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Biomedical subjects

E Murray

Publications and source records attributed to E Murray.

At least 91 records · Page 5Linked to original sources

Rapid, simple identification of individual osteoblastic cells and their specific products by cell blotting assay.

Biochemical and molecular biological studies of osteoblastic cell function and hormonal regulation are frequently confounded by the inherent cellular heterogeneity and phenotypic instability of existing in vitro and in vivo model systems. A new technique (derived from Western blotting or antibody-based detection of protein molecules bound to nitrocellulose paper) is described for identification of individual cells which synthesize osteoblast-specific gene products (bone Gla-protein, type I collagen, and alkaline phosphatase) or produce cAMP in response to parathyroid hormone (PTH) or isoproterenol. Dispersed primary neonatal rat calvariae or osteogenic sarcoma cells were "plated" on Immobilon-P (a hydrophobic transfer membrane with very high protein-binding capacity) for 30 minutes to several hours, followed by agonist treatment, formalin fixation, hematoxylin staining, and immunostaining with a battery of antibodies specific for osteoblastic products. Individual cells and their secretory zones were visualized by light microscopy and counted. Treatment with PTH with or without isoproterenol resulted in increases in the percentages of osteoblastic cells elaborating cAMP, as well as the intensity of immunostaining, but had no effects on MCF-7 cells, a nonosteoblastic breast carcinoma control line. The percentage of cells within each primary osteoblastic cell population isolated or rat osteogenic sarcoma cell clone (G2 or C12) that elaborated bone-specific proteins or that generated cAMP in response to PTH varied with time and the individual cellular preparation, reconfirming the cellular heterogeneity of these systems. This method, in conjunction with techniques such as in vitro hybridization, should prove useful in characterizing discrete osteoblastic bone cell subpopulations and in clarifying mechanisms of hormonal regulation by local and systemic agents.

Alkaline Phosphatase↗

Structural family versus psychodynamic child therapy for problematic Hispanic boys.

Structural family therapy, psychodynamic child therapy, and a recreational control condition were compared for 69 six-to-twelve-year-old Hispanic boys who presented with behavioral and emotional problems. The results suggest that the control condition was significantly less effective in retaining cases than the two treatment conditions, which were apparently equivalent in reducing behavioral and emotional problems as well as in improving psychodynamic ratings of child functioning. Structural family therapy was more effective than psychodynamic child therapy in protecting the integrity of the family at 1-year follow-up. Finally, the results did not support basic assumptions of structural family systems therapy regarding the mechanisms mediating symptom reduction.

Child↗

A long-term metabolic study to assess the nutritional value of and immunological tolerance to two soy-protein concentrates in adult humans.

Seventeen healthy young adult men participated in a long-term metabolic study (11 wk) to evaluate the tolerance to and protein nutritional value of two commercially produced soy-protein concentrates. Danpro-S (nine subjects) and Danprotex-H 40 (eight subjects) (Aarhus Oliefabrik A/S, Aarhus, Denmark). Each test protein (0.8 g.kg-1.d-1) served as the sole source of dietary nitrogen. Subjects remained healthy, no problems of clinical or metabolic significance appeared, body weight remained constant, and body composition indices (lean body mass by H2(18)O dilution and creatinine excretion) and basal metabolic rate did not change. N balances fluctuated around body N equilibrium. Immunological studies confirmed the absence of any allergic responses in these subjects. It is concluded that these protein concentrates can be consumed as the sole source of dietary protein for protein nutritional maintenance and with excellent tolerance.

Adult↗

Effect of 6-phosphogluconate on phosphoglucose isomerase in rat brain in vitro and in vivo.

