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Biomedical subjects

M A Miller

Publications and source records attributed to M A Miller.

At least 271 records · Page 15Linked to original sources
PubMed1994

Laminin mediates the restitution of rat gastric mucosa in vitro.

Restitution, the rapid re-establishment of mucosal integrity following damage, involves cell migration and can be monitored by measuring transmucosal potential difference of tissue mounted in an Ussing chamber. The involvement of extracellular matrix proteins and matrix receptors was examined in the restitution of rat gastric mucosa. Undamaged mucosa maintained a potential difference of -32.7 +/- 2.2 mV for several hours. Mucosal exposure to 0.6 M NaCl for 1 min reduced this to -3.3 +/- 1.4 mV in 2-3 min. Thereafter, the potential difference returned in 60 min to plateau at -28.9 +/- 1.3 mV (88.5 +/- 3.6% of pre-exposure). Tissues mucosally treated with 1:100 anti-laminin antiserum maximally recovered following damage to 65.6 +/- 6.6% of pre-exposure potential difference (PD), while those treated with 1:100 anti-collagen IV or anti-fibronectin antisera recovered to 88.8 +/- 9.7% and 86.3 +/- 3.2%, respectively. Only the anti-laminin result was significantly different from controls. The anti-laminin effect was abolished by pre-incubation of the anti-laminin antiserum with purified rat laminin, suggesting that the effect was laminin specific. In experiments involving matrix protein receptors, tissues treated with alpha-lactalbumin, a protein altering the substrate specificity of cell surface laminin receptor/enzyme beta-1,4-galactosyltransferase, maximally recovered following damage to only 49.3 +/- 7.7% of pre-exposure PD, which was significantly different from controls, while those treated with anti-beta 1 integrin recovered to 85.0 +/- 9.7%. Our data suggest that laminin is involved in mediation of gastric mucosal restitution, possibly via beta-1,4-galactosyltransferase.

Animals↗
PubMed1994

Angiotensin II suppression is a major factor permitting excretion of an acute sodium load in humans.

We examined the role of circulating angiotensin II (ANG II) in the excretion of an acute Na+ load in eight healthy subjects given 2 liters 0.9% saline in a placebo-controlled crossover study. On the control day, plasma ANG II decreased to 40-60% of basal values and 35 +/- 9 (SE) mmol of Na+ was excreted in the 5 h after the start of saline infusion. When ANG II was infused to maintain plasma ANG II levels at around basal values (6.6 +/- 1.6 pmol/l), only 7 +/- 8 mmol of Na+ was excreted in the same period (P < 0.05). In a previous similar study in which the fall in aldosterone was prevented by infusion of aldosterone, 16 +/- 16 mmol of Na+ was excreted vs. 36 +/- 16 mmol on the control day in comparable 5-h periods. Suppression of ANG II is one of the major factors permitting the acute increase in Na+ excretion after an intravenous Na+ load. ANG II has direct actions on Na+ excretion in addition to its effects on aldosterone.

Adult↗
PubMed1994

Blood pressure and endocrine responses to changes in dietary sodium intake in cardiac transplant recipients. Implications for the control of sodium balance.

BACKGROUND: The role of cardiac extrinsic innervation in the regulation of sodium balance and blood pressure is controversial. METHODS AND RESULTS: We performed a double-blind study of endocrine and blood pressure responses to 5 days of low- (LS, 10 mmol/d) and 5 days of high- (350 mmol/d) sodium intake in 12 cardiac transplant recipients, 12 matched healthy subjects, and 12 matched subjects with untreated essential hypertension. In transplant recipients on low sodium, supine blood pressure was 137/94 +/- 8/4 (mean +/- SEM) mm Hg and plasma atrial natriuretic peptide (ANP) was 59.3 +/- 6.3 pg/mL; on high sodium, blood pressure was 148/97 +/- 5/3 mmHg (P < .05 for systolic pressure versus LS), and ANP was 94.3 +/- 10.6 pg/mL (P < .01 versus LS), respectively. Plasma ANP for those on each diet was significantly higher in the cardiac transplant recipients than in healthy or hypertensive controls; relative changes in plasma ANP in changing from low- to high-sodium diet were similar in each group. Urinary sodium excretion by the fifth day of each diet was similar in each group. Suppression of plasma renin activity and aldosterone by high-sodium diet was blunted in cardiac transplant recipients compared with healthy subjects (respectively, plasma renin activity: 1.41 +/- 0.30 versus 0.68 +/- 0.21 ng.mL-1 x h-1, P < .05; aldosterone: 391 +/- 35 versus 166 +/- 21 pmol/L, P < .05). CONCLUSIONS: These results suggest that extensive denervation of the heart does not result in major abnormalities in regulation of large changes in sodium intake and that intact cardiac innervation is not required for plasma ANP responses to altered sodium intake. Blood pressure after cardiac transplantation is sensitive to reduced sodium intake.

