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Biomedical subjects

J S Thompson

Publications and source records attributed to J S Thompson.

At least 199 records · Page 11Linked to original sources

Long-term effects of jejunoileal autotransplantation on myoelectrical activity in canine small intestine.

We studied the longitudinal effects of autotransplantation on the motor function of the jejunoileum. By performing the autotransplantation procedure in a manner similar to that employed for allotransplantation, we sought to examine the long-term effects of both extrinsic denervation and the operative procedure itself on small intestinal motor function. Although initially disrupted, interdigestive myoelectrical activity demonstrated progressive organization: 88% of migrating myoelectrical complexes in animals studied between 12 and 20 months following autotransplantation demonstrated each phase of the complex in normal sequence. Longitudinal studies of several parameters of myoelectrical activity provided further evidence of progressive organization and entrainment of motor functions within the denervated intestine. Several abnormal myoelectrical patterns were observed within the autotransplanted segment, however, and coordination of either slow wave or phase III activity with the proximal innervated intestine did not recover with time. The major component of the myoelectrical response to feeding was permanently impaired with a delayed onset and shortened duration of the fed response. We conclude that the extrinsically denervated intestine recovers the ability to generate and organize all phases of the MMC but demonstrates permanent impairment of the major motor response to food. However, anoxic and cooling damage to enteric nerves and muscle, incurred during the autotransplantation procedure, may explain the persistence of abnormal motor patterns and impaired myoelectrical conduction and could play an important role, additional to that of extrinsic denervation, in the long-term motor function of the allotransplanted intestine.

Animals↗

Neomucosal growth in serosa lined intestinal tunnels.

Functional intestinal mucosa, termed neomucosa, will grow in patched intestinal defects, but intestinal patching does not result in significantly increased intestinal surface area. Our aim was to determine if neomucosal growth would occur longitudinally in serosa-lined tunnels and to evaluate the optimal type of tunnel construction. Thirty rabbits had serosa-lined intestinal tunnels 1.5 cm in diameter and 5 cm in length interposed in the ileum. Group I (n = 5) had the tunnel formed from adjacent colon segments. Group II (n = 15) had a similar tunnel created with placement of an intraluminal latex stent. Group III (n = 5) had the tunnel formed by imbricating the cecum. Group IV (n = 5) had the tunnel formed by imbrication plus the intraluminal stent. Only Group II animals survived beyond 7 days (12 of 15 vs 0 of 5, P less than 0.05) and were sacrificed at 1 week (n = 3), 2 weeks (n = 3), 3 weeks (n = 2), 4 weeks (n = 2), and 6 weeks (n = 2). All other animals died from intestinal obstruction or peritonitis. Neomucosal growth occurred from proximal and distal margins. Forty percent of the tunnel was epithelialized at 6 weeks and the 5 cm tunnel contracted to 3 cm. In vitro glucose uptake was similar in neomucosa and normal mucosa but disaccharidase activity (sucrase and maltase) was significantly less in neomucosa at 6 weeks (28 +/- 35 vs 84 +/- 12 and 72 +/- 51 vs 471 +/- 84, P less than 0.05). Although not clinically obstructed, the survivors lost 15% body weight and proximal intestinal diameter increased.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

In-home pasteurization of raw goat's milk by microwave treatment.

Raw milk has been implicated as an important source of infection with the common bacterial agents associated with gastroenteritis. While established methods of home pasteurization can be cumbersome and tedious, we have demonstrated that a domestic microwave oven can be used effectively to reduce aerobic plate counts in raw goat's milk by up to 6 log cycles without impairing the organoleptic quality. Good keeping quality of the irradiated product was demonstrated by the 7-day holding standard plate count. Petrifilm SM and Petrifilm VRB were used successfully under field conditions. Further investigation must be undertaken to demonstrate that this method is equivalent to the legally accepted pasteurization procedures for the inactivation of such pathogenic bacteria as Salmonella spp., Listeria monocytogenes and the pasteurization standard Coxiella burnetti.

