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L C Kelley

Publications and source records attributed to L C Kelley.

At least 19 recordsLinked to original sources
PubMed2001

The cyclin-dependent kinases cdk2 and cdk5 act by a random, anticooperative kinetic mechanism.

cdk2.cyclin E and cdk5.p25 are two members of the cyclin-dependent kinase family that are potential therapeutic targets for oncology and Alzheimer's disease, respectively. In this study we have investigated the mechanism for these enzymes. Kinases catalyze the transfer of phosphate from ATP to a protein acceptor, thus utilizing two substrates, ATP and the target protein. For a two-substrate reaction, possible kinetic mechanisms include: ping-pong, sequential random, or sequential ordered. To determine the kinetic mechanism of cdk2.GST-cyclin E and cdk5.GST-p25, kinase activity was measured in experiments in which concentrations of peptide and ATP substrates were varied in the presence of dead-end inhibitors. A peptide identical to the peptide substrate, but with a substitution of valine for the phosphoacceptor threonine, competed with substrate with a K(i) value of 0.6 mm. An aminopyrimidine, PNU 112455A, was identified in a screen for inhibitors of cdk2. Nonlinear least squares and Lineweaver-Burk analyses demonstrated that the inhibitor PNU 112455A was competitive with ATP with a K(i) value of 2 microm. In addition, a co-crystal of PNU 112455A with cdk2 showed that the inhibitor binds in the ATP binding pocket of the enzyme. Analysis of the inhibitor data demonstrated that both kinases use a sequential random mechanism, in which either ATP or peptide may bind first to the enzyme active site. For both kinases, the binding of the second substrate was shown to be anticooperative, in that the binding of the first substrate decreases the affinity of the second substrate. For cdk2.GST-cyclin E the kinetic parameters were determined to be K(m, ATP) = 3.6 +/- 1.0 microm, K(m, peptide) = 4.6 +/- 1.4 microm, and the anticooperativity factor, alpha = 130 +/- 44. For cdk5.GST-p25, the K(m, ATP) = 3.2 +/- 0.7 microm, K(m, peptide) = 1.6 +/- 0.3 microm, and alpha = 7.2 +/- 1.8.

Adenosine Triphosphate↗
PubMed2001

Immunohistochemistry of ultimobranchial thyroid carcinomas in seven slaughtered cows and one bull.

Eight thyroid gland epithelial tumors were found in 7 cows and 1 bull in a retrospective study of thyroid gland lesions in slaughtered cattle. All tumors were classified as ultimobranchial thyroid carcinomas based on morphology and immunohistochemistry. All tumors consisted of solid sheets and nests of polygonal to oval epithelial cells, with more sparsely dispersed colloid-filled follicles. Connective tissue separating nests of epithelial cells varied from delicate fibrovascular stroma to dense collagenous stroma. Fusiform epithelial cells with rare neural fibers and ganglion cells were present in 1 tumor. Cells within solid areas of these tumors were immunoreactive for calcitonin, calcitonin gene-related peptide, neuron-specific enolase, and synaptophysin. Colloid and follicle cells were immunoreactive for thyroglobulin. Few follicle cells also were reactive for calcitonin gene-related peptide. Neoplastic cells invaded the fibrous capsules in all 8 cattle. These tumors represented proliferation of a mixed population of undifferentiated cells, C cells, and thyroid follicular epithelial cells, presumably derived from the thyroid ultimobranchial bodies. These ultimobranchial carcinomas in slaughtered cattle are comparable to ultimobranchial tumors described in dairy bulls and the intermediate type of thyroid gland carcinomas (mixed thyroid medullary carcinomas) described in human beings.

Animals↗
PubMed2000

A rapid DNA isolation procedure for the identification of Campylobacter jejuni by the polymerase chain reaction.

We have developed an efficient process for rapidly isolating campylobacter DNA using mechanical disruption combined with the guanidine-based reagent DNAzol. Template DNA was isolated by this method from cultures of Campylobacter jejuni resistant to lysis by boiling or enzymes and identified following polymerase chain reaction (PCR) amplification using primers specific for the hippuricase gene. Direct detection of campylobacters in poultry-processing samples by PCR is demonstrated in chicken carcass rinses spiked with lysis-resistant C. jejuni. Our results indicate that this method of DNA isolation may be ideal for direct PCR detection of pathogenic bacteria in complex samples of widely varied origin, especially when the target organisms are difficult to lyse by other means.

