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Biomedical subjects

K D Pettigrew

Publications and source records attributed to K D Pettigrew.

At least 37 records · Page 2Linked to original sources

Effects of arginine vasopressin and atriopeptin on glial cell volume measured as 3-MG space.

This study evaluates the hypothesis that arginine vasopressin (AVP) and atriopeptin, peptide hormones synthesized and released within the brain, are regulators of brain cell volume using cultured astroglial cells derived from newborn rats. Cell water content, regarded as volume, was measured in defined, serum-free medium as the 3-O-methylglucose (3-MG) space. Initial experiments established conditions such that glucose, which competes with 3-MG for the glucose carrier, would not interfere with the measurement of the 3-MG space. AVP increased the 3-MG space of glial cells by an average of 25% between 30 and 120 min of exposure, whereas atriopeptin decreased it by 32%. The 3-MG space remained close to normal after coadministration of both peptides. The AVP-dependent increase in 3-MG space was blocked both by the V1 antagonist d(CH2)5Tyr(Me)AVP (Manning compound) and by the cotransport inhibitor, bumetanide. Results are consistent with a role for AVP and atriopeptin in the homeostasis of atroglial cell volume.

3-O-Methylglucose↗

Transient impairment of recognition memory following ibotenic-acid lesions of the basal forebrain in macaques.

To assess the contributions of the basal forebrain cholinergic nuclei to visual recognition memory in macaques, we compared the effects of lesions of (a) the nucleus basalis of Meynert, (b) the medial septal and diagonal band nuclei, and (c) all nuclei combined on performance of delayed nonmatching-to-sample with trial-unique stimuli. Whereas monkeys with the separate lesions did not differ from each other or from normal control animals, those with combined lesions showed a significant impairment. With time and extended practice, however, the performance of the animals with combined lesions recovered to normal levels. During the recovery period, these monkeys showed an initially increased sensitivity to scopolamine that later dissipated, at which time they also failed to show the improvement that follows physostigmine administration in normal animals. Postmortem assessment of cortical choline acetyltransferase activity revealed that only the group with combined lesions had significant depletion of this enzyme. The results suggest that (1) the basal forebrain cholinergic system participates in mnemonic processes in primates and that (2) extensive damage to this system is necessary before impairments in recognition memory, even transient ones, can be observed.

Acetylcholinesterase↗

Extracellular volume decreases while cell volume is maintained by ion uptake in rat brain during acute hypernatremia.

1. Regulation of brain extracellular and intracellular water content, regarded as volume, and electrolytes in response to 90 min of hypernatremia has been studied in the cerebral cortex of rats under urethane anaesthetic. 2. Total tissue electrolytes and water were partitioned between extracellular and intracellular compartments based on measurements made in two series of experiments. In one, tissue samples were collected and analysed for total water, Na+, K+ and Cl-. In the other, tissue extracellular volume fraction, [Na+] and [K+] were measured in situ using ion-selective microelectrodes. 3. Osmotically induced water loss from cerebral cortex was less than that predicted for ideal osmotic behaviour, revealing a degree of volume regulation, and this regulation was associated with net tissue uptake of Na+, Cl- and K+. 4. Total water content was 3.77 g H2O (g dry weight)-1 in control cortex and this decreased by 7% after 30 min of hypernatremia and then remained relatively stable at this value. Control extracellular water content, based on an extracellular volume fraction of 0.18, was 0.88 g H2O (g dry weight)-1. Control intracellular water content, estimated as the difference between total and extracellular water contents, was 2.89 g H2O (g dry weight)-1. After 30 min of hypernatremia, extracellular water content decreased by an average of 27% but intracellular water did not change. This indicates selective regulation of cell volume. By 90 min the extracellular water content had decreased by 47% and the loss in extracellular water content appeared to be accompanied by a roughly equivalent increase in intracellular water content. The intracellular volume increase, however, was not statistically significant. The tortuosity of the extracellular space averaged 1.57 and increased to 1.65 during the hypernatremia. 5. Brain extracellular fluid and plasma [Na+] were roughly equal in control tissue. Both increased by 30 mu equiv (g H2O)-1 as a result of the hypernatremia, although extracellular [Na+] lagged behind the plasma value during much of the first 60 min of hypernatremia. Extracellular [K+] was homeostatically regulated at 3 mu equiv (g H2O)-1 independent of changes in plasma electrolytes. 6. Estimates of extracellular and intracellular ion content (mu equiv (g dry weight)-1) indicate that extracellular Na+, Cl- and K+ content decreased during hypernatremia, by 32, 21 and 42% respectively, whereas intracellular ion content increased by 100, 169 and 5% respectively. 7. It is concluded that during acute hypernatremia the extracellular space decreases in volume through the loss of water and electrolytes while the intracellular compartment maintains its water content and gains electrolytes.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Variation in local cerebral blood flow response to high-dose pentobarbital sodium in the rat.

