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Biomedical subjects

J Wilcox

Publications and source records attributed to J Wilcox.

At least 19 recordsLinked to original sources

Synthesis of transforming growth factor-beta 1 by megakaryocytes and its localization to megakaryocyte and platelet alpha-granules.

We have directly demonstrated that megakaryocytes are a major site of synthesis and storage of transforming growth factor-beta 1 (TGF/beta 1) by combined immunohistochemical, immunocytochemical, and in situ hybridization methods. The presence of TGF/beta 1 messenger RNA (mRNA) in mature megakaryocytes in adult rat spleen and bone marrow (BM) was established by in situ hybridization. Localization of TGF/beta 1 protein to intact alpha-granules of megakaryocytes, its putative storage site, was accomplished in glycol-methacrylate embedded porcine BM with an immunoperoxidase technique and light microscopy. The TGF/beta 1 was sequestered in intracytoplasmic granules in a pattern virtually identical to that of another alpha-granule marker protein, fibrinogen. This observation strongly suggests packaging of TGF/beta 1 into this organelle within megakaryocytes. That TGF/beta 1 mRNA was localized to megakaryocytes suggests that the TGF/beta 1 found in the alpha-granules in platelets originates with megakaryocyte synthesis. The alpha-granule localization of TGF/beta 1, as well as fibrinogen, was also demonstrated in isolated platelets at the ultrastructural level by electronmicroscopy (EM) and postembedding colloidal-gold immunocytochemistry, thus directly demonstrating that alpha-granules are the final storage site for TGF/beta 1 in mature platelets.

Animals

The effect of gender and subtype on platelet MAO in alcoholism.

Decreased platelet MAO activity has been identified in male alcoholics with suggestions that this is primarily true of Type 2 alcoholics as defined by criteria from the Stockholm Adoption Study. Little information has been available regarding platelet MAO activity in female alcoholics. This study evaluated a group of 71 alcoholics receiving inpatient treatment, including 16 female alcoholics, for platelet MAO activity compared to controls. Female alcoholic's platelet MAO was significantly lower than controls and not different from activity levels in male alcoholics. Among male alcoholics, both Type 1 and Type 2 subgroups were lower than controls and Type 2 levels did not differ from Type 1 levels. Thus, we were unable to replicate a gender and subgroup low platelet MAO specificity among alcoholics, but did find significant differences between alcoholics and controls.

Adult

Intervening sequence-specific in situ hybridization: detection of the pro-opiomelanocortin gene primary transcript in individual neurons.

We describe here the in situ hybridization procedure which we have used to detect the pro-opiomelanocortin (POMC) gene primary transcript in nuclei of individual neurons in the periarcuate region of the hypothalamus. An exon-intron RNA probe was used to detect POMC primary transcript and mature mRNA in nuclear extracts of nucleic acids using a sensitive S1 nuclease protection assay. The levels per cell of nuclear primary transcript were similar to those seen in the anterior pituitary, suggesting that intervening sequence in situ hybridization should be feasible. A nonrepetitive complementary RNA probe specific for the first intervening sequence of the rat POMC gene (POMC IVS-A) was used to detect the POMC primary transcript in hypothalamic tissue sections by in situ hybridization. The distribution of nuclear localized autoradiographic grains was similar to that previously reported for immunocytochemically defined POMC neurons, suggesting that the procedure is also effective in brain cells.

Arcuate Nucleus of Hypothalamus

Renal excretion and computed tomography enhancement of iotrolan and iopamidol in dogs.

A comparison of the renal excretion and computed tomography (CT) enhancement of nonionic, monomer and dimer contrast media (CM) (iopamidol and iotrolan) was planned to determine if the latter provides additional and prolonged enhancement in dogs. The excretion studies show almost identical performance of both CM with respect to plasma concentration and renal excretion. However, the lower osmolality of iotrolan permits greater renal concentration in the medulla, papilla, and urine. The potential for this further enhancement of the kidney in CT or urography is marginal.