The activity of phosphoglucose isomerase, its kinetic properties, and the effect of 6-phosphogluconate on its activity in the forward (glucose 6-phosphate----fructose 6-phosphate) and the reverse (fructose 6-phosphate----glucose 6-phosphate) reactions were determined in adult rat brain in vitro. The activity of phosphoglucose isomerase (in nmol/min/mg of whole brain protein) was 1,865 +/- 20 in the forward reaction and 1,756 +/- 32 in the reverse reaction at pH 7.5. It was 1,992 +/- 28 and 2,620 +/- 46, respectively, at pH 8.5. The apparent Km and Vmax of phosphoglucose isomerase were 0.593 +/- 0.031 mM and 2,291 +/- 61 nmol/min/mg of protein, respectively, for glucose 6-phosphate and 0.095 +/- 0.013 mM and 2,035 +/- 98 nmol/min/mg of protein, respectively, for fructose 6-phosphate. The activity of phosphoglucose isomerase was inhibited intensely and competitively by 6-phosphogluconate, with an apparent Ki of 0.048 +/- 0.005 mM for glucose 6-phosphate and 0.042 +/- 0.004 mM for fructose 6-phosphate as the substrate. With glucose 6-phosphate as the substrate, at concentrations from 0.05 to 0.5 mM, the activity of the enzyme was inhibited completely in the presence of 0.5-2.0 mM 6-phosphogluconate. With 0.05-0.2 mM fructose 6-phosphate as the substrate, it was inhibited greater than or equal to 85% at the same concentrations of the inhibitor. No significant changes were observed in the values of Km, Vmax, and Ki for phosphoglucose isomerase in the brain of 6-aminonicotinamide-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS)

6-Aminonicotinamide↗

The relationship between visual-perceptual motor abilities and clumsiness in children with and without learning disabilities.

One visual-perceptual test, four visual-motor tests, and a test of motor impairment were administered to 22 children with learning disabilities and 22 children without learning disabilities, aged 5 to 8 years. The children with learning disabilities were divided into two groups--"clumsy" and "nonclumsy"--based on their scores on the motor impairment test. It was hypothesized that the clumsy children with learning disabilities would score significantly lower on visual-perceptual and visual-motor tests than the nonclumsy children with learning disabilities who, in turn, would score significantly lower than the children without learning disabilities. It was further hypothesized that there would be a significant correlation between the degree of clumsiness and the degree of visual-perceptual and visual-motor deficit. Analysis of the data indicated that, as expected, the clumsy children with learning disabilities scored significantly lower than the children without learning disabilities (the control group). There was no significant difference between the clumsy and nonclumsy children with learning disabilities or between the nonclumsy children with learning disabilities and the control group. Degree of clumsiness significantly correlated with scores on four of five tests. Results are discussed in terms of subtypes of learning disabilities and sample size.

Analysis of Variance↗

[Lactose tolerance and milk consumption: myths and realities].

The disaccharide lactose, the principal carbohydrate of animal milks, requires the enzyme lactase to split it to glucose and galactose. Undigested lactose passes to the colon where fermentation produces hydrogen and short-chain fatty acids that can cause abdominal distention, pain and sometimes diarrhea. Persistence of intestinal lactase after early childhood, is inherited as a highly-penetrant autosomal dominant genetic characteristic. On the basis of a review of over 560 references, all available data on the primary loss of intestinal lactase in Latin American populations are presented in tabular form. Prevalence of lactose non-digesters in Latin American populations ranges from 45% to 100%. However, this is not a reliable predictor of the acceptability of milk and milk products containing lactose. Milk is being used successfully for the supplementary feeding of children worldwide, and most lactose non-digesters can tolerate at least 240 ml of milk or the lactose equivalent in other products. Lactose maldigestion does not interfere with the absorption of the protein and essential micronutrients in milk. Information is provided on the lactose content of milk and milk products, on the usual milk consumption of Latin American populations, and on worldwide experimental and field observations of milk acceptability. Both adaptation to continued use of milk and milk products and relationships of milk use to various disease states in which intestinal lactase activity may be reduced are discussed. Some types of yoghurts are better tolerated because of the lactase activity of the bacteria used in their fermentation. For unusually intolerant individuals commercial enzyme preparations are available for addition to milk products but for most persons the additional cost is unnecessary.

