxic effects of tolyporphin. Subtoxic doses of tolyporphin increased the sensitivity of the SKVLB1 and MCF-7/ADR cells to P-glycoprotein-transported drugs, but did not increase the antiproliferative effects of nontransported drugs. Tolyporphin also enhanced the accumulation of [3H]-vinblastine in SKVLB1 and MCF-7/ADR cells at doses approximately 10-fold lower than those required for similar responses to verapamil. In contrast, tolyporphin did not affect drug accumulation in SKOV3 or MCF-7 cells. Tolyporphin reduced [3H]-vinblastine efflux from SKVLB1 cells, reduced [3H]-vinblastine binding to membranes from SKVLB1 cells, and blocked the ability of [3H]-azidopine to photoaffinity-label P-glycoprotein in these membranes. These results indicate that tolyporphin binds to P-glycoprotein and inhibits the transport of cytotoxic natural product drugs. This novel natural product, and related compounds, may be useful for the reversal of multiple drug resistance and for further definition of the drug binding site(s) of P-glycoprotein.
Following rodent surveys in a rice-growing area of central Thailand where we found Bandicota savilei, B. indica, and Rattus rattus infected with Rickettsia tsutsugamushi, we performed a study of pathogenesis of R. tsutsugamushi in laboratory-reared B. savilei. Eight animals were injected with saline and 19 animals were injected with 4.0 x 10(6) mouse 50% lethal dose units of a strain of R. tsutsugamushi isolated from a human in central Thailand. Animals were evaluated at intervals for IgG and IgM antibodies to R. tsutsugamushi by an indirect immunoperoxidase assay, the presence of the pathogen in liver and spleen by murine inoculation, and the pathology of representative tissues by gross and microscopic examination. The infected animals began to show internal evidence of mild illness 7-14 days after inoculation, and exhibited no changes in behavior. Total white blood cell counts decreased on day seven (including lymphocytes and polymorphonuclear leukocytes), followed by an almost equal increase on day 14. Gross pathology noted at necropsy was limited to slight liver and spleen enlargement accompanied by low numbers of abscesses and fibrinous tags present in the abdominal cavity. In addition to the gross morphologic changes, histopathologic lesions noted were all mild, consisting of vasculitis of the lung, activation of the mononuclear phagocyte system, abdominal mesothelial cell hyperplasia, and peritonitis. Rickettsiae were isolated from liver and spleen on days 0, 7, and 14, but not thereafter. Specific antibody response was first observed on day 14, peaked on day 21, and it decreased to levels observed in uninfected animals between days 120 and 180.(ABSTRACT TRUNCATED AT 250 WORDS)
BACKGROUND: Our aim was to assess changes in transport of water and electrolytes under basal, proabsorptive, and prosecretory conditions after an in situ neural isolation of the jejunoileum. METHODS: Eight dogs underwent intraoperative perfusion of 50 cm of jejunum with a balanced electrolyte solution during sham operation and after neural isolation of the jejunoileum. Jejunal perfusion studies were later conducted in seven conscious dogs with a triple-lumen technique before and 2 and 8 weeks after neural isolation of the jejunoileum during intravenous infusion of 150 mmol/L sodium chloride (basal conditions), vasoactive intestinal polypeptide (VIP conditions), 500 pmol/kg per hour (prosecretory conditions), and the alpha 2-adrenergic agonist alpha-methylnorepinephrine (MNE), 900 nmol/kg per hour (proabsorptive conditions). RESULTS: Neural isolation decreased intraoperative net absorption of water (4.4 +/- 0.9 vs 2.2 +/- 0.5 microliters/cm/min; p < 0.05) and electrolytes (sodium, chloride, bicarbonate, potassium). In conscious dogs during basal conditions, net absorptive flux was decreased (p < 0.05) at 2 but not at 8 weeks. VIP produced similar absolute decreases in net absorptive flux at all three time points. MNE increased net absorption before and at 8 weeks, but not at 2 weeks after autotransplantation. CONCLUSIONS: In situ neural isolation of the jejunoileum decreased net basal jejunal absorption of water and electrolytes immediately and for at least 2 weeks. Proabsorptive responses to MNE but not prosecretory responses to VIP were altered at 2 weeks. Jejunal adaptation allowed absorptive function to return to near normal by 8 weeks.
Avian leukosis virus (ALV) induces bursal lymphoma in chickens after rare integration of proviral long terminal repeat (LTR) sequences next to the c-myc proto-oncogene. Labile LTR-binding proteins are essential for c-myc hyperexpression, since LTR-enhanced transcription, as well as the activity of the LTR-binding proteins, is specifically decreased following protein synthesis inhibition. This lability is restricted to hematopoetic cells of ALV-susceptible chicken strains, suggesting that the labile proteins play an important role in susceptibility to lymphomagenesis. Five proteins that interact with the ALV LTR enhancer were identified from bursal lymphoma cells: two proteins are stable (b and c), while three proteins (a1, a3, and b*) are labile in pre-B cell types. Two cDNAs (a1/EBP and a3/EBP) encoding leucine zipper proteins that bind to the ALV LTR enhancer were cloned using a lambda gt11 cDNA expression library made from bursal lymphoma cells. We are presently studying the roles of a1/EBP and a3/EBP in labile regulation of LTR-enhanced c-myc transcription and susceptibility to bursal lymphomagenesis.
