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Biomedical subjects

K Hunter

Publications and source records attributed to K Hunter.

At least 55 records · Page 3Linked to original sources

Assessment of skeletal muscle viability by PET.

UNLABELLED: We investigated the use of [18F]fluoro-2-deoxyglucose (FDG) PET scanning for assessment of skeletal muscle viability in patients with peripheral vascular disease and in patients following free-flap skeletal muscle transfer for closure of open wounds. METHODS: We obtained 32 FDG-PET scans from 30 patients, either at the time of admission for peripheral vascular disease (n = 16) or between 1 and 15 days after surgery for skeletal muscle transfer (n = 16). Ratios between injured and contralateral limb FDG tracer activity uptake were correlated with clinical outcome at 1 mo to 3 yr follow-up. RESULTS: Viable muscle uptake ratios ranged from 0.47 to 7.88 (mean: 2.26 +/- 1.81; n = 26), while nonviable muscle uptake ratios ranged from 0.12 to 0.46 (mean: 0.27 +/- 0.12; n = 6; p < 0.02). After skeletal muscle transfer, two patients with viable tissue, as documented by PET, required amputation due to osteomyelitis, and one patient with peripheral vascular disease who showed viable tissue by PET required amputation 3 mo after the PET scan because of recurrent ulcers. CONCLUSION: FDG-PET scanning can determine skeletal muscle viability in patients with peripheral vascular disease and in patients following free-flap transfer.

Adolescent↗

IRE-bubble PCR: a rapid method for efficient and representative amplification of human genomic DNA sequences from complex sources.

A significant issue in the analysis of any genomic DNA segment is the generation of a unique set of short single-copy sequences that are representative of that region. In this report we describe a novel technique, IRE-bubble PCR, which was designed to amplify the human DNA content of somatic cell hybrids, YACs, cosmids, and lambda phage and result in greater complexity and representation than standard inter-IRE PCR. Here we demonstrate that IRE-bubble PCR is species specific and that it results in the generation of a product that is at least 10-fold more complex and representative than that produced by standard inter-IRE PCR. In addition, we have addressed the factors that contribute to the representation of the IRE-bubble PCR product and show how they may be used to further increase the complexity of this reaction. Finally, we have illustrated how the complexity and distribution of products generated by IRE-bubble PCR can be exploited and applied to FISH mapping and "chromosome painting" as well as to the generation of STSs targeted to specific chromosomal or subchromosomal regions.

Animals↗

The mouse Fas-ligand gene is mutated in gld mice and is part of a TNF family gene cluster.

The gene for the mouse Fas ligand was cloned and its chromosomal position determined. Fasl was tightly linked to gld (no crossovers in 567 meiotic events) on mouse chromosome 1 and closely linked with a novel member of the same TNF family of ligands, the Ox40 ligand (Ox40l, 1 crossover in 567 meiotic events). Southern blot analysis did not reveal any difference between the Fasl gene from gld and wild-type mice and levels of Fasl mRNA transcripts were similar in PMA and ionomycin induced wild-type and coisogenic gld T cells. Sequence analysis of the gld gene indicated a single amino acid change (Phe Leu) in the COOH terminal portion of this type II transmembrane protein, and COS cells transfected with Fasl cDNA from gld mice failed to induce apoptosis of Fas-expressing target cells. Thus, the data demonstrate that the gld phenotype is the result of a point mutation in the Fasl gene and that Fasl is part of a complex of ligands structurally related to TNF mapping within a small region of mouse chromosome 1.

Amino Acid Sequence↗

Sequential positron emission tomographic evaluations of brain metabolism in acute herpes encephalitis.

This first known positron emission tomography report on metabolic changes in acute herpes simplex virus (HSV-1) encephalitis demonstrates focal hypermetabolism in areas of cerebral cortex adjacent to actively inflamed hippocampus acutely infected with HSV-1. When neuropsychiatric symptoms recurred in a previously healthy 61-year-old patient 1 month after recovering from acute HSV-1 encephalitis, repeat positron emission tomography with fluorodeoxyglucose was helpful in ruling out recurrent active infection by showing marked hypometabolism throughout the previously infected temporal lobe.

Acute Disease↗

Evidence that carbon monoxide is a mediator of critical illness.

