Enzymatic Capabilities of Bacteria Associated with Apple Maggot Flies: A Postulated Role in Attraction

Isolates of the bacterium Enterobacter agglomerans, obtained from the alimentary tract of the apple maggot fly, Rhagoletis pomonella, were found to be attractive or nonattractive in attraction tests. Polyacrylamide gel electrophoresis (PAGE) of whole bacterial cell and outer membrane preparations of bacteria revealed the presence of a protein confined to isolates attractive to the flies. This protein, with an approximate molecular weight of 106 kDa, was found to be extracellular in E. agglomerans, but absent in either whole cell or extracellular preparations of Klebsiella oxytoca or Klebsiella pneumoniae. Comparison of this protein with authentic uricase revealed that this protein has a molecular weight, a PAGE migration rate (R _f = 0.22), and behavior on a G-100 Sephadex column similar to pure uricase. This protein appears to be made by E. agglomerans in the absence of uric acid, i.e., constitutively, and its presence is positively linked to apple maggot fly response in attraction assays. The probable roles of this unique protein in purine degradation and the lives of apple maggot adults are discussed.     

Gastric emptying in control subjects and patients with duodenal ulcer before and after vagotomy

The emptying of a solid meal labelled with Indium 113mDTPA from the stomach was studied with a gamma camera in 26 normal subjects, 27 patients with duodenal ulcer, on 41 occasions after truncal vagotomy and pyloroplasty and 38 times after highly selective vagotomy. Applying the method of principal component analysis to the results, differences were detected between control and duodenal ulcer subjects and two probable subgroups of duodenal ulcer were observed. Half emptying times did not reveal these patterns. After vagotomy, delayed emptying was general at one week. At one month, patients after highly selective vagotomy had a more normal result than those with truncal vagotomy and pyloroplasty (TV), but by six months no significant difference in overall emptying rate was found, although changes in the pattern of gastric emptying persisted in some patients after TV.     

Recovery in rats after spinal cord injury

Previous studies from this laboratory have shown evidence of regeneration of long descending spinal motor tracts in rats after spinal cord transection and treatment to modify the animals' immune response. In this study, less extensive surgical lesions were combined with the most favorable drug treatment (75 mg per kilogram of cyclophosphamide in a single dose) in an effort to improve the prospects for regeneration. Less than complete spinal cord transections in the rat were frequently followed by clinical and electrophysiologic evidence of return of function. Such return of function appears to depend on a reorganization of the nervous system that results in the use of the few remaining fibers to transmit motor information rather than on regeneration. Immunosuppressive treatment had no effect on these results.     

Regulation of N-acetylglucosaminyltransferase V by protein kinases

When 7721 human hepatocarcinoma cells were treated with 100 nM phorbol-12-myristate-13-acetate (PMA), the activity of N-acetylglucosaminyltransferase V(GnT-V) in the cells varied in accordance with the activity of membranous protein kinase C (PKC), but not with that of cytosolic PKC. Quercetin, a non-specific inhibitor of Ser/Thr protein kinase, and D-sphingosine and staurosporine, two specific inhibitors of PKC, blocked the activation of membranous PKC and GnT-V by PMA. Among the three inhibitors, quercetin was least effective. The inhibitory rates of quercetin and staurosporine toward membranous PKC and GnTV were proportional to the concentrations of the two inhibitors. The activities of GnTV and membranous protein kinase A (PKA) were also induced in parallel by dibutyryl cAMP (db-cAMP) and this induction was blocked by a specific PKA inhibitor. When cell free preparations of 7721 cells and human kidney were treated with alkaline phosphatase (ALP) to remove the phosphate groups, the GnTV activities were decreased. These results suggest that GnTV may be activated by membranous PKC or PKA, indirectly or directly, via phosphorylation of Ser/Thr residues.     