Microvascular bed structure and functions are known to vary throughout the brain. Microvascular responses to high doses of pentobarbital sodium might therefore differ among brain areas. This possibility was examined by measuring local cerebral blood flow (LCBF) with [14C]iodoantipyrine in 52 brain areas at 5, 10, 25, and 60 min after intraperitoneal administration of pentobarbital (50 mg/kg). From 5 to 60 min, LCBF was significantly lowered in 17 of 25 forebrain gray matter areas but in only 1 of 18 hindbrain gray matter structures, the pontine nuclei. Smaller, shorter duration lowering of LCBF was also observed in ten other brain areas. In both control and treated rats, LCBF was found to vary within individual brain structures. The pattern of these LCBF variations was columnar in the cerebral cortex and the hippocampus but was patchy in the caudate-putamen, thalamus, and inferior colliculus. These results indicate that pentobarbital anesthesia more strongly alters LCBF in the forebrain than in the hindbrain and produces different patterns of changes in LCBF than in local cerebral glucose utilization, which was measured with 2-deoxyglucose in a companion study.

Animals↗

Discordance and concordance of dementia of the Alzheimer type (DAT) in monozygotic twins indicate heritable and sporadic forms of Alzheimer's disease.

Monozygotic (MZ) twin pairs concordant for dementia of the Alzheimer type (DAT) or for proven Alzheimer's disease (AD) have a significantly higher frequency of a positive family history (DAT or AD in at least one first-degree relative, p less than 0.002) than do discordant MZ twin pairs, consistent with a lesser predicted distribution of proportions of surviving first-degree relatives without DAT (or AD) (p less than 0.001). The results suggest that concordant MZ twin pairs with DAT (or AD) have a heritable form of disease. AD in discordant twins may result from environmental influences or from a somatic chromosomal change following zygotic division.

Adult↗

Regrowth of damaged neurosecretory axons to fenestrated vessels of implanted peripheral tissues.

A major target of neurosecretory axons (NSA) is the basal lamina around fenestrated blood vessels (FBV) in the neural lobe of the pituitary gland. We have posed the question of whether there is neurovascular specificity. Do mature, regenerating NSA terminate selectively on the FBV of the neural lobe compared with the FBV of other tissues that normally are not innervated by NSA? Three types of tissue were transplanted between inbred Fisher rats. Fragments, about 1 mm3, of pineal, adrenal medulla, and neural lobe were grafted bilaterally to the hypothalamic, retro-chiasmatic area, which includes bundles of NSA from supraoptic and paraventricular neurosecretory nuclei exclusively, but no FBV. Two and 4 weeks later, the grafts were prepared for the immunohistochemical localization of NSA and for electron microscopy. NSA-FBV proximity was measured, and the number of NSA, FBV, and of NSA-FBV associations was counted per surface area of each graft. Regenerating NSA can associate as closely with FBV of other tissues as they can with the FBV of the neural lobe. There does not appear to be specificity with respect to the closeness of association between neurosecretory terminals and fenestrated capillaries. However, the number of these associations is greater in neural lobe grafts than in adrenal or pineal grafts at 4 weeks. The number of FBV is also greatest in neural lobe grafts at this time, an increase that would provide a greater opportunity for NSA-FBV associations.