Adult↗

Characterization of a human osteoblastic osteosarcoma cell line (SAOS-2) with high bone alkaline phosphatase activity.

We characterized the alkaline phosphatase activity of the human osteogenic sarcoma cell line, SAOS-2, and studied the regulation of this enzyme and 3',5'-cyclic adenosine monophosphate levels by 1,25-dihydroxyvitamin D3 and triamcinolone acetonide. We report that the basal alkaline phosphatase activity of SAOS-2 cells was 100-1000 times greater than that of other established human osteogenic sarcoma cell lines. The enzymatic activity was thermolabile, could be inhibited by levamisole and L-homoarginine, but not by L-phenylalanine, and was immunoprecipitable with anti-bone/liver/kidney, but not with anti-placental antibody, confirming that it is the tissue-unspecific or bone/liver/kidney isoenzyme. However, in contrast to other established human osteosarcoma cell lines (TE-85, SAOS-1), in which alkaline phosphatase activity is stimulated several-fold by the steroid hormones 1,25-dihydroxyvitamin D3 and hydrocortisone, the alkaline phosphatase activity of SAOS-2 cells was not affected by 1,25-dihydroxyvitamin D3 treatment despite the presence of classical receptors for this hormone. Furthermore, administration of the potent glucocorticoid analogue, triamcinolone acetonide, induced only a modest increase in activity. The SAOS-2 cell line expressed low basal cAMP levels (28 pmol/10(6) cells) which could be increased 25-40 times by pretreatment with parathyroid hormone. However, unlike other osteoblastic models, in which PTH-induced cAMP stimulation is modulated by 1,25-dihydroxyvitamin D3 and glucocorticoids, neither of these hormones had an effect on the PTH-stimulated cAMP levels in SAOS-2 cells. We conclude that the SAOS-2 cell line is an osteoblastic cell model which expresses high levels of tissue-unspecific alkaline phosphatase activity and exhibits limited responsiveness to two steroid hormones.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaline Phosphatase↗

Anesthetic induction with fentanyl and intravenous lidocaine for coronary artery surgery.

In a randomized, double-blind trial, 59 patients undergoing coronary artery surgery received fentanyl 10, 15, or 25 microg/kg infused over 5 minutes for anesthetic induction. Half of the patients received intravenous lidocaine, 1.5 mg/kg, 1 minute before laryngoscopy. Efficacy of induction as judged by loss of consciousness was evaluated, and hemodynamic values during induction, laryngoscopy, and tracheal intubation were recorded each minute for 10 minutes. Plasma fentanyl concentrations were determined after termination of the fentanyl infusion. Opioid induction with fentanyl was successful in 90% (18 of 20) of patients receiving 25 microg/kg, 89% (17 of 19) of patients receiving 15 microg/kg, but only 55% (11 of 20) of patients receiving 10 microg/kg (P < .01). While plasma fentanyl concentrations were proportional to the dose infused (25 ng/mL, 18 ng/mL, and 14 ng/mL in the 25, 15. and 10 microg/kg fentanyl groups, respectively), there was no relationship between plasma fentanyl concentration and hemodynamic response to laryngoscopy or intubation. Opioid induction caused a gradual decrease in blood pressure that was restored with intubation. Lidocaine partially blocked this restoration (systolic blood pressure 122 +/- 5 v 138 +/- 5 mmHg, lidocaine v placebo, 1 minute after laryngoscopy, P < .05). Fentanyl, 15 or 25 microg/kg, intravenously, is an effective induction agent for patients with coronary artery disease. Supplementation with intravenous lidocaine, 1.5 mg/kg, will obtund the increase in blood pressure that occurs with laryngoscopy and intubation and help prevent infrequent hypertensive responses seen with this opioid technique.

Adjuvants, Anesthesia↗

T6+ and HLA-DR+ cell numbers in epidermis of immunosuppressed renal transplant recipients.

