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Biomedical subjects

R F Keep

Publications and source records attributed to R F Keep.

At least 91 records · Page 5Linked to original sources
PubMed1994

Developmental stages in experimental liver metastases: relation to invasiveness.

We have previously reported that an invasive morphotype can be evoked in a rat colon carcinoma by transplanting it into pre-induced subcutaneous granulation tissue. We have now studied the interaction of the same tumor with liver tissue, which is extremely poor in connective tissue in comparison with the subcutaneous site. Tumor cells were injected into the portal system and the resulting experimental liver metastases were examined by electron microscopy and immunohistochemistry. Early metastases consisted of well-differentiated acini, fully surrounded by connective tissue that was derived from the periportal stroma. In a later stage, this connective tissue was overgrown by tumor cells and, almost immediately, acinar differentiation was lost. Most metastases eventually reached the liver capsule, which reacted by forming a layer of granulation tissue. Only in this layer, we observed invasion by thin tumor cell strands, which were often intimately associated with fibroblasts or with blood capillaries. The tumor cells remained smooth and rounded during this process. After fully penetrating the granulation tissue, the tumor cell strands reached the liver surface, where they formed poorly structured papillary masses that were nearly devoid of stroma. Our observations indicate that, even in a relatively homogeneous organ like the liver, the tumor-host interaction is highly complex and dynamic. They also confirm the notion that granulation tissue stimulates tumor invasiveness. Finally, they show that tumor cells can actively invade host tissues without exhibiting a "fibroblastic" morphology.

Animals↗
PubMed1994

Blood-brain barrier permeability and brain concentration of sodium, potassium, and chloride during focal ischemia.

Brain edema formation during the early stages of focal cerebral ischemia is associated with an increase in both sodium content and blood-brain barrier (BBB) sodium transport. The goals of this study were to determine whether chloride is the principal anion that accumulates in ischemic brain, how the rate of BBB transport of chloride compares with its rate of accumulation, and whether the stimulation seen in BBB sodium transport is also seen with other cations. Focal ischemia was produced by occlusion of the middle cerebral artery (MCAO) in anesthetized rats. Over the first 6 h after MCAO, the amount of brain water in the center of the ischemic cortex increased progressively at a rate of 0.15 +/- 0.02 (SE) g/g dry wt/h. This was accompanied by a net increase in brain sodium (48 +/- 12 mumol/g dry wt/h) and a loss of potassium (34 +/- 7 mumol/g dry wt/h). The net rate of chloride accumulation (16 +/- 1 mumol/g dry wt/h) approximated the net rate of increase of cations. Three hours after MCAO, the BBB permeability to three ions (22Na, 36Cl, and 86Rb) and two passive permeability tracers ([3H]alpha-aminoisobutyric acid ([3H]AIB) and [14C]urea) was determined. Permeability to either passive tracer was not increased, indicating that the BBB was intact. The rate of 36Cl influx was 3 times greater and the rate of 22Na influx 1.8 times greater than their respective net rates of accumulation in ischemic brain.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Potassium cotransport at the rat choroid plexus.

The choroid plexuses are involved in cerebrospinal fluid (CSF) secretion and CSF K homeostasis. We examined K transport mechanisms present in the isolated rat choroid plexus that may be involved in these functions, predominantly using 86Rb as a marker for K. The study demonstrates that there are two primary uptake mechanisms. Ouabain-sensitive Na-K-adenosinetriphosphatase and bumetanide-sensitive cotransport, probably of the Na-K-2Cl form, account for 48 and 46% of uptake, respectively. Efflux studies demonstrate that the primary K efflux mechanism is also bumetanide-sensitive cotransport with the other major component probably being by K channels as it is inhibitable by barium or quinidine. Efflux via the cotransporter was not inhibited by R(+)-butylindazone, a KCl cotransport inhibitor, but it was enhanced in the presence of ouabain (P < 0.001) or increased extracellular Na concentration (P < 0.01). Furthermore, Na efflux was bumetanide sensitive (P < 0.05). In all, these data suggest that the efflux cotransporter is also of the Na-K-Cl form and that it is the same transporter as the influx mechanism operating in both directions. The evidence presented leads us to hypothesize that this cotransporter is on the apical membrane of the choroid plexus and that it may have a central role in CSF secretion and perhaps CSF K homeostasis.

Animals↗
PubMed1994

Rubidium entry into brain and cerebrospinal fluid during acute and chronic alterations in plasma potassium.