Atrial Natriuretic Factor↗
PubMed1994

Immunohistochemical staining of feline malignant fibrous histiocytomas.

Malignant fibrous histiocytoma was diagnosed in seven cats from biopsy specimens received at the University of Missouri Veterinary Medical Diagnostic Laboratory during a 4-year period from 1987-1991. Tissue blocks from formalin-fixed specimens were resectioned and stained for type I (AE1) and type II (AE3) cytokeratins, desmin, S100 protein, vimentin, and alpha 1-antitrypsin by the avidin biotin peroxidase complex method with diaminobenzidine (DAB) chromogen. None of the tumors stained positively for alpha 1-antitrypsin. Four of seven of the tumors had similar immunohistochemical staining results, with positive staining for type I and type II cytokeratins, desmin, S100 protein, and vimentin. Of the remaining three, one stained positively only for S100 protein and vimentin; one stained positively for vimentin only; and one was negative for all six antigens. Based only on immunohistochemical staining results, three of the tumors could possibly be reclassified: one as a melanoma, one as a probable fibrosarcoma, and one as unknown. These results also indicate that feline malignant fibrous histiocytomas show a diversity of intermediate filament expression, as do human tumors. Our results also do not support the theory that malignant fibrous histiocytomas are of histiocytic origin.

Animals↗
PubMed1994

Galanin in the bed nucleus of the stria terminalis and medial amygdala of the rat: lack of sexual dimorphism despite regulation of gene expression across puberty.

Neurons in the bed nucleus of the stria terminalis (BNST) and the medial amygdala (AMe) coexpress vasopressin and galanin (GAL) in the adult male rat. Here, we have asked whether GAL gene expression, like vasopressin gene expression in these same neurons, exhibits sexual dimorphism and whether GAL pathways in the BNST and AMe are activated with puberty in female rats as we have previously observed in male rats. In Exp 1, in situ hybridization histochemistry and quantitative autoradiography were used to compare GAL gene expression in the BNST and AMe of prepubertal (24-day-old) and adult (90-day-old) male and female rats. In the BNST, both the number of GAL mRNA-expressing neurons (F = 41.98; P < or = 0.0001; males, P < or = 0.007; females, P < or = 0.001) and the intensity of labeling (F = 40.35; P < or = 0.0001; males, P < or = 0.004; females, P < or = 0.002) were significantly increased in adult compared to prepubertal animals of both sexes. In the AMe of both males (P < or = 0.001) and females (P < or = 0.001), the intensity of labeling was significantly enhanced across puberty (F = 66.29; P < or = 0.0001); however, the number of GAL mRNA-expressing neurons in this region did not change. We found no evidence for sexual dimorphism of GAL gene expression in either brain region. In Exp 2, we replicated our observations of a lack of sexual dimorphism of GAL gene expression in the BNST of adult male and female rats. These findings are consistent with the hypothesis that GAL neurons in the BNST and AMe are steroid sensitive in both sexes. However, our failure to detect any differences in either the number of GAL mRNA-expressing neurons or the level of expression between male and female rats at either age indicates that these pathways do not exhibit sexual dimorphism.

