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Biomedical subjects

K M Young

Publications and source records attributed to K M Young.

At least 19 recordsLinked to original sources
PubMed2006

SOCS3 negatively regulates LIF signaling in neural precursor cells.

Cytokines that signal through the LIFRbeta/gp130 receptor complex, including LIF and CNTF, promote the self-renewal of embryonic and adult neural precursor cells (NPCs). In non-CNS tissues, the protein suppressor of cytokine signaling-3 (SOCS3) negatively regulates signaling through gp130. Here, we analyze the role of SOCS3 in inhibiting LIF signaling in NPCs in vitro. SOCS3 is rapidly expressed by NPCs in response to LIF stimulation, with this expression largely dependent on recruitment of STAT proteins to the activated gp130 receptor. Proliferating NPC cultures can be generated from SOCS3 knockout (SOCS3KO/KO) embryos and display prolonged STAT3 phosphorylation and induction of the GFAP gene in response to LIF. In comparison with SOCS3 wild-type (SOCS3WT/WT) NPCs, SOCS3KO/KO cultures display enhanced self-renewal capacity. However, the clonal potential of SOCS3WT/WT but not SOCS3KO/KO NPCs is enhanced by exogenous LIF. Thus, SOCS3 acts as a negative regulator of LIF signaling in NPCs.

Animals↗
PubMed2004

Noninvasive monitoring of adrenocortical activity in carnivores by fecal glucocorticoid analyses.

Measurement of glucocorticoid metabolites in feces has become an accepted method for the noninvasive evaluation of adrenocortical activity. The objective of this study was to determine if a simple cortisol enzyme immunoassay (EIA) was suitable for monitoring adrenocortical activity in a variety of carnivore species. Performance of the cortisol EIA was gauged by comparison to a corticosterone radioimmunoassay (RIA) that has been used for measuring glucocorticoid metabolites in feces of numerous species. Tests for parallelism and extraction efficiency were used to compare the cortisol EIA and corticosterone RIA across eight species of carnivores (Himalayan black bear, sloth bear, domestic cat, cheetah, clouded leopard, black-footed ferret, slender-tailed meerkat, and red wolf). The biological relevance of immunoreactive glucocorticoid metabolites in feces was established for at least one species of each Carnivora family studied with an adrenocorticotropic hormone (ACTH) challenge. High performance liquid chromatography (HPLC) analysis of fecal extracts for each species revealed (1) the presence of multiple immunoreactive glucocorticoid metabolites in feces, but (2) the two immunoassays measured different metabolites, and (3) there were differences across species in the number and polarities of metabolites identified between assay systems. ACTH challenge studies revealed increases in fecal metabolite concentrations measured by the cortisol EIA and corticosterone RIA of approximately 228-1145% and approximately 231-4150% above pre-treatment baseline, respectively, within 1-2 days of injection. Concentrations of fecal glucocorticoid metabolites measured by the cortisol EIA and corticosterone RIA during longitudinal evaluation (i.e., >50 days) of several species were significantly correlated (P<0.0025, correlation coefficient range 0.383-0.975). Adrenocortical responses to physical and psychological stressors during longitudinal evaluations varied with the type of stimulus, between episodes of the same stimulus, and among species. Significant elevations of glucocorticoid metabolites were observed following some potentially stressful situations [anesthesia (2 of 3 subjects), restraint and saline injection (2 of 2 subjects), restraint and blood sampling (2 of 6 episodes), medical treatment (1 of 1 subject)], but not in all cases [e.g., gonadotropin injection (n=4), physical restraint only (n=1), mate introduction/breeding (n=1), social tension (n=1), construction (n=2) or relocation (n=1)]. Results reinforced the importance of an adequate baseline period of fecal sampling and frequent collections to assess adrenocortical status. The corticosterone RIA detected greater adrenocortical responses to exogenous ACTH and stressful exogenous stimuli in the Himalayan black bear, domestic cat (female), cheetah, clouded leopard, slender-tailed meerkat, and red wolf, whereas the cortisol EIA proved superior to resolving adrenocortical responses in the black-footed ferret and domestic cat (male). Overall results suggest the cortisol EIA tested in this study offers a practical method for laboratories restricted in the usage of radioisotopes (e.g., zoological institutions and field facilities) to integrate noninvasive monitoring of adrenocortical activity into studies of carnivore behavior and physiology.