To elucidate whether K+ uptake across the blood-brain barrier (BBB) or blood-cerebrospinal fluid (CSF) barrier is subject to acute or chronic regulation, rats were rendered acutely or chronically hyper- or hypokalemic (range: 2.8-7.2 mM). Measurements were made of the permeability-surface (PS) products of 86Rb+, a marker for K+, and alpha-[3H]aminoisobutyric acid (AIB), a passive permeability marker, and of CSF K+ concentration ([K+]CSF). [K+]CSF decreased by 8% in chronic hypokalemia P < 0.01), but otherwise remained unchanged. The AIB PS products were unaltered in any group, excluding a change in passive permeability. The Rb PS product, however, increased by 31% for brain tissue (P < 0.01) and by 46% for CSF (P < 0.05) during acute hypokalemia, but was unchanged during acute hyperkalemia. During chronic hypokalemia the Rb PS product increased by 40% for brain (P < 0.01) and 55% for CSF (P < 0.01) and decreased during chronic hyperkalemia by 37% for brain (P < 0.01) and 49% (P < 0.01) for CSF. Unidirectional K+ fluxes were calculated, revealing greater regulation of K+ influx into both brain tissue and CSF during chronic compared with acute changes of plasma K+ concentration ([K+]pl). Our results suggest that K+ transport is saturable at both the BBB and the blood-CSF barriers under normal conditions and that both barriers adapt to chronic changes in [K+]pl by modifying specific, transcellular routes of K+ entry.

Aminoisobutyric Acids↗
PubMed1993

Transplantation of colon carcinoma into granulation tissue induces an invasive morphotype.

The stroma surrounding many malignant tumors resembles granulation tissue. To test the hypothesis that such stroma stimulates tumor invasiveness, we compared, by electron microscopy and immunohistochemistry, the growth patterns of CC531 rat colon adenocarcinoma in 2 experimental situations: (i) after transplantation into the undisturbed subcutaneous connective tissue of rats, and (ii) after transplantation into experimentally induced subcutaneous granulation tissue in rats. For the latter experimental situation, a subcutaneous "tissue chamber" was designed allowing fragments of tumor tissue to be transplanted into the very center of developing granulation tissue. In the undisturbed subcutaneous tissue, the whole tumor was generally encapsulated, and the tumor cells were arranged in compact groups with a strong tendency to form acini. In the pre-formed granulation tissue, on the other hand, the tumor tissue closely matched descriptions of invasive colon carcinomas in the literature and met the criteria for the "invasive morphotype". In this situation, the tumor consisted of thin, unorganized, widely dispersed strands of irregular tumor cells with numerous protrusions that deeply penetrated the surrounding matrix. Our results show that an invasive morphotype can be evoked by pre-inducing granulation tissue at the transplantation site.

Adenocarcinoma↗
PubMed1993

Potassium transport at the blood-brain and blood-CSF barriers.

Figure 5 gives a summary of K transporters at the BBB based on the available evidence. It appears that the cerebral endothelial cells have an array of potassium channels, although the degree to which each is open under physiological conditions is uncertain. Different channels are present on the luminal and abluminal membranes, and the opening and closing of these channels may allow modulation of the brain K influx and efflux rates and play a role in brain K homeostasis. These channels may also play a role in hyperosmotic brain volume regulation by increasing the entry rate of potassium into brain and may be involved in volume regulation of the endothelial cell itself. The nature of fluid transport at the BBB remains to be fully elucidated, with the presence of a Na/K/2Cl co-transporter being uncertain. The abluminal inwardly-rectifying channel may act as a leak pathway to allow modulation of fluid secretion by the Na/K ATPase without altering the K concentration of that fluid. Finally, there is some evidence that K transport at the BBB is under hormonal and neuronal control. The cerebral capillaries possess receptors for many of the hormones present in blood and brain.

Animals↗
PubMed1990

A morphometric study on the development of the lateral ventricle choroid plexus, choroid plexus capillaries and ventricular ependyma in the rat.

Morphometric changes in the rat lateral ventricle choroid plexus epithelium and endothelium and in the ventricular ependyma were studied between 16 days gestation and 30 days after birth, using stereological techniques. The epithelial apical surface density increased from 0.6 to 3.3 microns 2/microns 3 and the mitochondrial volume fraction from 3.2 to 7.6% during this period. The endothelial fenestrations increased from 0.05 to 0.39 micron-1. These changes may be related to postnatal increases in choroid plexus function. Morphological changes in basolateral surface density, cell height and nucleus and glycogen volume fraction have also been measured. The development of the lateral ventricle choroid plexus was qualitatively similar to the fourth ventricle plexus reported previously, but small quantitative differences occurred. The ventricular ependyma also showed a significant increase in mitochondrial volume fraction after birth, though to a lesser extent than the plexus epithelium. The total apical surface area of the choroid plexuses was estimated at 75 cm2 for 30-day-old rats. This figure, which takes into account the apical microvilli, is much greater than previous estimates and is similar to the surface area of the cerebral capillaries (155 cm2), and suggests that the choroid plexuses may play a more important role in the regulation of the brain microenvironment than previously thought.