Amygdala↗
PubMed1994

Estrogen receptor and neurotensin/neuromedin-N gene expression in the preoptic area are unaltered with age in Fischer 344 female rats.

The sensitivity of hypothalamic centers to estrogenic regulation may be impaired with age and contribute to the loss of reproductive function in female rats. Here, we have tested the hypothesis that aging is associated with alterations in the level of expression of the estrogen receptor (ER) gene and/or the neurotensin/neuromedin-N (NT/N) gene in the preoptic area (POA) of female rats. We have used in situ hybridization histochemistry and quantitative autoradiography to compare ER gene expression and NT/N gene expression in the POA of ovariectomized and ovariectomized/estradiol-treated female rats at 3, 11, and 20 months of age. We found no evidence for an age-related impairment of either ER or NT/N gene expression in two subdivisions of the POA: the anterior medial preoptic nucleus and the medial preoptic nucleus. Likewise, estrogenic regulation of both ER messenger RNA levels and NT/N messenger RNA levels did not differ across age groups. These results indicate that transcription of the ER gene within the POA is not reduced with age and suggest that the receptor translated within the POA functions normally in old female rats. Our observations do not support a role for impaired expression of the ER gene or impaired estrogenic induction of NT/N gene expression by preoptic neurons in the development of reproductive acyclicity with aging.

Aging↗
PubMed1994

Comparison of calcium, magnesium-ATPase activity and membrane fluidity in patients with essential hypertension and in normotensive controls.

OBJECTIVES: To measure calcium,magnesium-ATPase (Ca-ATPase) activity and membrane fluidity in patients with essential hypertension compared with normotensive subjects; to investigate the interrelationship between membrane fluidity and the activity of the Ca-ATPase; and to assess the importance of circulating lipids on the Ca-ATPase and membrane fluidity. METHODS: Ca-ATPase and membrane fluidity were measured in erythrocyte membranes. Kinetic parameters [maximal activity (Vmax), apparent dissociation constant and allosteric number] of the Ca-ATPase activity were measured, in the presence of saturating calmodulin, in 38 normotensives and 57 essential hypertensives. Fluorescent polarization anisotropy, as an index of membrane fluidity, was measured, using the fluorescent probes 1,6-diphenyl-1,3,5-hexatriene (DPH) and trimethylammonium DPH (TMA-DPH) in 37 normotensives and 44 hypertensives. Of these 22 were paired for age, sex and race. RESULTS: There was no significant difference in the Vmax and allosteric number of the Ca-ATPase between the normotensives and hypertensives, but there was a trend for the hypertensives to have a reduced calcium affinity. In contrast, hypertensive subjects had significantly lower membrane fluidity. Sex and serum triglycerides level were important determinants of membrane fluidity in both groups. Comparisons between normotensives and hypertensives demonstrated decreased fluidity in the hypertensives independent of sex and serum triglycerides level, although the differences, especially with TMA-DPH, were more pronounced in the females. In both groups there were negative correlations between Vmax and both DPH and TMA-DPH anisotropy. CONCLUSION: The present study demonstrates that essential hypertension is associated with a generalized alteration in the erythrocyte membrane physical and chemical properties. However, despite the positive correlation between Vmax and membrane fluidity, the present study also demonstrates that essential hypertension is not associated with a major abnormality in the activity of the erythrocyte Ca-ATPase in isolated membranes.

Adult↗
PubMed1994

Hypertension-related coronary thrombosis: prothrombic role of angiotensin II.