Animals↗
PubMed2000

An evaluation of methods for the detection of spinal cord in product derived from advanced meat recovery systems.

This study investigated the usefulness of immunohistochemistry, histochemistry, and polarization microscopy in documenting the presence of spinal cord in ground product produced by advanced meat recovery systems (AMRS). A battery of diagnostic stains proved most helpful for definitive documentation of spinal cord in the AMRS product. Antibodies to neurofilament and glial fibrillary acidic proteins were useful for highlighting the presence of neural tissue in comminuted product but when used alone did not definitively differentiate central nervous system tissue from peripheral nerve. Polariscopy, trichrome staining, and immunostaining for synaptophysin were very useful for differentiating spinal cord from peripheral nerve. Spinal cord was found in 2 of 196 AMRS samples when only hematoxylin and eosin staining was used. In a later immunohistochemical subset of the survey study, spinal cord was found in 7 of 17 AMRS samples. More recently, spinal cord tissue has been found in 3 of 26 regulatory samples using hematoxylin and eosin stains combined with immunohistochemical techniques.

Animals↗
PubMed1999

Detection of multidrug-resistant Salmonella enterica serotype typhimurium DT104 based on a gene which confers cross-resistance to florfenicol and chloramphenicol.

Salmonella enterica serotype typhimurium (S. typhimurium) DT104 (DT104) first emerged as a major pathogen in Europe and is characterized by its pentadrug-resistant pattern. It has also been associated with outbreaks in the United States. The organism typically carries resistance to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline. The mechanism of chloramphenicol resistance in DT104 was determined by producing antibiotic-resistant Escherichia coli host strain clones from DT104 DNA. DNA from chloramphenicol-resistant clones was sequenced, and probes specific for the genes floS. typhimurium (floSt), int, invA, and spvC were produced for colony blot hybridizations. One hundred nine Salmonella isolates, including 44 multidrug-resistant DT104 isolates, were tested to evaluate the specificities of the probes. The gene floSt, reported in this study, confers chloramphenicol and florfenicol resistance on S. typhimurium DT104. Florfenicol resistance is unique to S. typhimurium DT104 and multidrug-resistant S. typhimurium isolates with the same drug resistance profile among all isolates evaluated. Of 44 DT104 isolates tested, 98% were detected based on phenotypic florfenicol resistance and 100% had the floSt-positive genotype. Resistances to florfenicol and chloramphenicol are conferred by the gene floSt, described in this paper. Presumptive identification of S. typhimurium DT104 can be made rapidly based on the presence of the floSt gene or its resulting phenotype.

Animals↗
PubMed1998

Regression of subcutaneous lymphoma following removal of an ovarian granulosatheca cell tumor in a horse.

A 9-year-old Arabian mare was admitted for evaluation of multiple subcutaneous nodules and infertility. Fine-needle aspiration of one of the subcutaneous nodules resulted in a cytologic diagnosis of histiolymphocytic lymphoma. Palpation per rectum and transrectal ultrasonography revealed a mass associated with the left ovary. Excision of the ovarian tumor was performed, and a histopathologic diagnosis of granulosa-theca cell tumor was made. After removal of the granulosa-theca cell tumor, subcutaneous nodules regressed. The referring veterinarian reported that the nodules had also disappeared and then recurred after administration of a synthetic progestin. To further characterize the lymphoma and investigate this possible hormonal relationship, immunophenotyping and estrogen and progesterone receptor assays were performed. The subcutaneous lymphoma was classified as a T-cell rich B-cell lymphoma, results of estrogen receptor assays were negative, and results of progesterone receptor assays were positive. Clinical observations of subcutaneous lymphoma in horses indicate that the waxing and waning nature of these tumors may be associated with the estrous cycle, pregnancy, foaling, and lactation. Clinical observations and identification of progesterone receptors suggest that a relationship between serum steroid hormone concentrations, such as estrogen and progesterone, and subcutaneous lymphoma may exists.

Animals↗
PubMed1998

Equine malignant lymphomas: morphologic and immunohistochemical classification.