Adrenal Medulla↗

Kinetic analysis of blood-brain barrier transport of D-glucose in man: quantitative evaluation in the presence of tracer backflux and capillary heterogeneity.

The present study deals with the analysis of double-indicator curves for blood-brain barrier studies. Two mathematical models which provide for the estimation of backflux of tracer from brain to blood in conjunction with heterogeneity of the cerebral capillary and large-vessel transit times were used for the analysis of D-glucose transport on the basis of cerebral venous outflow curves. The two models, non-mixed and well mixed, arise from differing assumptions regarding the effective region surrounding the capillary lumen. An approximate solution for the well-mixed model was developed to increase computation speed. Fourteen D-glucose outflow curves and their reference curves were obtained from nine patients and subsequently analyzed by the two models. Further, in five patients data were obtained under different physiological conditions: normal, decreased, and increased cerebral blood flow rates. The results support the appropriateness of the well-mixed model and heterogeneity of the cerebral capillary transit times. The median value for the average extraction was 0.18 and the median distribution space was 0.14. The latter value is similar to the brain extracellular space that has been estimated by other methods. The extraction values calculated from the peak of the venous outflow curves were significantly smaller than the whole-brain average extraction values estimated with the well-mixed model (0.157 vs 0.178, P less than 0.0005). In summary: (a) capillary heterogeneity is present in the human brain and changes with cerebral blood flow; (b) after crossing the blood-brain barrier, D-glucose distributes in the brain extracellular fluid; and (c) the extraction curve is significantly influenced by backflux.

Biological Transport, Active↗

Asymmetrical transport of amino acids across the blood-brain barrier in humans.

Blood-brain barrier permeability to four large neutral and one basic amino acid was studied in 30 patients with the double indicator technique. The resultant 64 venous outflow curves were analyzed by means of two models that take tracer backflux and capillary heterogeneity into account. The first model considers the blood-brain barrier as a double membrane where amino acids from plasma enter the endothelial cell. When an endothelial cell volume of 0.001 ml/g was assumed, permeability from the blood into the endothelial cell was, for most amino acids, about 10-20 times larger than the permeability for the reverse direction. The second model assumes that the amino acids, after intracarotid injection, cross a single membrane barrier and enter a well-mixed compartment, the brain extracellular fluid, i.e., the endothelial cell is assumed to behave as a single membrane. With this model, for large neutral amino acids, the permeability out of the extracellular fluid space back to the blood was between 8 to 12 times higher than the permeability from the blood into the brain. Such a difference in permeabilities across the blood-brain barrier can almost entirely be ascribed to the effect of a nonlinear transport system combined with a relatively small brain amino acid metabolism. The significance of the possible presence of an energy-dependent A system at the abluminal side of the blood-brain barrier is discussed and related to the present findings. For both models, calculation of brain extraction by simple peak extraction values underestimates true unidirectional brain uptake by 17-40%. This raises methodological problems when estimating blood to brain transfer of amino acids with this traditional in vivo method.

Adult↗

Cerebrovascular transport of [125I]quinuclidinyl benzilate, [3H]cyclofoxy, and [14C]iodoantipyrine.

The transport rate constants across rat brain capillaries of a muscarinic acetylcholine receptor antagonist, [125I]quinuclidinyl benzilate (IQNB), an opiate receptor antagonist, [3H]cyclofoxy (CF), and a highly diffusible blood flow indicator, [14C]iodoantipyrine (IAP), were determined by the indicator-diffusion technique and a model that includes a distribution of capillary transit times and a homogeneous distribution of the test compound in the tissue parenchyma. The mean influx extraction ratio (E1) of IAP was greater than 0.91, and E1 for CF and IQNB was 0.56-0.79 and 0.34-0.46, respectively. The order of lipid solubility was IQNB greater than IAP greater than CF; the apparent discrepancy (high lipid solubility and low permeability) of IQNB was partly due to intravascular binding to plasma protein and red blood cells. The observed initial tissue distribution volumes (lambda 1, ml/g brain) for IQNB (0.09-0.17), CF (0.51), and IAP (0.71) were compared with those estimated for the unbound free ligand in blood (lambda a, ml/g brain; IQNB = 1.3-2.3, CF = 0.88, and IAP = 1.4). These findings suggest that the binding of lipid-soluble radioligands and drugs to plasma proteins and red blood cells can be a major determinant of transport across the blood-brain barrier and the apparent distribution volume of the ligand in brain tissue.