Although hypertension is a major risk factor in acute myocardial infarction, concomitant hypercoagulability causing thrombosis leading to myocardial infarction remains unproven for lack of an appropriate coagulation test. This study was devised to determine whether a modified recalcification time (MRT) test can demonstrate that angiotensin II, a potent vasoconstrictor, also accelerates coagulation to promote thrombosis. The MRT incorporates blood cells and chemical coagulants for maximizing sensitivity. Four groups (A, B, C, and D) of aliquots of citrated human blood were incubated for 2 hours at 37 degrees C after adding to A--20 microL saline, to B--10 micrograms Escherichia coli endotoxin, to C--20 micrograms angiotensin II, and to D--a combination of E coli endotoxin and angiotensin II. The experiment was repeated with nonincubated aliquots. Modified recalcification time values +/- standard deviation in minutes were: A--5.5 +/- 1.5, B--4.6 +/- 1.1, C--4.9 +/- 1.0, and D--3.9 +/- 1.0. Significance (Student's t test) was as follows: B versus A P < .001; C versus A, P < .05; C versus D, P < .001; B versus C, P < .05; and B versus D, P < .001. No significant changes occurred in nonincubated blood. We conclude that angiotensin II has a hypercoagulable effect, as does endotoxin. The hypercoagulability in concert with vasospasm can explain the role of hypertension in acute myocardial infarction. This in vitro study excludes the role of other in vivo mechanisms in the development of angiotensin II-induced hypercoagulability.

Angiotensin II↗
PubMed1994

Erythrocyte sodium-lithium countertransport: relationship with plasma renin activity, aldosterone and atrial natriuretic peptide in a population study.

The objective of this study was to investigate the relationships between erythrocyte Na-Li countertransport, plasma renin activity (PRA), plasma aldosterone and plasma atrial natriuretic peptide (ANP) in a population-based study, in a cross-sectional observation of a randomly selected sample of a male working population, conducted at the Medical Care Center at the Olivetti Factory in Pozzuoli (Naples). One hundred and seventy-nine male employees aged 45.3 +/- 6.8 years (mean +/- SD), all on unrestricted diet and on no pharmacological treatment were studied. Statistical associations between Na-Li countertransport activity and the hormonal factors being studied were investigated, with control for the possible confounding effects of age, BP and sodium intake. Na-Li countertransport was significantly and positively associated with plasma aldosterone concentration (r = 0.228, P < 0.01) but not with PRA or plasma ANP. Men in the highest quartile of the Na-Li countertransport distribution had significantly higher plasma aldosterone levels than men in the other quartiles (P < 0.01) and this difference was independent of age, body mass, PRA, urinary sodium excretion and blood pressure. It is concluded that plasma aldosterone levels are enhanced and disproportionately high relative to plasma renin activity in individuals with high Na-Li countertransport, independently of known potential confounders.

Adult↗
PubMed1994

Reactivity of polyclonal human CD3 antiserum in lymphoid tissues of cattle, sheep, goats, rats and mice.

Polyclonal rabbit antiserum to human T-cell CD3 was used to study its reactivity in lymphoid tissues (lymph nodes, spleen, aggregated lymphoid follicles [Peyer's patches], thymus) of several animal species (cattle, sheep, goats, rats, and mice). Using a peroxidase-antiperoxidase technique on formalin-fixed and paraffin-embedded tissues, immunoreactive cells were detected in T cell-dependent areas of the lymphoid tissues. Reactivity was high in all species tested, but mouse tissues had reduced reactivity, compared with the other species. To obtain a reaction, it was necessary to digest tissues with pronase before application of the immunocytochemical technique. Our results indicate that CD3 antiserum may specifically recognize T-lymphoid cells as it does in human lymphoid tissues and can be used as a marker to study physiologic and pathologic conditions of the lymphoid system of these species.

Animals↗
PubMed1993

Seasonal flank alopecia in boxers and Airedale terriers: 24 cases (1985-1992).

Clinical and histologic features of seasonal flank alopecia in 12 Airedale Terriers and 12 Boxers were reviewed. Most of the affected dogs were spayed females; however, sexually intact females as well as sexually intact and neutered male dogs with the disease were identified. Mean (+/- SD) age of onset was 3.6 +/- 2.4 years.

Alopecia↗
PubMed1993

Histidine 52 is a critical residue for rapid formation of cytochrome c peroxidase compound I.