Gross lesions, microscopic appearance, and immunophenotyping are reported in a retrospective study of 31 cases of equine malignant lymphoma. Immunohistochemical studies were performed on archived formalin-fixed, paraffin-embedded tissues. Monoclonal antibodies to surface glycoprotein BLA.36 and intracytoplasmic domains of mb-1 and B29 were used to document the presence of B lymphocytes in the equine tumors. Polyclonal antibody to CD3 and monoclonal antibodies to T-lymphocyte markers CD3 and CD5 revealed the presence of variable numbers of T cells within the equine lymphomas. The neoplastic component of the equine lymphomas was determined through morphologic evaluation, immunophenotyping, and the use of proliferation markers Ki-67 and proliferating cell nuclear antigen. Equine malignant lymphomas were composed of a heterogeneous cell population. Most tumors contained B and T lymphocytes. Twenty-four horses had diffuse lymphomas derived from B lymphocytes. Thirteen of these lymphomas contained primarily neoplastic B lymphocytes. Eleven additional cases of diffuse large B-cell lymphoma contained from 40% to 80% nonneoplastic T lymphocytes and were classified as T-cell-rich, large B-cell lymphomas. This is the first description of T-cell-rich, B-cell lymphoma in the horse. Six tumors with a diffuse architecture were derived from T lymphocytes. Four T-cell tumors were large-cell tumors, 1 was a small-cell tumor, and in 1 tumor the size of the cells could not be determined accurately because of autolytic change in the tissues. One diffuse large-cell lymphoma did not react with either B- or T-cell markers.

Animals↗
PubMed1997

Detection of equine and bovine T- and B-lymphocytes in formalin-fixed paraffin-embedded tissues.

Formalin-fixed paraffin-embedded sections of equine and bovine lymph nodes, spleen, thymus, and Peyer's patches were incubated with monoclonal antibodies to B-lymphocyte markers BLA.36, B29, and mb-1 and T-lymphocyte markers CD3 and CD5. The monoclonal antibody BLA.36 reacted with 80-90% of lymphocytes in the germinal centers and mantle zones of follicles in lymph nodes, spleen, and Peyer's patches. In addition, 90% of lymphocytes in the marginal zone of the spleen, and variable numbers of lymphocytes within lymph node medullary cords were immunopositive for BLA.36. Antibodies to B29 and mb-1 produced similar staining patterns as BLA.36 with fewer positive cells in the germinal centers and medullary cords. BLA.36, B29, and mb-1 reacted with 30-50% of lymphocytes in the medulla of the thymus and with 5-10% of lymphocytes in the cortex. CD3 and CD5 reacted with 90% of lymphocytes in the paracortex and parafollicular zones of lymph nodes, spleen, and Peyer's patches; 40-50% of lymphocytes in the medullary cords of lymph nodes, and scattered positive cells within follicles. Anti-CD3 antibody reacted with 95% of lymphocytes in the splenic red pulp, but antibodies directed against CD5 reacted only faintly with approximately 5-10% of lymphocytes in the red pulp. CD3 and CD5 reacted with 50-60% of cells in the medulla of the thymus and with 40-80% of lymphocytes in the thymic cortex. The biochemical characterization of the antibodies by Western blotting against lysates of equine and bovine peripheral blood mononuclear cells confirmed that antibodies to BLA.36, mb-1, B29, CD3, and CD5 detected molecules of the same approximate molecular mass as found on lymphoid cells of human beings and rats.

Animals↗
PubMed1996

A retrospective study of multicentric bovine renal cell tumors.

Tissues from twenty mature cows with primary renal cell tumors were submitted over an 11-year period because of gross lesions detected during routine slaughter and inspection. Tumors visualized grossly were multiple and bilateral in seven cattle, multiple within one kidney in four cattle, and solitary in nine cattle. The tumors were primarily cortical, yellow-orange to tan, proliferative, well circumscribed, and extended above the capsular surface of the kidney. Tumors were microscopically multiple even when grossly described as solitary lesions, except in one cow. Twelve tumors (60%) were microscopically multiple in one kidney, seven tumors (35%) were multiple and bilateral, and only one cow (5%) exhibited extrarenal metastasis. Tumors from nineteen cows were composed of eosinophilic granular epithelial cells; tumors from one cow were clear cell type. Each tumor contained several histologic patterns. Corpora amylacea, proteinaceous secretions, and hemosiderin were characteristic findings in bovine renal cell carcinoma. All 20 cows with renal cell tumors exhibited positive immunoreactivity to uromodulin (Tamm-Horsfall protein).