The increased susceptibility of the skin of chronically immunosuppressed individuals to viral infections and sunlight-induced malignancies suggests specific drug-induced, dysfunction of local immune mechanisms within the sun-exposed skin of these individuals. To help understand the effect of immunosuppressive therapy alone in the absence of ultraviolet light on the immune system of skin, biopsies were collected from non-sun-exposed buttock skin of control, healthy volunteers and kidney transplant recipients immunosuppressed with either azathioprine/prednisone or cyclosporin A/prednisone and examined for incidences of T6+, and HLA-DR+ cells. No significant differences in the incidences of these 2 cell types were found (a) between control individuals and transplants recipients, (b) between transplant recipients receiving either of the immunosuppressive drug regimes, or (c) between transplant recipients who either had or had not developed skin cancer.

Adolescent↗

Neurotropic Melanoma. A variant of desmoplastic melanoma.

In ten cases of neurotropic melanoma, patients presented with nodules composed of amelanotic, deeply infiltrating neuroid fascicles. Only four cases were clinically suggestive of melanoma. In eight of the tumors, a precursor lesion was found histologically and provided a major clue to the diagnosis. In seven cases, the dysplastic precursor was lentiginous, while a superficial spreading pattern was present in one. Initial surgery was often inadequate because of the difficulty in defining tumor margins and the lack of pigment. In seven of the tumors, S100 protein was demonstrated within the invasive spindle cell component by the immunoperoxidase method. This finding was negative in three cases, two of which showed positive staining of the precursor and nerve filaments, indicating that the absence of S100 protein cannot be used as an exclusion criterion for neurotropic melanoma.

Adult↗

Effects of exposure to ultraviolet light on epidermal Langerhans cells and melanocytes in Australians of aboriginal, Asian and Celtic descent.

The effects of 3 daily, 7-min exposures to artificial ultraviolet light (UVL), designed to simulate natural sunlight, on epidermal Langerhans cells (LC) and melanocytes were studied in 17 healthy Australian volunteers of differing skin pigmentation. Six were of Celtic, 6 of Asian and 5 of Aboriginal descent. LC were visualized using the immunofluorescence method for HLA-DR and T6 antigens, and the histochemical method for ATPase. UVL induced a transient reduction in the LC population density and an increase in the number of melanocytes in all subject groups. The reduction in number of immunocompetent LC or the disruption of their surface markers was greatest in the Celtic subjects, who had the fairest skin, and least in the Aboriginal and Asian subjects, who had the darkest skin. However, neither the inherently dark skin pigmentation nor the UVL-induced increase in pigmentation were sufficient to prevent the depletion of immunocompetent epidermal LC. Non-dendritic, rounded cells and very large dendritic, non-LC, which were present in the epidermis of some subjects, were stimulated to increase in number by exposure to UVL. The identity and function of these cells is uncertain and they require further investigation.

Adult↗

Comparison of cefixime and co-trimoxazole in acute uncomplicated urinary tract infection. A double-blind general practice study.

Five hundred and twenty-eight patients with presumptive acute uncomplicated urinary tract infection (UTI) were randomly assigned to receive cefixime 400 mg once daily, cefixime 200 mg twice daily or co-trimoxazole 2 tablets twice a day for 10 days; 477 completed at least 5 days of therapy. Of the patients 342 (65%) had positive baseline urine cultures, yielding 353 pathogens. A microbiological response was determined for 280 pathogens (79%), eradication being observed in over 94% of isolates; 153 pathogens (43%) were sensitive to both cefixime and co-trimoxazole and eradication was observed in over 96% of cases. Clinical response correlated well with microbiological response. The incidence of diarrhoea and stool changes was higher (P less than 0.005) in the patients who received cefixime once daily than in the other groups. There was a significantly higher incidence of stool changes with cefixime twice daily than with co-trimoxazole (P less than 0.05), but these did not necessitate discontinuation of therapy. Nausea was commoner with co-trimoxazole (P less than 0.05). The majority of pathogens isolated were Escherichia coli, Proteus mirabilis and staphylococci. Approximately 24% of E. coli were resistant in vitro to co-trimoxazole (P less than 0.005). Cefixime 200 mg twice daily is an effective and safe alternative to co-trimoxazole in the management of acute uncomplicated UTI.