We have measured arterial carboxyhemoglobin levels as an indicator of endogenous carbon monoxide production in 32 consecutive admissions to our Surgical Intensive Care Unit. A severity of illness score (APACHE II) was determined for each patient at the same time that blood was drawn. An increase in the APACHE II score indicates a worsening of the patient's condition. There was a positive correlation between arterial carboxyhemoglobin levels and the APACHE II score (p < 0.005). An increase in the white blood cell count indicates an increase in the severity of illness. Carboxyhemoglobin levels correlated with an increase in the white blood cell count to greater than the normal maximum count of 10.5 x 10(3) cells/mm3 within a 24 hour period (p < 0.05). These findings support our hypothesis that endogenously produced carbon monoxide may play a role in the pathogenesis of critical illness.

Carbon Monoxide↗

Objective decision-making following a portal film: the results of a pilot study.

PURPOSE: To discriminate between random and systematic treatment setup errors using portal films. METHODS AND MATERIALS: A bi-dimensional analytic techniques using multiple analyses of variance based on Hotelling's T2 statistics to derive numerical and graphical measures of daily portal film accuracy and precision has been trialed using 88 daily portal films from seven patients' treatment. RESULTS: A demonstration is provided of how a reasonable approximation of random variation from the intended (Simulator) field center, and systematic displacement of the mean position of the portal film centers may be derived from a minimum number of portal films. If a random error as great as 10 mm exists, at least six or seven portal films are considered necessary to reliably detect and quantify the size of a systematic error. CONCLUSION: Our results suggest that a modest systematic error could go undetected until the end of a 5 or 6 week course of treatment if only one portal film is obtained each week. A greater number of portal films should be performed during the first week of treatment to reduce the frequency of such errors. Efforts to separate and quantify both random and systematic errors in setup are worthwhile and will lead to improvements in outcome at the individual patient level and at a departmental level in the development of quality assurance programs.

Humans↗

Molecular basis of myotonic dystrophy: expansion of a trinucleotide (CTG) repeat at the 3' end of a transcript encoding a protein kinase family member.

Using positional cloning strategies, we have identified a CTG triplet repeat that undergoes expansion in myotonic dystrophy patients. This sequence is highly variable in the normal population. PCR analysis of the interval containing this repeat indicates that unaffected individuals have been 5 and 27 copies. Myotonic dystrophy patients who are minimally affected have at least 50 repeats, while more severely affected patients have expansion of the repeat containing segment up to several kilobase pairs. The CTG repeat is transcribed and is located in the 3' untranslated region of an mRNA that is expressed in tissues affected by myotonic dystrophy. This mRNA encodes a polypeptide that is a member of the protein kinase family.

Amino Acid Sequence↗

Retinoic acid stimulates neurite outgrowth in the amphibian spinal cord.

There is increasing evidence that retinoic acid (RA), a vitamin A metabolite, plays a role in the development of the nervous system. Here we specifically test this notion by examining the effect of RA on neurite outgrowth from explanted segments of the axolotl spinal cord. We show that there is a threshold concentration in the region of 0.1-1 nM above which neurite outgrowth is stimulated 4-5 fold. Retinol, by contrast, only stimulated the migration of glial cells from the explants. Using HPLC we demonstrate that RA and retinol are present endogenously in the axolotl spinal cord. In addition, we have identified by immunocytochemistry with antipeptide antibodies the cells of the spinal cord that contain the binding proteins for RA (cellular RA-binding protein; CRABP) and retinol (cellular retinol-binding protein; CRBP). CRABP is found in the axons and CRBP is found in the ependyma and glial cells. These results provide strong evidence for a role for RA in the developing nervous system, and we propose a specific hypothesis involving CRBP, CRABP, retinol, and RA in the control of axon outgrowth in the spinal cord.

Ambystoma↗

Isolation and characterization of irradiation fusion hybrids from mouse chromosome 1 for mapping Rmc-1, a gene encoding a cellular receptor for MCF class murine retroviruses.

An irradiation-reduced somatic cell hybrid mapping panel was constructed of BALB/c mouse Chromosome 1. Nineteen hybrids were selected from a pool of 292 clones to generate a fine structure physical map of the distal 40 cM of the chromosome. The hybrids contain mouse DNA fragments only from Chromosome 1, ranging from approximately 5 cM to approximately 20 cM. Utilizing a viral infectibility assay, a cellular receptor gene, Rmc-1, for the MCF class of murine retroviruses was found to be linked to Lamb2, in the region between the Lamb2 and Bxv-1 loci. In addition, analysis of the hybrid mapping panel resulted in the remapping of three loci, Atpb, Ly-5, and Pmv-24, as compared to the mouse linkage map. Two previously unmapped endogenous proviruses are also putatively assigned positions on the chromosome.