Animals↗

Glucagonoma syndrome is an underdiagnosed clinical entity.

Glucagonomas, considered among the rarest of the islet cell neoplasms, produce a well-defined clinical syndrome characterized by necrolytic migratory erythema, diabetes mellitus, glossitis, anemia, and weight loss. This report describes seven patients with glucagonoma treated at our institution. All seven had the characteristic dermatologic manifestations, present from 1 to 6 years prior to diagnosis. Five patients had extensive disease at the time of initial operation, three of whom underwent aggressive cytoreductive surgery, whereas the other two had biopsy only. The remaining two patients presented with a single nodule each, underwent distal pancreatectomy and splenectomy, and remain free of disease 2 and 6 years postoperatively. Earlier recognition of the distinctive physical findings peculiar to this syndrome should increase survival. Aggressive cytoreductive surgery results in prolonged remission.

Female↗

Repetitive strain injuries. How to deal with 'the epidemic of the 1990s'.

Occupational trends, especially the mushrooming of computer use in the United States, have brought with them a virtual epidemic of repetitive strain injury of the upper limb. What can you as a primary care physician do to stem the tide? In this article, the authors fill you in on treatment of the most common injuries as well as remark on the ramifications for business and industry.

Adult↗

Serum enzyme levels during intestinal ischemia.

Because the intestinal mucosa is most sensitive to ischemia, serum levels of mucosal enzymes, such as diamine oxidase, may be most likely to indicate intestinal ischemia. Our aim was to compare serum levels of mucosal (diamine oxidase, alkaline phosphatase) and seromuscular (creatinine phosphokinase, lactic dehydrogenase, serum glutamic oxaloacetic transminase) enzymes during intestinal ischemia of varying extent and duration in dogs. Group 1 (n = 6) underwent sham laparotomy. Group 2 (n = 8) had 50% of the small intestine devascularized. Group 3 (n = 8) had the superior mesenteric artery occluded for 2 hours and released. Group 4 (n = 8) had the superior mesenteric artery ligated. Serum samples were obtained before and 2, 4, 8, and 24 hours after operation, and histologic specimens were examined at 4 hours. Creatinine phosphokinase levels became elevated within 4 hours of ischemic injury in group 2 (223 +/- 197 vs. 68 +/- 26, p less than 0.05) and group 4 (212 +/- 136 vs. 76 +/- 29, p less than 0.05). Significant elevation of serum enzymes levels, except diamine oxidase, occurred in groups 2, 3, and 4 at 24 hours, including those with normal histology after temporary superior mesenteric artery occlusion. Thus seromuscular enzymes, particularly creatinine phosphokinase, were more likely to be elevated during intestinal ischemia. Enzyme levels were not influenced by the extent and reversibility of the ischemic injury.

Alkaline Phosphatase↗

Basement membrane components stimulate epithelialization of intestinal defects in vivo.

Subepithelial tissues have an important role in the structure and function of the intestinal epithelium. Basement membrane components (BMC) stimulate epithelial cell migration and differentiation in vitro. The aim of this study was to determine the effect of BMC and/or interstitial tissue collagen (type I) on the in vivo intestinal regenerative response to intestinal patching. Twenty rabbits had two 2 x 5 cm ileal defects patched with the serosal surface of adjacent caecum. Group 1 (n = 5) were controls; group 2 (n = 5), group 3 (n = 5) and group 4 (n = 5) had collagen, collagen plus BMC, and BMC respectively applied to the distal patched defect. Animals were killed at 7 d and evaluated grossly for epithelialization and contraction of the defects. Epithelial coverage was greatest in the distal patch of group 4 animals (62 +/- 9%) and was significantly greater than the group 4 proximal patch and control values (43 +/- 7 and 40 +/- 14%, P less than 0.05). Contraction was similar in all groups (38 +/- 5 to 45 +/- 5%). Crypt cell production rate, villus height, and disaccharidase activities were similar in all groups. BMC stimulated epithelialization via a local mechanism since only the distal patch was affected. Type I collagen did not stimulate epithelialization and inhibited the effect of BMC. Since crypt cell production rate was similar in all groups, the enhanced epithelialization seen with BMC is primarily due to increased cell migration.