Acinonyx↗
PubMed2002

Recovery of ischaemic injured porcine ileum: evidence for a contributory role of COX-1 and COX-2.

BACKGROUND: We have previously shown that the non-selective cyclooxygenase (COX) inhibitor indomethacin retards recovery of intestinal barrier function in ischaemic injured porcine ileum. However, the relative role of COX-1 and COX-2 elaborated prostaglandins in this process is unclear. AIMS: To assess the role of COX-1 and COX-2 elaborated prostaglandins in the recovery of intestinal barrier function by evaluating the effects of selective COX-1 and COX-2 inhibitors on mucosal recovery and eicosanoid production. METHODS: Porcine ileal mucosa subjected to 45 minutes of ischaemia was mounted in Ussing chambers, and transepithelial electrical resistance was used as an indicator of mucosal recovery. Prostaglandins E1 and E2 (PGE) and 6-keto-PGF1alpha (the stable metabolite of prostaglandin I2 (PGI2)) were measured using ELISA. Thromboxane B2 (TXB2, the stable metabolite of TXA2) was measured as a likely indicator of COX-1 activity. RESULTS: Ischaemic injured tissues recovered to control levels of resistance within three hours whereas tissues treated with indomethacin (5x10(-6) M) failed to fully recover, associated with inhibition of eicosanoid production. Injured tissues treated with the selective COX-1 inhibitor SC-560 (5x10(-6) M) or the COX-2 inhibitor NS-398 (5x10(-6) M) recovered to control levels of resistance within three hours, associated with significant elevations of PGE and 6-keto-PGF1alpha compared with untreated tissues. However, SC-560 significantly inhibited TXB2 production whereas NS-398 had no effect on this eicosanoid, indicating differential actions of these inhibitors related to their COX selectivity. CONCLUSIONS: The results suggest that recovery of resistance is triggered by PGE and PGI2, which may be elaborated by either COX-1 or COX-2.

Animals↗
PubMed2001

Overview and assessment of the suicidal child.

A comprehensive evaluation and assessment of the suicidal child is important to target those at risk. This article identifies risk factors for suicide in children and adolescents and provides a step-by-step interview scheme that captures all relevant information that is obtained when assessing these children and adolescents.

Adolescent↗
PubMed2001

PGE2 triggers recovery of transmucosal resistance via EP receptor cross talk in porcine ischemia-injured ileum.

16,16-Dimethyl-PGE2 (PGE2) may interact with one of four prostaglandin type E (EP) receptors, which signal via cAMP (via EP2 or EP4 receptors) or intracellular Ca(2+) (via EP1 receptors). Furthermore, EP3 receptors have several splice variants, which may signal via cAMP or intracellular Ca(2+). We sought to determine the PGE2 receptor interactions that mediate recovery of transmucosal resistance (R) in ischemia-injured porcine ileum. Porcine ileum was subjected to 45 min of ischemia, after which the mucosa was mounted in Ussing chambers. Tissues were pretreated with indomethacin (5 microM). Treatment with the EP1, EP2, EP3, and EP4 agonist PGE2 (1 microM) elevated R twofold and significantly increased tissue cAMP content, whereas the EP2 and EP4 agonist deoxy-PGE1 (1 microM) or the EP1 and EP3 agonist sulprostone (1 microM) had no effect. However, a combination of deoxy-PGE1 and sulprostone stimulated synergistic elevations in R and tissue cAMP content. Furthermore, treatment of tissues with deoxy-PGE1 and the Ca(2+) ionophore A-23187 stimulated synergistic increases in R and cAMP, indicating that PGE2 triggers recovery of R via EP receptor cross talk mechanisms involving cAMP and intracellular Ca(2+).

Animals↗
PubMed2000

Immunodiagnostic testing in horses.

Technologic advances in immunodiagnostic testing have enhanced our understanding of the pathogenesis of a broad array of diseases, including infectious diseases, immunodeficiency disorders, and immune-mediated disorders. If applied rationally, with an understanding of the questions the tests answer as well as the limitations that constrain their use, these tests can serve as valuable aids in the diagnosis and management of equine diseases.

Animals↗
PubMed2000

Genistein augments prostaglandin-induced recovery of barrier function in ischemia-injured porcine ileum.