The crystal structure and reactivity with hydrogen peroxide are reported for a mutant of a cloned cytochrome c peroxidase [CcP(MI)], in which the conserved distal His (His-52) is replaced with Leu. The reaction of the H52L enzyme with peroxide was examined as a function of pH in 0.1 M phosphate buffers and buffers in which nitrate was used to adjust the ionic strength. The pH-independent bimolecular rate constant for the reaction of H52L with peroxide was 731 +/- 44 and 236 +/- 14 M-1 s-1 in phosphate and nitrate-containing buffers, respectively. This represents a 10(5)-fold decrease in rate relative to the CcP(MI) parent under comparable conditions. Single-crystal diffraction studies showed that no dramatic changes in the structure or in the accessibility of the heme binding site were caused by the mutation. Rather, the mutation caused significant structural changes only at residue 52 and the nearby active-site water molecules. The residual reactivity of the H52L enzyme with peroxide was pH- and buffer-dependent. In nitrate-containing buffer, the apparent bimolecular rate constant for the reaction with peroxide decreased with decreasing pH; the loss of reactivity correlated with protonation of a group with an apparent pKA = 4.5. Protonation of the group caused a loss of reactivity with peroxide. This is in contrast to the CcP(MI) parent enzyme, as well as all other mutants that have been examined, where the loss of reactivity correlates with protonation of an enzyme group with an apparent pKA = 5.4.(ABSTRACT TRUNCATED AT 250 WORDS)

Crystallography↗
PubMed1993

Effect of arginine-48 replacement on the reaction between cytochrome c peroxidase and hydrogen peroxide.

The crystallographic structures of two cytochrome c peroxidase (CcP) mutants, CcP(R48L) and CcP(R48K), have been determined. In addition, the electronic absorption spectrum and the hydrogen peroxide reactivity of these two mutants have been determined between pH 4 and 8. Both the crystallographic structure and the electronic absorption spectrum of CcP(R48L) are consistent with exclusive pentacoordination of the heme iron between pH 4 and 6.5. At higher pH, CcP(R48L) forms an alkaline bis-imidazole form of CcP with the distal histidine coordinated to the heme iron. The apparent pKA for this transition is 7.5 in CcP(R48L). The observed pseudo-first-order rate constant for the reaction between CcP(R48L) and hydrogen peroxide saturates at high peroxide concentrations. The data are consistent with a rate-limiting oxygen-oxygen bond scission at high peroxide concentrations. The observed rate of the bond scission step ranges between 1000 and 1950 s-1, an estimated 2 orders of magnitude slower than for wild-type enzyme. The data suggest that the protonated form of His-52 increases the bond scission step by a factor of 2. The properties of the CcP(R48K) mutant are significantly different from those of CcP(R48L). The crystal structure of CcP(R48K) shows Lys-48 occupying the putative peroxide binding site. The electronic absorption spectrum indicates that CcP(R48K) is predominantly pentacoordinate at neutral pH but with detectable amounts of hexacoordinate forms. Two ionizable groups affect the electronic absorption spectrum of CcP(R48K). An apparent ionization near pH 4 produces an enzyme with increased hexacoordination, while an apparent pKA of 6.9 generates the alkaline bis-imidazole form. The peroxide reaction saturates at high peroxide concentrations for CcP(R48K) and is attributed to a conformational-gating mechanism. The maximum rate for the reaction between CcP(R48K) and hydrogen peroxide is probably limited by the movement of either Lys-48 or His-52. This rate is 200 and 290 s-1 in nitrate-containing buffers and phosphate buffers, respectively. Evidence is provided that Arg-48 in wild-type enzyme is responsible for nitrate binding in the heme pocket and for stabilizing CcP Compound I.

Arginine↗
PubMed1993

Testosterone regulates galanin gene expression in the bed nucleus of the stria terminalis.

The regulation of galanin (GAL) gene expression in the bed nucleus of the stria terminalis (BNST) by testosterone (T) was investigated using in situ hybridization histochemistry. Castration of adult male rats significantly reduced both the number of cells which expressed GAL mRNA and the average number of grains per cell. These effects were reversed by testosterone treatment. Testosterone stimulates GAL gene expression in the same neurons that have previously been shown to exhibit steroid regulation of vasopressin gene expression.

Animals↗