We present here the three-dimensional structure of neuraminidase (E.C. 3.2.1.18) from influenza virus A/Tern/Australia/G70c/75 (N9), determined by the method of multiple isomorphous replacement, and the structure of the neuraminidase complexed with an inhibitor, 2-deoxy-2,3-dehydro-N-acetyl neuraminic acid (DANA). Native and inhibitor complex crystals are isomorphous and belong to space group I432 with unit cell dimensions of 183.78 A. The native enzyme structure and the inhibitor complex structure have been refined at 2.5 A and 2.8 A resolution, respectively, with crystallographic R-factor values of 0.193 for the native enzyme, and 0.179 for the inhibitor complex. The current enzyme model includes 387 amino acid residues which comprise the asymmetric unit. The root-mean-square deviation from ideal values is 0.013 A for bond lengths and 1.6 degree for bond angles. The neuraminidase (NA), as proteolytically cleaved from the virus, retains full enzymatic and antigenic activity, and is a box-shaped tetramer with edge lengths of 90 A and a maximal depth of 60 A. The NA tetramers are composed of crystallographically equivalent monomers related by circular 4-fold symmetry. Each monomer folds into six antiparallel beta-sheets of four strands. The secondary structure composition is 50% beta-sheet. The remaining 50% of the residues form 24 strand-connecting loops or turns. One of the loops contains a small alpha-helix. The structure of the complex of NA with DANA, a transition state analog, has enabled us to identify and characterize the site of enzyme catalysis. The center of mass of bound inhibitor is 32 A from the 4-fold axis of the tetramer, lodged at the end of a shallow crater of diameter 16 A with a depth of 8 to 10 A. There are 12 amino acid residues that directly bind DANA, with a further six conserved amino acids lining the active site pocket. The neuraminidase inhibitor complex provides a three-dimensional model which will be used to further the understanding of enzymatic hydrolysis and aid the design of specific, antineuraminidase antiviral compounds.
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Cells demonstrating the multidrug resistance phenotype because of overexpression of P-glycoprotein, a drug efflux pump, are resistant to the cytotoxic effects of most natural product drugs. To determine if P-glycoprotein confers resistance to the syctophycins, a family of natural cytotoxic macrolides recently isolated from cyanobacteria of the family Syctone-mataceae, we have characterized the effects of these compounds on drug-sensitive (SKOV3) and drug-resistant (SKVLB1) human ovarian carcinoma cells. While SKVLB1 cells demonstrated > 150- and 10,000-fold decreases in sensitivity to Adriamycin and vinblastine, respectively, they were equally sensitive as SKOV3 cells to the antiproliferative effects of tolytoxin and certain related scytophycins. The SKVKB1 cells were 4- to 11-fold resistant to other scytophycins and were 14-fold resistant to cytochalasin B. Microfilaments in SKOV3 and SKVLB1 cells were depolymerized by similar concentrations of tolytoxin, while cytochalasin B was less potent toward SKVLB1 cells than SKOV3 cells. Both tolytoxin and cytochalasin B enhanced the cytotoxicity of vinblastine toward SKVLB1 cells; however, neither compound affected the sensitivity to Adriamycin or cisplatin. Verapamil markedly increased the accumulation of [3H]vinblastine by SKLVB1 cells, while cytochalasin B caused only modest increase, and tolytoxin had no effect on [3H]vinblastine accumulation. These results suggest that some of the scytophycins, including tolytoxin, are not subject to P-glycoprotein-mediated efflux from cells exhibiting multidrug resistance due to overexpression of this transport protein. These compounds may therefore be useful for killing drug-resistant tumor cells.
Tolytoxin, a cytostatic, antifungal macrolide produced by blue-green algae of the genus Scytonema, is a potent, reversible inhibitor of cytokinesis in cultured mammalian cells. Treatment of KB cells with 2-16 nM tolytoxin results in profound morphological changes, beginning with the formation of zeiotic processes and culminating in nuclear protrusion. In L1210 cells, cytokinesis is inhibited by as little as 2 nM tolytoxin, while karyokinesis proceeds normally, resulting in polynucleation. Tolytoxin specifically disrupts microfilament organization in A10 cells, while having no apparent effect on microtubules or intermediate filaments. Tolytoxin inhibited actin polymerization in vitro and also caused the depolymerization or fragmentation of F-actin in vitro. Tolytoxin exhibits effects that closely resemble those of cytochalasin B but is effective at concentrations 1/50-1/1,000 that of cytochalasin B.