We have previously shown that PGE(2) enhances recovery of transmucosal resistance (R) in ischemia-injured porcine ileum via a mechanism involving chloride secretion. Because the tyrosine kinase inhibitor genistein amplifies cAMP-induced Cl(-) secretion, we postulated that genistein would augment PGE(2)-induced recovery of R. Porcine ileum subjected to 45 min of ischemia was mounted in Ussing chambers, and R and mucosal-to-serosal fluxes of [(3)H]N-formyl-methionyl-leucyl phenylalanine (FMLP) and [(3)H]mannitol were monitored as indicators of recovery of barrier function. Treatment with genistein (10(-4) M) and PGE(2) (10(-6) M) resulted in synergistic elevations in R and additive reductions in mucosal-to-serosal fluxes of [(3)H]FMLP and [(3)H]mannitol, whereas treatment with genistein alone had no effect. Treatment of injured tissues with genistein and either 8-bromo-cAMP (10(-4) M) or cGMP (10(-4) M) resulted in synergistic increases in R. However, treatment of tissues with genistein and the protein kinase C (PKC) agonist phorbol myristate acetate (10(-5)-10(-6) M) had no effect on R. Genistein augments recovery of R in the presence of cAMP or cGMP but not in the presence of PKC agonists.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1999

Usefulness of aerobic microbial culture and cytologic evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis in animals.

OBJECTIVE: To determine the diagnostic value of aerobic microbial culture and cytologic evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis (IUK). DESIGN: Prospective study. ANIMALS: 48 animals (26 dogs, 13 horses, 7 cats, 1 bird, and 1 llama) with corneal ulcers. PROCEDURE: Scrapings from corneal ulcers were examined cytologically. Corneal swab specimens were submitted for microbial culture. Animals were grouped according to whether they had been receiving antimicrobials at the time of admission. RESULTS: Of the 38 animals receiving antimicrobials, 19 had positive results for IUK on cytologic evaluation, 20 on microbial culture, and 26 on cytologic evaluation, microbial culture, or both. Of the 10 animals not receiving antimicrobials at the time of admission, 7 had positive results for IUK on cytologic evaluation, and 9 had positive results on microbial culture. In this group of 10 animals, additional animals with IUK were not identified on the basis of cytologic evaluation alone. When all 48 animals were considered irrespective of antimicrobial treatment, 26 and 29 had positive results for IUK on cytologic evaluation and microbial culture, respectively, whereas IUK was confirmed in 35 animals on the basis of cytologic evaluation, microbial culture results, or both. CONCLUSIONS AND CLINICAL RELEVANCE: Microbial culture and cytologic evaluation of corneal specimens maximizes identification of IUK, especially in animals receiving antimicrobial treatment. Because of serious consequences of untreated IUK, we recommend that both diagnostic tests be used to tailor treatment and reduce risk of vision impairment in animals.

Animal Diseases↗
PubMed1999

Change in stress outcomes following an industrial dispute in the ambulance service: a longitudinal study.

A longitudinal study to assess change in the symptoms of strain following an industrial dispute was carried out in a North of England ambulance service. A cohort of ambulance staff was surveyed over three stages: during the dispute; during the process of reorganizing the service; and during the implementation of a new operating structure. Dependent variables included measure of job satisfaction and mental and physical health. Independent variables included measures of the type A behaviour pattern, locus of control, source of perceived pressure and coping techniques. The results failed to support the main hypothesis as the symptoms of strain measured did not show any reduction after the dispute. Job dissatisfaction increased in the aftermath of the dispute and remained poor during the implementation of change. Symptoms of mental and physical ill-health showed no reduction following the dispute but symptoms of mental ill-health increased as organizational change was implemented. The high level of 'externality' shown by ambulance staff partially accounted for both these increases. It is proposed that the manner in which the aftermath of the dispute and ensuing organizational change were managed gave an increasingly limited scope for influence.

Adaptation, Psychological↗
PubMed1997

Occupational stress in the ambulance service: a diagnostic study.

The Occupational Stress Indicator (OSI) was used to investigate job stress in an ambulance service in the northwest of England. Seven different aspects of the stress-strain relationship were assessed and the findings compared with those from the fire service. Ambulance service employees were found to be experiencing major stress outcomes of low job satisfaction and poor mental and physical health. Fire service employees revealed significantly poorer physical health. Assesses the sources of job stress, type A behaviour, locus of control and coping styles and discusses them in the light of change in the public services.