Complement-derived anaphylatoxins in human donor corneas treated with excimer laser

BACKGROUND AND OBJECTIVE: An inflammatory response produced by excimer laser photorefractive keratectomy (PRK) may be associated with the subsequent corneal haze and regressions in refractive error observed after treatment. Complement-derived anaphylatoxins, potent mediators of inflammation, may have a role in postoperative healing. MATERIALS AND METHODS: Twenty right human donor corneas underwent a 6-D excimer laser PRK treatment. The corresponding left donor corneas served as the controls. After incubation in tissue culture media for 6 hours and elution in phosphate-buffered saline with EDTA for 24 hours, complement-derived anaphylatoxins C3a, C4a, and C5a were measured in corneal eluates by radioimmunoassay. RESULTS: Compared with control corneas, the excimer PRK corneas failed to demonstrate a significant increase in C3a, C4a, or C5a levels (P > .05). CONCLUSIONS: These results suggest that the excimer laser at this dose does not activate significant complement in the cornea.     

Dose-dependent gingival overgrowth induced by cyclosporin in rats

This study evaluated the effect of dosage on severity of cyclosporin-A (CSA) induced gingival overgrowth. Eighty (80) male Sprague-Dawley rats were randomly distributed into 4 groups. Rats in each group daily received CSA in mineral oil by gastric feeding at dosages of 0 (control), 3, 10, and 30 mg/kg, respectively, for 6 weeks. Stone models of the mandibular incisal region were obtained biweekly and were used for analysis of the gingival dimensions. Animals were sacrificed at the end of week 6 and tissue sections were processed for histopathologic evaluations. Animals were sacrificed at the end of week 6 and tissue sections were processed for histopathologic evaluation Gingival overgrowth including bucco-lingual and mesio-distal width and vertical height were significantly increased with increasing CSA dosage. Furthermore, the gingival dimensions displayed a positive linear relation to dosage and treatment duration. The histopathologic evaluation revealed a granulomatous tissue wedging the tooth-gingival interface in the 3 mg/kg group. This tissue had reached exuberant size in the 10 and 30 mg/kg groups. In summary, the analysis of gingival dimensions the histopathologic evaluation shows a dose-dependent effect on the severity of CSA-induced gingival overgrowth.     

Effects of parathyroid hormone (PTH)-related protein and PTH on osteoclasts and osteoclast precursors in vivo

Increased production of PTH-related protein (PTHrP) and PTH is frequently responsible for hypercalcemia and its associated morbidity. However, it is unclear whether these peptides produce identical effects on cells in the osteoclast lineage in vivo. To examine the effects of continuous in vivo exposure to these factors on both the osteoclast precursors and mature osteoclasts, we inoculated Chinese hamster ovarian cells expressing PTH-(1-84), PTHrP-(1-141), or nontransfected Chinese hamster ovarian cells into nude mice. The effects of these tumors on blood ionized calcium, plasma PTH and PTHrP concentrations, and osteoclast formation were then determined. PTH and PTHrP tumor-bearing mice became hypercalcemic (1.90 +/- 0.04 and 1.97 +/- 0.16 mmol/liter, respectively) compared with control mice (1.29 +/- 0.015 mmol/liter). After 4 days of hypercalcemia, mice were killed, and bone marrow cells were harvested to examine cells at three discrete stages of osteoclast development: multipotent osteoclast precursors, the granulocyte/macrophage colony-forming unit; more committed marrow mononuclear osteoclast precursors; and mature osteoclasts. Neither PTH nor PTHrP had an effect on granulocyte/macrophage colony-forming unit, but similarly increased the number of more committed mononuclear osteoclast progenitors as well as mature osteoclasts in the calvaria. No differences were detected between the effects of PTH and PTHrP on cells in the osteoclast lineage in vivo. Thus, PTH and PTHrP appear to affect only more differentiated cells in the osteoclast lineage, and the differences in osteoclastic bone resorption between primary hyperparathyroidism and humoral hypercalcemia of malignancy are probably due to mechanisms other than effects on osteoclast precursor cells in vivo.