Adult↗

Superior oblique paralysis. A review of 270 cases.

In 270 patients with superior oblique paralyses treated between 1973 and 1984, congenital and traumatic causes were most frequent, and one fourth of all traumatic cases had bilateral involvement. Among the diagnostic features distinguishing bilateral from unilateral paralysis were a right hypertropia in left gaze and left hypertropia in right gaze, and a positive Bielschowsky test on tilting the head toward either shoulder. However, absence of either sign did not exclude bilateral paralysis. Large excyclotropia and a V-pattern esotropia are suggestive of but not diagnostic for bilateral paralysis. Complaints about cyclotropia are limited to acquired paralysis. Cyclotropia in the normal eye, head tilt toward the involved side, or absence of any abnormal head posture limits the diagnostic value of these associated signs. Overshoot of the contralateral superior oblique occurred in 19% of the patients and is thought to be caused by contracture of the ipsilateral superior rectus muscle. Surgical treatment in 112 patients resulted in an 85% cure rate with an average of 1.45 operations per patient.

Adolescent↗

Demonstration of insulin receptors and modulation of alkaline phosphatase activity by insulin in rat osteoblastic cells.

Osteoporosis is a known complication of diabetes mellitus, suggesting a role for insulin in bone homeostasis. We studied insulin receptors and insulin action in the osteoblast-like rat osteogenic sarcoma cell line ROS 17/2.8. These cells share many common features with the osteoblast, such as 1,25-dihydroxyvitamin D3 receptors, PTH receptors, and 1,25-dihydroxyvitamin D3-induced modulation of alkaline phosphatase activity and osteocalcin. Competition binding studies revealed high affinity insulin receptors, with an ED50 for insulin of 1 nM. The receptors were highly specific for insulin, with 60% inhibition of insulin binding by an antireceptor antibody, no competition by epidermal growth factor, and an ED50 of 300 nM for proinsulin. Steady state maximal insulin binding was obtained by 40 min at 37 C, and insulin degradation, as measured by trichloroacetic acid solubility, was 1%/h at 37 C. ROS cells readily internalized insulin, and under steady state binding conditions at 37 C, 56% of the cell-associated radioactivity consisted of intracellular material. Chloroquine (100 microM) inhibited intracellular processing of insulin, leading to a 300% increase in cell-associated insulin by 2 h (37 C). Photoaffinity labeling of the insulin receptor with the photosensitive analog of insulin, B2 (2-nitro-4-azidophenyl-acetyl)des-pheB1-insulin, followed by solubilization and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, revealed specific bands of 125K and 430K mol wt under reducing and nonreducing conditions, respectively. Thus, the structure of insulin receptors in ROS cells appears comparable to that of insulin receptors of known target tissues. Insulin action was also examined. Insulin did not stimulate [2-3H]deoxyglucose uptake or [1-14C]leucine incorporation into protein. In contrast, physiological concentrations of insulin inhibited alkaline phosphatase activity in nonconfluent cells. After exposure to insulin for 24 h, alkaline phosphatase activity was decreased compared to basal by 39.5% and 50% with 5 and 50 ng/ml insulin, respectively. In conclusion, ROS cells bind insulin to specific receptors that are similar to insulin receptors on other target tissues; receptors internalize insulin, which is then processed through a chloroquine-sensitive pathway; insulin does not affect membrane substrate transport; and insulin does inhibit the activity of an enzyme that is important in bone metabolism. ROS cells represent a model for studying insulin effects on bone.