Adaptation, Psychological↗
PubMed1997

Characterization of eosinophil progenitor cells in feline bone marrow.

OBJECTIVE: To identify eosinophil progenitor cells in feline bone marrow, establishing an assay method to use in studies of eosinophilopoiesis and eosinophilopoietic factors in cats. ANIMALS: Healthy, laboratory animal source cats. PROCEDURE: Sources of colony-stimulating activity were prepared by conditioning media with bone marrow, spleen, and blood mononuclear cells from cats infected with Toxocara canis. Bone marrow cells were aspirated and cultured to develop the eosinophil progenitor cell assay and to test cells from 9 healthy cats in the assay. RESULTS: Optimal conditions for identifying colony-forming units-eosinophil and cluster-forming units-eosinophil were as follows. Bone marrow mononuclear cells (10(5)) were plated in 1 ml of supplemented medium, fetal bovine serum, and agar. The source of eosinophil growth factor(s) was bone marrow-conditioned medium made in the presence of 2.5 micrograms of concanavalin A/ml; other conditioned media also supported eosinophil colony growth. Dishes were incubated for 7 days at 37 C and 7% CO2. The colony-forming units-eosinophil formed aggregates of > 50 Luxol fast blue-positive cells and had dispersed morphology; the cluster-forming units-eosinophil formed aggregates of < 50 cells. CONCLUSION AND CLINICAL RELEVANCE: Similar to other species, cats have separate and distinct eosinophil progenitor cells. The eosinophil progenitor assay may be used to characterize altered kinetics of eosinophilopoiesis, to assess eosinophil growth factors, and to evaluate therapeutic regimens that might be useful in the management of excess eosinophil production.

Animals↗
PubMed1996

Sex differences in rat hepatic cytolethality of the protein kinase C inhibitor safingol: role of biotransformation.

Safingol [(2S,3S)-2-amino-1,3-octadecanediol], a sphingosine analog that inhibits protein kinase C, was developed to treat dermatoses and cancer. Preclinical toxicology studies performed to assess the effects of safingol showed that 6 weeks of dermal application over 10% of body surface area caused dose-dependent increases in serum enzymes and hepatic histopathological changes associated with liver damage in female rats. Liver toxicity was not seen in male rats at the same doses. Plasma safingol concentrations were similar in male and female rats following topical exposure. The underlying mechanism(s) for the sex differences in toxicity in rats were examined using isolated hepatocytes. An in vitro model of male versus female differences in safingol.HCl-induced hepatotoxicity was established using a suspension/culture technique. Concentrations of safingol.HCl which produced cytolethality in 50% of the hepatocytes were 125 and 48 microM for male and female rat hepatocytes, respectively. Cytolethality was time-, concentration-, and cell number-dependent. Inhibition of cytochrome P450 in vitro with 1-phenylimidazole increased safingol.HCl-induced cytolethality in male but not female hepatocytes, suggesting that male rat hepatocytes have a cytochrome p450 isoenzyme which metabolizes safingol.HCl to an inactive metabolite thus reducing hepatotoxicity. Furthermore, in vivo pretreatment with the CYP4A-inducing agent, clofibrate, protected both male and female hepatocytes from cytolethality. The results of this study indicate that the sex differences seen in hepatotoxicity could be due to differences in biotransformation such that female rat hepatocytes either lack or have a reduced constitutive level of a cytochrome P450 isoenzyme that metabolizes safingol to a nontoxic metabolite. In addition, safingol produced hepatocyte cell death without inflammation in vivo, and a "ladder-like" DNA fragmentation pattern in vitro, consistent with an apoptotic mechanism of cell death.

Administration, Oral↗
PubMed1994

Identification and characterization of the human cell adhesion molecule contactin.