Animals↗

Cerebral vessels cryofixed after hyperosmosis or cold injury in normothermic and hypothermic frogs.

Three purported means by which large solutes may penetrate the blood-brain barrier are: permeabilized tight junctions; vesicular transport; or channel formation across cerebral blood vessels. The role of vesicular transport has been questioned, in part, because many cytoplasmic vesicles are induced by aldehyde fixation. Cryofixation reduces this artefact and was used to see structural changes in frog cerebral endothelium made permeable to plasma solutes after perivascular exposure to hyperosmotic (3 M) urea, or injury with a cold probe (-50 degrees C). Some control and experimental frogs were made hypothermic so as to inhibit endocytosis and autolytic changes. The brains of some untreated controls were immerse-fixed in aldehydes. Other controls and all other brains of normothermic or hypothermic animals were rapidly frozen, then substituted with acetone-fixative. The interendothelial tight junctions separate partially or completely, after hyperosmotic exposure, in one third of the junctions. Blood-borne ferritin and Evans blue pass through some of the patent junctions. Junctional opening is caused by cell shrinkage, because the perimeter/area ratio of individual endothelial cells in the hyperosmotic group is significantly greater than in the control, due to a decreased area. Large 0.08-0.32-micron-wide invaginations or pits of the endothelial cell membrane characterize both cryofixed and aldehyde-fixed vessels. The pits often appear as isolated vesicles in the cytoplasm, but serial sections reveal that many communicate with either the capillary lumen or subendothelial space. No series of pits opened onto both lumen and space to form a transendothelial channel. The number of vesicles in aldehyde-fixed specimens is about 4 times greater (P less than 0.01) and in the cold injured, cryofixed brain capillary, about two times greater (P less than 0.01), than in the cryofixed control. Hyperosmotic exposure does not increase the number of pits. The permeabilization of anuran cerebral endothelium by hyperosmotic treatment or cold injury is thus by means of an intercellular rather than a transcellular route.

Animals↗

Dexamethasone effects on vascular volume and tissue hematocrit in experimental RG-2 gliomas and adjacent brain.

We have studied the effects of dexamethasone, a corticosteroid commonly used to treat brain tumors, on vascular volume and tissue hematocrit in RG-2 experimental rat gliomas. 125I-RISA (radioiodinated serum albumin) was used to measure tissue plasma vascular volume (Vp) and 51Cr labeled red cells were used to measure tissue red cell volume (Vrbc). Quantitative autoradiography was used to obtain local measurements of Vp and Vrbc in different brain and tumor regions. From these experimentally measured values, we calculated the tissue vascular volume (Vv), tissue hematocrit (THct) and systemic arterial hematocrit (AHct). The value reported primarily reflect capillary and small vessel volumes since blood drained from larger vessels during tissue processing and large vascular structures were avoided during analysis of the autoradiographic images. A total of 110 tumors were studied in 29 animals. There was a consistent trend for Vp and Vv to be reduced in all tumor regions after dexamethasone treatment, although a significant decrease was seen only in tumor center. Dexamethasone did not affect Vp or Vv in tumor-free brain regions. Dexamethasone appeared to have little effect on Vrbc in any brain or tumor region. THct was consistently, although not significantly, higher in tumors after treatment with dexamethasone; THct in tumor-free brain regions was unaffected by dexamethasone treatment.

Animals↗

Motor activity and sleep duration as a function of age in healthy men.