Animals

Diazepam binding inhibitor gene expression: location in brain and peripheral tissues of rat.

Diazepam binding inhibitor (DBI), an endogenous 10-kDa polypeptide was isolated from rat and human brain by monitoring displacement of radioactive diazepam bound to specific recognition sites in brain synaptic and mitochondrial membranes. The cellular location of DBI mRNA was studied in rat brain and selected peripheral tissues by in situ hybridization histochemistry with a 35S-labeled single-stranded complementary RNA probe. DBI mRNA was heterogeneously distributed in rat brain, with particularly high levels in the area postrema, the cerebellar cortex, and ependyma of the third ventricle. Intermediate levels were found in the olfactory bulb, pontine nuclei, inferior colliculi, arcuate nucleus, and pineal gland. Relatively low but significant levels of silver grains were observed overlying many mesencephalic and telencephalic areas that have previously been shown to contain numerous DBI-immunoreactive neurons and a high density of central benzodiazepine receptors. In situ hybridizations also revealed high levels of DBI mRNA in the posterior lobe of the pituitary gland, liver, and germinal center of the white pulp of spleen, all tissues that are rich in peripheral benzodiazepine binding sites. The tissue-specific pattern of DBI gene expression described here could be exploited to further understand the physiological function of DBI in the brain and periphery.

Animals

Examination of the nephrographic potential of iotrol by computed tomography.

The x-ray attenuation of the renal cortex of dogs, as determined by computed tomographic (CT) scanning, was measured over a three-day period after an intravenous bolus of 600 mg I/kg of iotrol or iopamidol. A slightly higher density observed 24 hours after injection of iotrol was not considered significant, and was not considered sufficient to warrant clinical application of iotrol for specific, prolonged renal enhancement.

Animals

Metabolism and DNA-binding in vivo of aflatoxin B1 in medaka (Oryzias latipes).

1. The medaka (Oryzias latipes), a small aquarium fish, was shown to possess the capacity to rapidly activate AFB1 in vivo at 25 degrees C to intermediates that bind to DNA. 2. The dose-response for in vivo AFB1-DNA binding was linear over the range 70-550 micrograms AFB1/kg body weight. 3. Maximum binding occurred within the first 24 hr after i.p. injection of [3H]AFB1, followed by a rapid loss of adducts. 4. Aflatoxicol (AFL) and unreacted AFB1 were found by HPLC analysis to be the major products excreted into water after AFB1 exposure, with excretion of AFL as early as 2 min after AFB1 injection. 5. These studies show that medaka possess enzymatic systems similar to rainbow trout (Salmo gairdneri) for biotransformation of AFB1 to the epoxide and to other phase I and phase II metabolites.

Aflatoxin B1

Rate of clearance of intrathecal iopamidol in the dog.

The incidence of post-myelographic side-effects has been significantly reduced since the advent of the new generation of water-soluble, non-ionic, contrast media (CM). One of these CM, iopamidol, has recently been released for clinical intrathecal use in Australia. Clinical evaluations have shown iopamidol to give excellent diagnostic results. However, many adverse side-effects have been cited in the literature. Reactions to iopamidol following myelography are thought to be due to the lack of a physiologic barrier between the extracellular fluid of the brain parenchyma and the CSF, thereby allowing CM to penetrate the brain parenchyma tissue, following subarachnoid injection. This study investigates the rate of clearance of intrathecal iopamidol from the brain in dogs by performing coronal CT scans at intervals over a 48 h, post-injection period. Analysis of similar regions of interest (ROI) for each time period indicate that iopamidol can be detected in canine brains for at least 48 h following intrathecal injection (P less than 0.05). Furthermore, the disappearance of iopamidol from the brain parenchyma is approximately logarithmic in form, with a half-life of approximately 22 h.

Animals

Effects of intracarotid ionic and non-ionic contrast material on the blood-brain barrier in a rabbit model.