Animals↗
PubMed1996

A retrospective study of pancreatic tumors in slaughter cattle.

Sixteen primary pancreatic tumors were found in a retrospective study of bovine pancreatic lesions detected in slaughtered cattle. Eleven islet cell tumors and three pancreatic exocrine carcinomas were identified based on light microscopy and immunohistochemistry. Nine of 11 islet cell tumors were classified as malignant. Metastatic sites included iliac, mediastinal, hepatic, and mesenteric lymph nodes, peritoneum, mesentery, and liver. Six cows with multiple islet cell tumors also had pheochromocytomas. All 11 islet cell tumors had positive immunoreactivity to insulin and somatostatin. Three tumors also contained cells immunoreactive for glucagon and two tumors contained pancreatic polypeptide immunoreactive cells. Immunoreactivity of tumor cells in metastatic sites was similar to their respective primary tumors. All exocrine pancreatic carcinomas metastasized widely and were immunonegative for insulin, somatostatin, glucagon, and pancreatic polypeptide. No mixed endocrine-exocrine tumors were identified. None of the endocrine or exocrine tumors contained amyloid. Additional primary tumors of the bovine pancreas included one neurofibroma and one neurofibrosarcoma. Additional cases with lesions of the bovine pancreas included nodular hyperplasia in 15 cows, exocrine acinar atrophy and fibrosis in four cows (two of which also had pancreatic lithiasis), pancreatitis in one cow, peripancreatic fibrosis in two cows, pancreatic steatosis in one animal, and pancreatic hemorrhages in one cow.

Adenoma, Islet Cell↗
PubMed1995

Alternate routes of invasion may affect pathogenesis of Salmonella typhimurium in swine.

Transmission of Salmonella typhimurium in swine is traditionally believed to occur by the fecal-oral route, with invasion through the intestinal wall and Peyer's patches. However, involvement of the upper respiratory tract may be equally important. An esophagotomy was performed on 6- to 8-week-old pigs. Esophagotomized pigs were challenged intranasally with 10(9) CFU of S. typhimurium cells and necropsied at 3, 6, 12, and 18 h postinoculation (p.i.). By 3 h p.i., S. typhimurium was recovered from cecum, colon, head, and thoracic tissues and from the middle ileum involving a large number of Peyer's patches. The ileocolic lymph nodes and ileocolic junction were not positive for S. typhimurium until 6 and 12 h p.i., respectively. Additional pigs were inoculated transthoracically with 10(9) CFU of S. typhimurium and necropsied at 3 and 18 h p.i. By 3 h p.i., all tissues were positive for S. typhimurium. Tonsil explants seeded with 10(9) CFU of S. typhimurium indicated that within 6 h p.i., S. typhimurium was located within the tonsilar crypts. These data show that after intranasal inoculation, S. typhimurium rapidly appears in the gut tissues and suggest that the tonsils and lung may be important sites for invasion and dissemination of Salmonella species.

Animals↗
PubMed1995

Spontaneous equine pulmonary granular cell tumors: morphologic, histochemical, and immunohistochemical characterization.

Spontaneous equine pulmonary granular cell tumors were diagnosed in six mature horses at slaughter. These tumors were grossly recognized as multiple (5/6) or single (1/6) creamy white, firm nodules. The tumors, located adjacent to bronchi and bronchioles, often invaded airways, resulting in partial to complete occlusion of the lumina. Neoplastic cells were rounded to polyhedral with numerous eosinophilic cytoplasmic granules that reacted uniformly positive with S-100 and neuron-specific enolase antibodies and multifocally with glial fibrillary acidic protein antibodies. These cells were negative for muscle-specific actin, lysozyme, cytokeratin, chromogranin A, and myelin basic protein antigens and did not stain with silver by the Grimelius technique. Uniformly blue-green and scattered pink intracytoplasmic granules were evident with luxol fast blue and periodic acid-Schiff counterstain for myelin and myelin breakdown products. Histochemical and immunohistochemical staining results of these tumors suggest that they are composed primarily of myelinating Schwann cells with lesser numbers of scattered nonmyelinating Schwann cells. The morphologic features of the equine pulmonary granular cell tumors are strikingly similar to those of endobronchial granular cell tumors of human beings.