Patient activity monitors (PAM's) and self-report diaries were employed to examine wrist movements over a 9 day period in 43 healthy men aged 21 to 83 years, in their natural home and work environments. The mean sleep duration (from PAM) equaled 7.29 +/- 0.8 (S.D.) hr compared to 7.84 +/- 0.62 hr as estimated from the daily diaries. Neither parameter was correlated significantly with age (p greater than 0.05). Net activity counts per 24 hr day declined significantly with age (p less than 0.05), but the effect of age was small and accounted for only 15% of the variance. When the data from each age group were averaged into mean 24 hr days, it was found that: (1) the greatest reductions in the older subjects' activity occurred during evening hours; (2) differences between old and young subjects were more profound on weekends than on weekdays; and (3) only the young subjects' activity profiles differed between weekdays and weekends, as they rose from bed later on weekends. The results indicate that 24 hr activity can be quantified and characterized in natural work and home environments, in relation to diary information. Only small age differences occur in overall 24 hr activity in rigorously screened healthy men.

Adult↗

Rapid dynamic CT scanning to distinguish schizophrenic from normal subjects.

Rapid dynamic computerized axial tomography (CT) density-time curves were used to detect abnormal brain regions in groups of controls and DSM-III diagnosed schizophrenics. Density-time curves were designated as plateau curves if, following a bolus injection of contrast material, they exhibited a plateau from the peak value rather than a decrease. Five of the 10 schizophrenics versus none of the 11 controls were found to possess two or more plateau curves. The physiological basis for the plateau curves is not known, but an increase in blood-brain-barrier permeability could be involved.

Adult↗

Volume regulatory loss of Na, Cl, and K from rat brain during acute hyponatremia.

This study quantitatively evaluates the contribution of tissue Na, Cl, and K loss to brain volume regulation during acute dilutional hyponatremia (DH) and examines the mechanism of Na loss. DH was produced in pentobarbital sodium-anesthetized rats by intraperitoneal infusion of distilled water and brain water and electrolytes analyzed 30 min, 1 h, 3 h, 4 h, or 6 h later. The rate of Na and Cl loss was greatest during the first 30 min of DH (0.43 and 0.47 meq X kg tissue dry wt-1 X min-1, respectively). Net loss of Na and Cl was maximal after 3 h of DH. K loss was slower, achieving significance after 3 h. Electrolyte loss was sufficient to account for observed brain volume regulation after three or more hours of DH. Measurements of 22Na influx and efflux across the blood-brain barrier showed that barrier permeability to Na is unchanged during DH. Analysis of results using a two-compartment model of plasma-brain exchange suggests that loss of brain Na during DH does not result solely from a shift of electrolyte across the blood-brain barrier to plasma, and thus provides indirect evidence for an additional pathway for Na loss, presumably via cerebrospinal fluid.

Animals↗

Apparent glucose utilization in Walker 256 metastatic brain tumors.

Regional rates of apparent glucose utilization (GU) in metastatic Walker 256 (WL-256) brain tumors produced by the intracarotid injection of WL-256 tumor cells in rats were measured using 14C-deoxyglucose and quantitative autoradiography. Apparent glucose utilization was uniform within individual small and medium size tumors without necrosis, varied considerably among different tumors within this group, and did not correlate with tumor size or location. High values of GU in medium and large-size tumors correlated with viable-appearing tissue in contrast to necrotic tissue and were always 1.3 to 3 times higher than that of adjacent and contralateral nontumorous brain. The apparent net extraction of glucose (En) in viable tumor regions was estimated to be several fold higher than that in remote brain tissue; analysis of this data for medium and large tumors indicates that the calculated values of GU and En overestimate the actual rates of utilization and net extraction of glucose. Local cerebral glucose utilization (LCGU) was higher than normal adjacent to small tumors and lower than normal adjacent to large tumors. The LCGU in many gray-matter structures remote from the intracerebral tumors was reduced and roughly proportional to the metastatic tumor burden. The comparatively high uptake of 2-deoxyglucose by viable tumor cells has diagnostic and localization value and suggests that appropriate glucose analogues could be developed to produce a tumor-selective inhibition of glycolysis and tumoricidal effect.

Animals↗

Apparent glucose utilization in Walker 256 metastatic brain tumors.