Although alpha 2-adrenergic agonists stimulate absorption in the mammalian small and large intestine in vitro, the possibility of central neural effects have confounded interpretation of in vivo studies. Our aim was to assess the effects of intravenous administration of alpha-methylnorepinephrine (MNE), an alpha 2-adrenergic agonist that does not cross the blood-brain barrier, on net jejunal absorption of water and electrolytes in the neurally intact, conscious dog. Absorption from a 30-cm proximal jejunal segment was studied using a triple-lumen perfusion technique in seven dogs. A warmed, isosmolar, balanced electrolyte solution containing [14C]polyethylene glycol was infused at 5 ml/min. Net jejunal fluxes of water and electrolytes were determined before, during, and after a 1.5-hr infusion of MNE (900 nmol/kg/hr). MNE increased net jejunal water absorption (from 12.9 +/- 1.8 to 22.5 +/- 1.5 microliters/cm/min, P < 0.05). Peripheral alpha 2-adrenergic receptors mediate a net proabsorptive response in the neurally intact canine jejunum in vivo independent of direct central neural effects.
To investigate metabolic disturbances in an animal model of human malaria, four rhesus monkeys (Macaca mulatta) were infected with Plasmodium coatneyi, a parasite which induces cytoadherence of infected erythrocytes. When moribund or the parasitaemia had plateaued, the monkeys were sacrificed (3 animals) or treated with chloroquine (1 animal). Blood and cerebrospinal fluid (CSF) were sampled at intervals between inoculation and sacrifice or treatment. Arterial and CSF glucose and lactate rose during infection, indicating evolving insulin resistance. The arteriovenous difference in glucose concentration also increased, consistent with increased glucose consumption by parasitised tissues. Arterial plasma lactate rose but a positive arteriovenous concentration difference suggested tissue lactate uptake. The animal with the highest plasma lactate at sacrifice remained hyperglycaemic but also had the highest CSF lactate, the greatest cerebral sequestration and neurological depression, and biochemical and histological evidence of severe hepatic pathology. Serum cholesterol and corrected serum calcium fell consistently during infection; serum phosphate was also reduced in animals without renal impairment. These preliminary results indicate that lactic acidosis is a late complication of severe malaria and, by implication from this and other studies, hypoglycaemia occurs even later; other metabolic changes during P. coatneyi infection in rhesus monkeys also parallel those of severe falciparum malaria in humans. The model could be used in further studies of malaria-associated metabolic dysfunction and its management.
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Serology to detect antibodies to Helicobacter pylori is not frequently used as a diagnostic tool in developing countries. When compared to a commercial ELISA, an ELISA constructed and validated in Thailand had a higher sensitivity (98% vs. 85%), specificity (76% vs. 66%), and negative predictive value (97% vs. 76%) for the detection of H. pylori infection among 104 patients with dyspepsia evaluated by endoscopy. The positive predictive value was 88% for both tests. Serum antibody levels fell significantly 5-8 months after eradication of infection in 8 Thai patients (P = .009). By 8 years of age, > 50% of Thai persons living in urban and rural locations were seropositive. The low negative predictive value of the commercial ELISA limits the usefulness of this assay as a diagnostic tool in Thailand and suggests a need to reevaluate H. pylori serologic tests when used in populations living in developing countries.
OBJECTIVE: To determine the local prevalence and optimal therapy for children with Barrett's esophagus (BE), the authors studied children with esophageal strictures or gastroesophageal reflux (GER), or both, to diagnose BE and to follow after therapy. SUMMARY BACKGROUND DATA: Barrett's esophagus is seldom reported in children and therapeutic recommendations are unclear. Barrett's esophagus usually develops during the mucosal reparative process after acid-reflux injury to the esophageal mucosa. Risk factors for BE include conditions that are associated with GER such as mental retardation, esophageal stricture, esophageal atresia, and reversed gastric tube esophagoplasty. Barrett's syndrome increases the risk of esophageal adenocarcinoma by 30 to 40 times. METHODS: All children with the risk factors had repeated esophagoscopy and multiple mucosal biopsies before and after therapy. RESULTS: Eleven children have been documented with BE. The initial diagnoses were: GER, 5; esophageal atresia, 4; nasogastric intubation, 1; lye ingestion, 1. A gastric tube esophagoplasty had been performed in three patients with BE in the esophagus proximal to the anastomosis. Three children with mid-esophageal strictures and long segments of BE had total resection with colic interposition. An additional two patients with tight stricture were treated with colic-patch esophagoplasty without resection. The final three patients were treated with fundoplication alone. CONCLUSIONS: Barrett's esophagus can be caused by acid from gastric tubes but responds to H2 blockers and diet. Resection eliminates BE; esophagoplasty only controls the stricture and must be accompanied by fundoplication. Barrett's esophagus persists in patients with fundoplication alone if reflux control is incomplete. The authors conclude that acid reflux must be controlled to treat BE successfully or the involved segment must be resected. Esophagogastrostomy apparently predisposes to BE.