Animals↗

Sequencing the gene for an imipenem-cefoxitin-hydrolyzing enzyme (CfiA) from Bacteroides fragilis TAL2480 reveals strong similarity between CfiA and Bacillus cereus beta-lactamase II.

Using a newly constructed Bacteroides fragilis-Escherichia coli cloning shuttle vector, pJST61, we have cloned the cefoxitin (FOX)-imipenem (IMP) resistance determinant from B. fragilis TAL2480. FOX-IMP resistance in this strain results from the production of a periplasmic, Zn2(+)-containing beta-lactamase which hydrolyzes carbapenems and cephamycins and whose activity is resistant to clavulanic acid but sensitive to Zn2(+)-binding reagents, including EDTA. The pJST61 vector permits efficient library construction in E. coli and allows for the transfer of the library to B. fragilis recipients for the screening or selection of specific phenotypes. The library clone containing the FOX-IMP resistance gene was detected after transfer to B. fragilis TM4000 (Fox-Imps) selecting for Foxr. One of the isolates carrying plasmid pJST241 is resistant to FOX and IMP and synthesizes a periplasmic protein with substrate and inhibitor properties identical to those of strain TAL2480. On the basis of deletion analysis, Tn1000 insertion mutations, and DNA sequencing, we have defined the 747-base cfiA (FOX-IMP resistance) gene within the 3.6-kilobase cloned insert in pJST241. The cfiA gene contains an open reading frame that could code for a precursor protein of 249 amino acids and with a molecular mass of 27,260 daltons. A potential signal sequence has been identified at the N terminus of this protein; cleavage within this sequence would result in a protein of 231 amino acids with a molecular mass of 25,249 daltons. The CfiA protein shows remarkable similarities to the exported, Zn2(+)-requiring, type II beta-lactamase Blm proteins from Bacillus cereus 569/H and 5/B/6. Although overall amino acid identity is only 32%, the Zn ligand-binding His and Cys residues are precisely conserved and the amino acids in the vicinity of these sites show strong similarities (greater than 80%) when the CfiA and Blm proteins are compared.

Amino Acid Sequence↗

Cloning and expression of the Bacteroides fragilis TAL2480 neuraminidase gene, nanH, in Escherichia coli.

We have cloned the Bacteroides fragilis TAL2480 neuraminidase (NANase) structural gene, nanH, in Escherichia coli. This was accomplished by using the cloning shuttle vector pJST61 and a partial Sau3A library of TAL2480 chromosomal inserts created in E. coli. The library was mobilized into the NANase-deficient B. fragilis TM4000 derivative TC2. NANase-producing colonies were enriched by taking advantage of the inability of TC2, but not the wild-type of NANase+ revertant, to grow in vitro in fluid aspirated from the rat granuloma pouch. Plasmids pJST61-TCN1 and pJST61-TCN3, containing inserts of 9.1 and 4.5 kilobases (kb), respectively, were found in the TC2 derivatives that grew in the rat pouch medium. In B. fragilis, NANase production from the two plasmids was inducible by free N-acetylneuraminic acid or sialic acid-containing substrates, just as in the parental TAL2480 strain. However, when these plasmids were transferred back to E. coli, NANase activity was barely detectable. A 3.5-kb portion of the insert in pJST61-TCN3 was subcloned in pJST61 to give plasmid pJST61-SC3C; NANase was produced from this plasmid both in E. coli and in B. fragilis. In E. coli, NANase expression was under the control of the vector promoter lambda pR and was therefore completely abolished by the presence of a lambda prophage. In B. fragilis, NANase production was inducible by free N-acetylneuraminic acid or sialic acid-containing substrates. By using deletion analysis and Tn1000 mutagenesis, the NANase structural gene and control region that functions in B. fragilis were localized to a 1.5- to 2.0-kb region of the insert. A partial nucleotide sequence of the NANase-deficient Tn1000 insertion mutants allowed us to identify the nanH gene and deduce the amino acid sequence of a portion of the NANase protein. We identified five regions showing great similarity to the Asp boxes, -Ser-X-Asp-X-Gly-X-Thr-Trp-, of other bacterial and viral NANase proteins.