A rabbit model was used to assess the effects of intracarotid injections of ionic monomer (meglumine iothalamate), non-ionic monomer (iohexol, iopromide), and non-ionic dimer (iotrol) contrast materials on the blood-brain barrier. The degree of blood-brain barrier damage was assessed qualitatively using Evans' blue dye, and quantitatively by calculating the difference in pertechnetate uptake between injected and non-injected hemispheres. The results showed that the non-ionic dimer, iotrol, had the least effect on the blood-brain barrier, and that although iopromide and iohexol produced greater damage than iotrol, the ionic compound, meglumine iothalamate, caused the greatest disruption to the blood-brain barrier. The implications of these findings are discussed.

Animals

Effects of intracarotid ioxaglate on the normal blood-brain barrier. A comparison of two animal models.

Using two different models, the effect on the blood-brain barrier of intracarotid injections of sodium/meglumine ioxaglate at similar iodine concentrations (280 mgI/ml) was investigated. In both models the degree of blood-brain barrier damage was assessed visually using Evans' Blue stain. Quantitative assessment of blood-brain barrier disruption was made by contrast enhancement as measured by CT of the dog brain, and by 99mTc-pertechnetate uptake by the brain in the rabbit model. No Evans' Blue staining was observed in any study using the canine/CT model. Slight staining was observed in two studies with ioxaglate using the rabbit/pertechnetate model. Statistical analysis of results from the canine/CT model did not detect any damage to the blood-brain barrier with either ioxaglate or saline control studies (P greater than 0.1). However, in the rabbit/pertechnetate model a slight increase in disruption of the blood-brain barrier was observed with ioxaglate compared with control studies, but this was only significant at the 0.1 level. The results suggest that the rabbit/pertechnetate model is a more sensitive measure of blood-brain barrier disruption than the canine/CT model. This study also demonstrates that blood-brain barrier disruption following intracarotid injection of ioxaglate is minimal.

Animals

Metabolism of an ingested serine load in psychotic and nonpsychotic subjects.

Our previous studies have shown that in psychotics, the plasma serine level is abnormally high and that plasma serine hydroxymethyltransferase (which cleaves serine to glycine) activity is abnormally low as compared with that in nonpsychotic subjects. In this study, psychotic and nonpsychotic subjects ingested a large bolus of L-serine (4 mM/kg) at breakfast and blood was drawn before breakfast, 2 hr, 4 hr, and 6 hr after serine ingestion. Baseline serine and SHMT activity differentiated between psychotics and nonpsychotics with high degrees of significance (p less than 0.0001) and p less than 0.01, respectively). Plasma serine levels 2 hr after serine ingestion were significantly higher (p less than 0.01) in nonpsychotics as compared with psychotics. Elimination of serine in psychotics was bimodal and was significantly different from that of nonpsychotics (p less than 0.0079, Moses test). These findings provide additional evidence for abnormal serine metabolism in psychotic patients.

Age Factors

Differentiation of psychotic from nonpsychotic depression by a biological marker.

In a study of fasting plasma serine levels (PSL) previously shown to be a biological marker for psychosis, we found significantly higher (P = 0.0008) PSL in 18 psychotic depressives when compared to 22 nonpsychotic depressives. Similarly the activity of the enzyme serine hydroxymethyltransferase (SHMT) which cleaves serine to glycine, was significantly lower (P less than 0.0001) in psychotics than in nonpsychotics. The difference between psychotic and nonpsychotic depressives were not attributable to age, sex or drug intake. This finding is in support of the hypothesis that these two types of depressions are qualitatively distinct from each other.

Adult

Psychopathology and narcolepsy.

Twenty-eight patients with narcolepsy are compared to a sex- and age-matched control population with regard to the frequency of psychiatric symptoms. The occurrence of psychiatric syndromes in narcolepsy patients is also examined. The frequency of psychiatric illness in patients with cataplexy or hypnagogic/hypnopompic hallucinations is evaluated as well. Analysis with the Mann-Whitney test found a significant association between narcolepsy and the symptoms of schizophrenia. Premorbid history of organic brain disease was associated with mental illness, but was nonspecific for psychopathology.

Adolescent