Affinity Labels↗

Current research on immunopathology of melanoma: analysis of lymphocyte populations in relation to antigen expression and histological features of melanoma.

Immunoperoxidase localization of monoclonal antibodies in sections of melanoma has been used to identify histological features which may be of prognostic importance in melanoma, in particular whether certain structures on melanoma cells may determine the degree and nature of lymphoid infiltrates and whether these may be related to prognosis. Monoclonal antibodies to lymphocyte subpopulations were used to identify and quantitate lymphoid infiltrates in 15 primary melanomas and 8 cutaneous metastases. These were correlated with histological features identified in routine sections. There was a wide variation in the numbers of mononuclear cells associated with both primary and metastatic melanoma. T cells and to a lesser extent macrophages accounted for the majority of the cells, B cells and natural killer (NK) cells for only 2% of the infiltrates. The Leu 3a (helper) T cell subpopulation predominated in primary tumours, OKT8 positive (suppressor/cytotoxic) cells in metastases. Infiltrates in which OKT8 cells predominated were associated with ulceration, a high mitotic rate and thick primary tumours. The converse applied to Leu 3a infiltrates. Infiltrates with a high proportion of Leu 3a positive cells tended to be associated with Thy-1 positive, DR antigen negative tumours. Thy-1 antigens were predominantly expressed on primary tumours and rarely on metastases whereas the converse applied to expression of the acute lymphoblastic leukemia antigen (CALLA) on melanoma cells. These findings suggest that certain melanoma antigens may be related to the nature of the lymphoid infiltrate associated with melanoma and possibly with the behaviour of the tumour in the host. They further suggest that identification of cell surface structures and lymphoid infiltrates by these techniques may be a valuable extension of routine histopathological assessment of prognosis in melanoma.

Antibodies, Monoclonal↗

Differential spread of blockade of touch, cold, and pinprick during spinal anesthesia.

The differential levels of sensory blockade of pinprick, cold, and touch were monitored throughout the course of spinal anesthesia administered to 50 patients to determine variations in the degree of spread during onset, plateau, and regression, and to establish the effects of epinephrine and the effect of posture during injection. A significant difference was observed between the dermatomal level of sensory loss of touch and the dermatomal level of loss of either pinprick or cold during onset, at 5 min in patients given tetracaine with epinephrine, at time of maximum spread in patients given tetracaine with epinephrine or in the sitting position, and in all groups during regression. Loss of touch began later, never extended as far cephalad, and regressed sooner. The extent of this difference was greatest during regression, when the anesthetic was given to patients in the sitting position, after epinephrine. The level at which the sense of touch was lost seemed to mark the limits of the zone of solid spinal anesthesia; these limits could not be assessed effectively using pinprick. We propose that loss of touch sensation be used to assess whether anesthesia is adequate to avoid tourniquet pain. If there is loss of touch sensation above the L1 dermatome, it is unlikely that tourniquet pain will occur.

Adult↗

A long-term metabolic balance study in young men to assess the nutritional quality of an isolated soy protein and beef proteins.

To evaluate the capacity of an isolated soy protein to maintain long-term protein nutritional status in healthy young adult men, an 84-day metabolic balance experiment was conducted in eight subjects. The sole source of protein intake was provided by the isolated soy protein, given at a level of 0.8 g (N X 6.25) per kg per day. In a second and similar study, four young men received 0.8 g protein and three subjects 0.68 g protein per kg per day from beef proteins for 60 to 81 days. Body weight, nitrogen balance, blood chemistries, and body composition (whole body 40K) were monitored throughout each study. Body nitrogen balances were maintained within the range of N equilibrium in both diet groups. Body cell mass, as judged from 40K measurements, did not reveal any deterioration in protein nutritional status. These observations confirm the prediction, derived from previous short-term. N balance studies, that the nutritional quality of isolated soy protein is high and that this plant protein can serve as the sole source of essential amino acids and nitrogen for protein maintenance in adults.

Adolescent↗