Regional rates of apparent glucose utilization (GU) in metastatic Walker 256 (WL-256) brain tumors produced by the intracarotid injection of WL-256 tumor cells in rats were measured using 14C-deoxyglucose and quantitative autoradiography. Apparent glucose utilization was uniform within individual small and medium size tumors without necrosis, varied considerably among different tumors within this group, and did not correlate with tumor size or location. High values of GU in medium and large-size tumors correlated with viable-appearing tissue in contrast to necrotic tissue and were always 1.3 to 3 times higher than that of adjacent and contralateral nontumorous brain. The apparent net extraction of glucose (En*) in viable tumor regions was estimated to be several fold higher than that in remote brain tissue; analysis of this data for medium and large tumors indicates that the calculated values of GU and En* overestimate the actual rates of utilization and net extraction of glucose. Local cerebral glucose utilization (LCGU) was higher than normal adjacent to small tumors and lower than normal adjacent to larger tumors. The LCGU in many gray-matter structures remote from the intracerebral tumors was reduced and roughly proportional to the metastatic tumor burden. The comparatively high uptake of 2-deoxyglucose by viable tumor cells has diagnostic value and suggests that appropriate glucose analogues could be developed to produce a tumor-selective inhibition of glycolysis and tumoricidal effect.

Animals↗

The natural history of homocystinuria due to cystathionine beta-synthase deficiency.

An international questionnaire survey has been conducted to define better the natural history of homocystinuria due to cystathionine beta-synthase deficiency and permit evaluation of treatment. Data were compiled for 629 patients. Among patients not discovered by newborn screening, B6-responsive individuals on the average have significantly better mental capabilities (mean IQ, 79) than do B6-nonresponsive individuals (mean IQ, 57). Time-to-event curves are presented for the other major clinical abnormalities produced by this disease. Each occurred at significantly lower rates in untreated B6-responsive than in untreated B6-nonresponsive patients, as shown by the following examples: (1) dislocation of optic lenses (at age 10, chances of dislocation: 55% and 82%, respectively); (2) initial clinically detected thromboembolic events (at age 15, chances of having had such an event: 12% and 27%, respectively); (3) radiologic detection of spinal osteoporosis (at age 15, chances of such osteoporosis having been detected: 36% and 64%, respectively); and (4) mortality (at age 30, chances of not surviving: 4% and 23%, respectively). Methionine restriction initiated neonatally prevented mental retardation, retarded the rate of lens dislocation, and may have reduced the incidence of seizures. Pyridoxine treatment of late-detected B6-responsive patients retarded the rate of occurrence of initial thromboembolic events. Following 586 surgical procedures, 25 postoperative thromboembolic complications occurred, six of which were fatal. Reproductive histories were reported predominantly for B6-responsive patients. Living offspring of either men or women patients had few abnormalities. The evidence is inconclusive whether untreated maternal cystathionine beta-synthase deficiency leads to excessive fetal loss. Only 13% of patients detected in screening programs of newborns and classified as to B6-responsiveness were B6-responsive, compared to 47% among late-detected patients. Current screening programs that identify neonatal hypermethioninemia may be preferentially failing to detect B6-responsive patients.

Adolescent↗

Local cerebral glucose utilization in fetal and neonatal sheep.

The newborn mammalian brain of several species has been shown to have a lower average rate of energy metabolism and a narrower range of rates in its various components than is found in maturity. In a further study of cerebral energy metabolism during development, we have employed the [14C]deoxyglucose method for measuring local cerebral glucose utilization in fetal and neonatal sheep. After establishing the lumped constant to be 0.40 and finding the rate constants for the kinetic behavior of deoxyglucose in plasma and brain to be close to those in other species, we measured the rates of glucose utilization in 44 regions of the brain. The rates were low and homogeneous in midgestation, except for those of brain stem nuclei of the auditory and vestibular systems and those of the hippocampus which were relatively high. In the last 7 wk, local rates rose approximately threefold. After birth there was a further average increase of 50% above full-term levels. The study shows that cerebral energy metabolism rises in most structures during prenatal maturation, a time when sensory stimulation is at a relatively low level and behavioral responses are minimal.

Animals↗