Amino Acid Sequence↗

Rapid biochemical test to identify verocytotoxin-positive strains of Escherichia coli serotype O157.

Fluorogenic procedures were used with the substrate 4-methylumbelliferyl-beta-D-glucuronide (MUG) to identify Escherichia coli. Most strains produced beta-glucuronidase and, thus, were MUG positive. A 20-min procedure was developed to detect glucuronidase activity in 1,295 bacterial cultures, representing 23 genera, of strains that were isolated from clinical specimens. Very few organisms other than E. coli were MUG positive. Of 682 E. coli strains that were isolated, 630 (92.4%) were MUG positive. When an additional 188 E. coli serotype O157 isolates were examined, 155 E. coli O157:H7, 10 E. coli O157:H-, and 1 E. coli O157:H (rough) isolate were MUG negative. All 166 cultures were verocytotoxin positive. Of the remaining 22 E. coli O157 isolates, 2 isolates were O157:H-, 1 isolate was O157:H (rough), and 19 isolates were other H types (H6, H16, H19, H25, H42, and H45); these 22 isolates were MUG positive. All 22 cultures were verocytotoxin negative. The rapid MUG procedure can be used to predict verocytotoxin-positive isolates of E. coli O157; that is, there is a very good likelihood that MUG-negative E. coli O157 isolates are verocytotoxin positive.

Bacterial Toxins↗

Use of tri-gas incubator for routine culture of Campylobacter species from fecal specimens.

We evaluated a tri-gas incubator for Campylobacter isolation to be used instead of an anaerobic jar. Fecal specimens were cultured in duplicate onto charcoal selective medium and incubated at 43 degrees C for 48 h in two different environments: a tri-gas incubator (Forma Scientific) adjusted to provide an atmosphere of 10% CO2, 10% O2, and the balance N2; and evacuated anaerobic jars with a replacement gas mixture of 10% CO2, 5% O2, and 85% N2. A total of 106 Campylobacter jejuni and 8 Campylobacter coli isolates were obtained from 2,348 stool specimens. Of the positive specimens, 113 isolates came from the incubator and 111 isolates came from the anaerobic jars. An additional 32 previously positive specimens were replated onto charcoal selective medium and retested by both methods. We recovered 27 C. jejuni isolates, 26 isolates by each method. The isolates from the incubator typically produced discrete colonies, while the isolates from the anaerobic jar showed some degree of swarming in colony formation. The tri-gas incubator provided a cost-effective method for culturing Campylobacter spp.

Bacteriological Techniques↗

Comparison of ulcer surgery at a Veterans Administration and University Hospital.

We compared the patient populations and outcome of surgery for peptic ulcer disease in 81 patients at a Veterans Administration Hospital (OVAH) and 97 patients at an affiliated University Hospital (UNH). The surgeons and choice of operation were comparable at both facilities. Patients were similar with respect to severity of ulcer disease, percentage of elderly patients and distribution of comorbid conditions. There were significantly more female patients (45% vs 2%, P less than 0.05), patients less than 40 years old (22% vs 6%, P less than 0.05), and patients with gastric ulcers (27% vs 12%, P less than 0.05) at UNH. Alcoholism was more prevalent at OVAH (57% vs 22%, P less than 0.05). Postoperative morbidity and mortality rates were 27 per cent and 14 per cent at OVAH and 25 per cent and 16 per cent at UNH. These data suggest that quality of care for surgical therapy of ulcer disease at a VA Hospital is comparable to its tertiary care affiliate with a similar patient population.