Animals↗
PubMed1990

Cortical microvessels during brain development: a morphometric study in the rat.

In order to determine whether any structural changes occur in the blood-brain barrier during development which can be related to changes in barrier function over the same period, morphometric methods were used on pieces of visual cortex taken from rats between 16 days gestation and adult and processed for light and electron microscopy. Capillary volume fraction, numbers per unit area, and surface density increased in two phases, before birth and between 10 and 20 days after birth, with no subsequent change after 20 days. Brain parenchymal mitochondria, assumed to be a measure of brain oxidative activity, did not change before birth but increased gradually and continuously after birth from 1.7% to 5.7% in adults. The capillary endothelial mitochondrial volume fraction as a percentage of the endothelial cell cytoplasm, previously thought to be important for ion transport, did not change with age (mean = 5.4%), although there was evidence that mitochondria either divide or change in shape up to 30 days after birth. The endothelial cell thickness decreased with age from 0.6 to 0.2 microns, probably through cell elongation during vascular growth. By combining the data on endothelial mitochondria and vascularity, it was shown that the total endothelial mitochondrial volume per unit length of capillary decreased with age, whereas per unit volume of tissue it increased.

Aging↗
PubMed1990

Decrease in perfusion of cerebral capillaries during incomplete ischemia and reperfusion.

The effect of unilateral, incomplete cerebral ischemia on CBF, unidirectional flux of alpha-aminoisobutyric acid (AIB) and sodium, and number of perfused capillaries during ischemia and reperfusion was measured in the cortex of gerbils with symptomatic ischemia. Three hours of unilateral carotid occlusion reduced the CBF to the ipsilateral cortex by 81%, with a smaller 30% decrease in the contralateral cortex. Following 11 min of reperfusion, CBF in the ipsilateral cortex returned to the preischemic value, while the contralateral blood flow decreased to 50% of control. The transfer constants for AIB and sodium in the ipsilateral cortex were reduced by 67 and 53%, respectively, after 3 h of ischemia, with no change in the contralateral cortex. The transfer constant for AIB remained decreased by 48% during the first 20 min of reperfusion, while that for sodium returned to its control value. The number of perfused capillaries was reduced 54% by 3 h of ischemia and remained decreased by 20% after 11 min of reperfusion. These data indicate that 3 h of unilateral carotid occlusion reduces the number of perfused capillaries in the ipsilateral cortex during the ischemic period. Further, the early reperfusion phase is characterized by a mismatch between capillary perfusion and CBF. Finally, early in the postischemic phase, sodium transport undergoes a selective stimulation, probably as a result of stimulation of ion transport.

Aminoisobutyric Acids↗
PubMed1989

Do brain fluid ion concentrations change in hydrocephalus? A study on potassium and calcium in rats with congenital hydrocephalus.

Potassium and calcium have been measured in cerebrospinal fluid (CSF), brain interstitial fluid (ISF) and in plasma of congenitally hydrocephalic rats (H-Tx strain) and the results compared with a previous study on control, non-hydrocephalic, rats (CFY strain). Ion-selective microelectrodes were used to determine [K+] at 1-2 and 5 days and [Ca2+] at 1-2, 5, 10 and 20 days after birth. Total [Ca] was measured in plasma and CSF samples with atomic absorption spectrometry. In addition, K+ homeostasis has been studied in CSF and ISF during plasma hyperkalaemia at 5 days after birth. Apart from a raised [Ca2+] immediately after birth, hydrocephalic rats had similar CSF and ISF potassium and calcium concentrations to those found in control rats at the same age. The degree of CSF and ISF K+ homeostasis was also similar in hydrocephalic and control rats.

Animals↗
PubMed1988

Brain fluid calcium concentration and response to acute hypercalcaemia during development in the rat.