Animals↗
PubMed1995

Malignant melanocytic neural crest tumor in a young chicken (Gallus domesticus).

A malignant melanocytic tumor was found in an 8-week-old chicken. The tumor, which was composed of melanocytes, ganglion cells, nerves, and primitive pressure receptors, was examined by light microscopy, transmission electron microscopy, and immunohistochemistry. Antibodies to neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP), and S-100 protein variably stained nerves, but melanocytes were only rarely immunolabelled by NSE antibodies and there was no specific staining of these cells for S-100 or GFAP. Ultrastructurally, neoplastic melanocytes contained melanin within melanosomes and premelanosomes and did not resemble Schwann cells.

Adrenal Gland Neoplasms↗
PubMed1995

Icterus in bob veal calves and its association with lack of colostrum intake and high serum creatine kinase activity.

Icterus condemnations compose a substantial proportion (41%) of total condemnations of bob veal, the class of veal composed of calves < 3 weeks old and weighing up to 68 kg. At postmortem examination, bob veal condemned because of icterus have generalized yellow discoloration of tissues, which is commonly associated with large, yellow liver (fatty liver), and a paucity of other gross pathologic changes. To establish that the generalized yellow discoloration was attributable to high tissue bilirubin concentrations and to examine the underlying mechanism(s) that might be responsible, blood samples and tissue specimens were obtained from clinically normal and icteric bob veal calves at slaughter. For comparison, blood samples were collected from clinically normal, 1- to 5-day-old Holstein calves being raised on local dairy farms. Hematologic and serum biochemical analyses were obtained for the 3 groups of calves (normal local, normal slaughter, and icteric slaughter), and tissues of slaughter calves were examined for histologic evidence of inflammatory or degenerative changes. Mean +/- SD total bilirubin concentration and creatine kinase (CK) activity in icteric bob veal (3.3 +/- 0.8 mg/dl; 869 +/- 788 U/L), normal bob veal (1.4 +/- 0.7 mg/dl; 486 +/- 890 U/L), and normal local calves (0.5 +/- 0.2 mg/dl; 156 +/- 158 U/L) were significantly different. When data for both normal and icteric bob veal calf groups were combined for analysis, total bilirubin concentration regressed significantly on hepatic lipid scores (P = 0.00003) and CK activity (P = 0.00049). Colostrum consumption was determined by measuring serum total protein concentration and serum gamma-glutamyltransferase activity. Bob veal calves that had not consumed colostrum had significantly higher total bilirubin (P = 0.00005) and CK (P = 0.0008) values. It was concluded that normal and icteric bob veal calves have significant increase in total bilirubin concentration, and icterus of bob veal calves is secondary to unconjugated hyperbilirubinemia. Lack of colostrum consumption was strongly correlated with icterus in bob veal calves.

Analysis of Variance↗
PubMed1994

Bovine pulmonary blastomas: histomorphologic description and immunohistochemistry.

Three adult female dairy cattle with pulmonary blastomas were evaluated. The gross lesions at postmortem were described as multiple white circumscribed masses throughout the lungs, with pleural involvement in one cow and lymph node metastasis in the other two cows. Histologically, the tumors contained a dual population of mesenchymal and epithelial neoplastic cells. Epithelial cells formed nests, tubules, and formations resembling bronchioles of normal lung. Mesenchymal cells were spindle shaped with oval nuclei and fibrillar eosinophilic cytoplasm, were large rounded cells with multiple round nuclei and granular eosinophilic cytoplasm, or appeared blast-like, with large hyperchromatic nuclei and amphophilic cytoplasm. The tumors varied greatly in appearance from one field to another within the same tumor. Epithelial cells stained positively with anticytokeratin antibodies. Some spindle-shaped mesenchymal cells exhibited smooth muscle cell differentiation with positive staining with anti-vimentin, anti-muscle-specific actin, and anti-smooth muscle actin antibodies, whereas other rounded mesenchymal cells expressed striated muscle cell differentiation with multiple nuclei and positivity to anti-neuron-specific enolase and anti-muscle-specific actin antibodies. The variable expression of the intermediate filaments and cytoplasmic enzymes indicates multiple pathways of differentiation in the pulmonary blastomas.

Animals↗