Adult↗

Hyperammonemic encephalopathy in urinary diversion with urea-splitting urinary tract infection.

We present two cases of hyperammonemic encephalopathy secondary to urea-splitting urinary tract infection with urinary diversion. One patient had a ureterosigmoidostomy, the other an ileal loop diversion. Neither patient had significant underlying liver disease, but both had considerable muscle atrophy that may have predisposed them to develop hyperammonemia. Medical therapy did not provide long-term control of symptoms. In both cases, hyperammonemic encephalopathy resolved after revision of their urinary diversions. The probable mechanism of the metabolic derangements produced by urea-splitting urinary tract infections is reviewed. We suggest that patients with urinary diversion who develop hyperammonemic encephalopathy secondary to a urea-splitting urinary tract infection be treated with surgical revision of the urinary system to improve drainage and decrease bowel contact time.

Acidosis↗

Characterization of the inhibitor complexes of cobalt carboxypeptidase A by electron paramagnetic resonance spectroscopy.

The metal coordination sphere of cobalt-substituted carboxypeptidase A and its complexes with inhibitors has been characterized by X-band electron paramagnetic resonance (EPR) spectroscopy. The temperature dependence of the EPR spectrum of cobalt carboxypeptidase and the g anisotropy are consistent with a distorted tetrahedral geometry for the cobalt ion. Complexes with L-phenylalanine, a competitive inhibitor of peptide hydrolysis, as well as other hydrophobic L-amino acids all exhibit very similar EPR spectra described by three g values that differ only slightly from that of the cobalt enzyme alone. In contrast, the EPR spectra observed for the cobalt enzyme complexes with 2-(mercaptoacetyl)-D-Phe, L-benzylsuccinate, and L-beta-phenyllactate all indicate an approximately axial symmetry of the cobalt atom in a moderately distorted tetrahedral metal environment. Phenylacetate, beta-phenylpropionate, and indole-3-acetate, which exhibit mixed modes of inhibition, yield EPR spectra indicative of multiple binding modes. The EPR spectrum of the putative 2:1 inhibitor to enzyme complex is more perturbed than that of the 1:1 complex. For beta-phenylpropionate, partially resolved hyperfine coupling (122 x 10(-4) cm-1) is observed on the g = 5.99 resonance, possibly indicating a stronger metal interaction for this binding mode. The structural basis for the observed EPR spectral perturbations is discussed with reference to the existing crystallographic kinetic and electronic absorption, nuclear magnetic resonance, and magnetic circular dichroic data.

Carboxypeptidases↗

Effect of eflorithine on intestinal regeneration.

Patching intestinal defects with adjacent serosal surfaces results in the growth of new intestinal mucosa. Since polyamine biosynthesis is associated with cellular growth and differentiation, it may be important in this regenerative process. Our aim was to determine the effect of eflorithine (difluoromethylornithine), a specific inhibitor of polyamine synthesis, on intestinal regeneration. Forty-eight New Zealand white rabbits had 2 x 5-cm ileal defects patched with adjacent cecal serosal surface. One half of the animals took 2% eflorithine in drinking water postoperatively. Six animals in each group were killed 7, 14, 21, and 28 days after patching. There was no significant difference in neomucosal growth at any time. Villous height, disaccharidase activity, and crypt cell production were significantly lower in the eflorithine-treated animals. Eflorithine-treated animals had significantly lower ornithine decarboxylase activity and polyamine levels. Despite the inhibitory effect of eflorithine on polyamine synthesis and proliferative activity, epithelialization and contraction of the patched defect were not affected. These findings suggest that polyamine synthesis is important in proliferation and differentiation of cells in the neomucosa but does not influence cell migration in intestinal regeneration.

Animals↗