Animals↗

A new robust index for measuring isokinetic trunk flexion and extension. Outcome from a regional study.

The purpose of this study was 1) to provide normative data for isokinetic trunk strength as measured by peak torque, best work repetition, and a new composite variable, "muscle performance index"; and 2) to examine the effects of protocol velocity, subject age, gender, height, and weight on these measurements. A significant decrease in peak torque of extension for females (P less than 0.0001) was seen as protocol velocity increased, but no differences were seen in males. A statistically significant amount (0.5-0.7) of the variance in isokinetic measurements was explained by subject age, height, and weight. Multivariant analysis of covariance showed the effects of these variances to be significant on all measurements except for the muscle performance index, which was unaffected by protocol velocity and remained stable without adjustments for subject age, weight, or height.

Adult↗

Parents' accuracy in estimating child weight status.

Mothers of 107 preschool children estimated their child's weight status, and the accuracy of these estimates was examined. The majority of mothers (72%) were accurate. Of those who were inaccurate, 83% had underestimated the child's weight status, whereas only 17% had overestimated. Mothers of heavier children were more likely to underestimate their child's weight status.

Adult↗

Impaired metabolic function and signaling defects in phagocytic cells in glycogen storage disease type 1b.

Patients with glycogen storage disease (GSD) type 1b (1b), in contrast to patients with GSD type 1a (1a), are susceptible to recurrent bacterial infections suggesting an impairment in their immune system. In this study, phagocytic cell (neutrophil and monocyte) respiratory burst activity, as measured by superoxide anion generation, oxygen consumption, and hexose monophosphate shunt activity, was markedly reduced in both neutrophils and monocytes from GSD 1b patients as compared with either GSD 1a patients or healthy adult control cells. Degranulation, unlike respiratory burst activity, was not significantly different in neutrophils from GSD 1b patients as compared with controls. Both neutrophils and monocytes from GSD 1b patients showed decreased ability to elevate cytosolic calcium in response to the chemotactic peptide f-Met-Leu-Phe. In addition, calcium mobilization in response to ionomycin was also attenuated suggesting decreased calcium stores. Thus, reduced phagocytic cell function in GSD 1b is associated with diminished calcium mobilization and defective calcium stores. Defective calcium signaling is associated with a selective defect in respiratory burst activity but not degranulation.

Calcium↗

Alterations in circulating human chorionic gonadotropin free alpha-subunit in insulin-dependent diabetic pregnancy: correlation with maternal characteristics.

Circulating concentrations of hCG free alpha-subunit (alpha hCG) increase throughout pregnancy. To address the hypothesis that maternal plasma alpha hCG may reflect placental dysfunction and/or adverse perinatal outcome during insulin-dependent diabetic pregnancy, alpha hCG was measured serially throughout gestation, beginning before week 12, with a specific RIA using a monoclonal antibody in 54 insulin-dependent diabetic (randomly assigned to strict and customary glycemic control) and 25 nondiabetic pregnancies. alpha hCG was significantly lower in pregnant insulin-dependent diabetic subjects than in nondiabetics subjects until 24 weeks gestation, after which it was higher until delivery. Plasma alpha hCG stabilized in nondiabetics at 32 weeks, whereas it continued to increase in diabetics until delivery, at which time it was 37% greater than that in nondiabetics (mean +/- SE, 1441 +/- 90 vs. 1052 +/- 78 micrograms/L; P less than 0.002). Values in diabetic subjects assigned to strict control were intermediate between those in diabetic subjects assigned to customary control and nondiabetic subjects. alpha hCG was greater in diabetic subjects with pregestational hypertension or microvascular disease, but not in those with pregnancy-induced hypertension. These findings were independent of the assigned goals of glycemic control. alpha hCG was not correlated with the duration of diabetes or related to premature delivery, fetal distress, birth asphyxia, or macrosomia. Thus, alpha hCG is increased during the third trimester of the type I diabetic pregnancy and is associated with preexisting hypertension and maternal microangiopathy, but is not a predictor of adverse perinatal outcome. Excessive alpha hCG secretion in diabetes may share pathophysiological mechanisms in common with those underlying diabetic microangiopathy.

Adult↗