We have prepared a monoclonal antibody, Neuro-1, that recognizes the human homolog of the chicken contactin/F11 and mouse F3 cell adhesion molecules. The Neuro-1 antigen, structurally characterized as a 135 kDa glycosylphosphatidylinositol-linked glycoprotein, was immunoaffinity purified and partially sequenced. Comparison of an internal peptide sequence to that predicted from the chicken contactin/F11, mouse F3 and human contactin (reported herein) cDNA sequence identifies the Neuro-1 antigen as human contactin. Moreover, a polyclonal antisera generated against the purified Neuro-1 antigen was immunoreactive with a fragment of human contactin expressed in bacteria. The complete coding and deduced amino acid sequences of human contactin were determined and are 86% and 95% identical to the respective mouse F3 sequences. Structural features shared with contactin/F11/F3 include six immunoglobulin type C2 and four fibronectin type III-like domains, multiple sites for asn-linked glycosylation and a COOH-terminal signal peptide presumably removed during the generation of a phosphatidylinositol cell surface linkage. The potential for glycosylation and GPI-linkage is also consistent with protein chemical studies of human contactin. Contactin mRNA expression was characterized using Northern blot analyses of human tissues and cell lines. High level expression of a single contactin transcript in adult brain, and low level expression of multiple transcripts in lung, pancreas, kidney and skeletal muscle are observed. Highly expressed multiple transcripts, similar in pattern to that of pancreas, lung, kidney and skeletal muscle, are also observed in human neuroblastoma and retinoblastoma cell lines.

Amino Acid Sequence↗
PubMed1993

Deprotection of methylphosphonate oligonucleotides using a novel one-pot procedure.

Deprotection of methylphosphonate oligonucleotides with ethylenediamine was evaluated in a model system. Methylphosphonate sequences of the form 5'-TTTNNTTT, where N was either N4-bz-dC, N4-ibu-dC, N2-ibu-O6-DPC-dG, N2-ibu-dG, N6-bz-dA, or T, were used to determine the extent of modifications that occur during deprotection. Up to 15% of N4-bz-dC was found to transaminate at the C4 position when treated with ethylenediamine. A similar displacement reaction with ethylenediamine was observed at the O6 position of N2-ibu-O6-DPC-dG, and to a much lesser extent of N2-ibu-dG. Side reactions were not observed when oligonucleotides containing N4-ibu-dC, N6-bz-dA, or T were treated with ethylenediamine. A novel method of deprotecting methylphosphonate oligonucleotides was developed from these studies. The method incorporates a brief treatment with dilute ammonia for 30 minutes followed by addition of ethylenediamine for 6 hours at room temperature to complete deprotection in a one-pot format. The solution is then diluted and neutralized to stop the reaction and prepare the crude product for chromatographic purification. This method was used to successfully deprotect a series of oligonucleotides at the 1, 100, and 150 mumole scales. These deprotection results were compared to a commonly used two-step method and found to be superior in yield of product by as much as 250%.

Ammonia↗
PubMed1993

Primary mesenchymal or mixed-cell-origin lung tumors in four dogs.

Primary lung tumors of mesenchymal or mixed cell origin were diagnosed in 4 dogs with clinical and radiographic abnormalities indicating an intrathoracic mass. Each dog had 1 large intrapulmonary lesion, and 1 dog also had nodules scattered throughout all lung lobes. Two dogs were euthanatized; 1 had a biphasic pulmonary blastoma; and the other had a pulmonary chondroblastic osteosarcoma with intrapulmonary metastases. The masses in the other 2 dogs were hamartomas (lipomatous in 1, microcystic in the other), which were resected. Both dogs survived more than 1 year after surgery. Primary lung tumors are uncommon in dogs and are generally malignant (adenocarcinomas or carcinomas). Tumors of connective tissue or mixed cell origin are rare, but the outcome is potentially good after surgical removal.

Animals↗
PubMed1993

An improved method for the synthesis and deprotection of methylphosphonate oligonucleotides.

The methylphosphonate oligonucleotide synthesis methods described here give the desired products in good yield. Superior amounts of product are achieved by modifying both the DNA synthesis program and the reagent to compensate for the unstable methylphosphonite intermediate. Deprotection conditions have also been altered to maximize the recovery of oligonucleotide from DNA synthesis supports and to minimize the amount of base modification. Mass-spectrometry analysis of our oligonucleotides has verified their purity and confirmed the absence of modified bases. When compared to standard DNA synthesis methods, this procedure uses only about one-third the usual amount of monomer. Using these procedures, it should be possible to synthesize reliably methylphosphonate oligonucleotides at 1- and 15-mumol scales.

Base Sequence↗