1. In vivo measurements of plasma, cerebrospinal fluid (CSF) and brain interstitial fluid (ISF) ionic Ca2+ concentrations ([Ca2+]) have been made in anaesthetized rats aged between 19 days gestation and adult using calcium-selective microelectrodes. Total calcium concentration ([ Ca]) has also been determined in plasma and CSF samples by atomic absorption spectrometry. 2. Under control conditions, plasma, CSF and ISF [Ca2+] showed a small, but significant, decrease with age. Plasma and CSF, but not ISF, showed a transient hypocalcaemia at birth. After birth there were no significant differences between plasma, CSF and ISF [Ca2+], except in adult rats where CSF [Ca2+] was significantly lower than plasma [Ca2+]. The age-related changes in CSF and ISF [Ca2+] were small and it is uncertain as to whether they may have any functional significance. 3. Under control conditions, plasma and CSF [Ca] also declined with age. The fall in plasma [Ca] paralleled the changes in plasma [Ca2+]. The decrease in CSF [Ca] was steeper than that in [Ca2+] and indicated a higher proportion of protein-bound or complexed calcium in the CSF of young when compared to old rats. 4. Acute plasma hypercalcaemia was induced by intramuscular injections of calcium gluconate and consequent changes in CSF or ISF [Ca2+] were monitored in vivo. There was very weak regulation of CSF and ISF [Ca2+] at 21 days gestation, which may reflect placental control over fetal plasma calcium. Soon after birth there was good regulation in both ISF and CSF [Ca2+]. CSF [Ca] was also measured during hypercalcaemia in samples from post-natal rats and there was similar regulation to that in CSF [Ca2+]. 5. It is concluded that under control conditions, during rat development, CSF and ISF [Ca2+] closely follow changes in plasma [Ca2+], but that soon after birth homeostatic mechanisms develop to prevent large fluctuations in brain fluid calcium.

Aging↗
PubMed1988

Brain interstitial fluid calcium concentration during development in the rat: control levels and changes in acute plasma hypercalcaemia.

Age-related changes in brain interstitial fluid (ISF) ionic calcium, in ionic and total calcium in plasma and the effect of plasma hypercalcaemia on ISF calcium have been studied in rats aged between late gestation and adult. ISF ionic [Ca2+] decreased significantly with development from 1.6 mM to 1.2 mM. Plasma ionic [Ca2+] was not significantly different from ISF [Ca2+] apart from a transient hypocalcaemia at birth which was not reflected in the ISF. Plasma total calcium was around 2X ionic [Ca2+] and showed the same age-related decrease. In acute plasma hypercalcaemia induced by calcium gluconate injections, there was only weak regulation of ISF Ca2+ at 21 days gestation but a rapid improvement after birth resulted in excellent control by 5 days.

Aging↗
PubMed1987

Choroid plexus structure and function in young rats on a high-potassium diet.

Rats, 20 days old, were fed a diet containing 20% KCl for 10 days. At 30 days these rats had lower body, brain and choroid plexus weights than matched controls and plasma [K+] was increased to 7.5 mM compared to 4.1 mM for controls. Cerebrospinal fluid (CSF) [K+] and bulk fluid secretion were not changed by the high K+ diet but there was a mild metabolic acidosis. The fourth ventricle choroid plexus was prepared for transmission electron microscopy and the structure of the epithelium analysed by quantitative stereology. The high K+ rats had a large increase in the mitochondrial volume fraction of the epithelium due to a 36% increase in individual mitochondrial volume. The high-K+ rats also had a decrease in the apical surface density of the epithelium due to a 41% decrease in the height of the microvilli. It is concluded that the increase in mitochondrial volume may provide the additional energy required for the increased transport of K+ out of the CSF in hyperkalaemia. The reduction in height of the microvilli could be a means for maintaining a normal CSF bulk secretion rate despite the increase in K+ transport required to maintain normal CSF [K+] in hyperkalaemia.

Animals↗
PubMed1987

The control of potassium concentration in the cerebrospinal fluid and brain interstitial fluid of developing rats.

1. In situ measurements of plasma, cerebrospinal fluid (CSF) and brain interstitial fluid (ISF) K+ concentrations have been made in anaesthetized rats aged between 19 days gestation and 30 days after birth using K+-selective micro-electrodes. 2. Under control conditions, plasma, CSF and ISF [K+] did not vary significantly with age, the over-all mean concentrations being 3.8, 3.1 and 3.2 mM respectively. The CSF and ISF [K+] were significantly lower than plasma [K+] at all ages. 3. It is concluded that young rats, from the age of 19 days gestation, can normally maintain plasma-to-CSF and plasma-to-ISF gradients similar to those in the adult, presumably by action of the blood-CSF and blood-brain barriers. 4. During acute plasma hyperkalaemia there was very little regulation of CSF [K+] at 21 days gestation, but after birth there was a gradual improvement in CSF K+ homeostasis until by 30 days after birth a normal CSF [K+] could be maintained. 4. There was no regulation of ISF [K+] during hyperkalaemia at 21 days gestation, although after birth there was a rapid improvement in ISF K+ homeostasis. 1-day-old neonates were able to maintain a normal forebrain ISF [K+] during hyperkalaemia. 6. It is concluded that the onset of both CSF and ISF K+ homeostasis takes place at around birth in rats. However, K+ homeostasis matures more rapidly in the ISF than in the CSF. This suggests that, at least in the neonatal period, the CSF is not essential for ISF [K+] regulation.

Animals↗
PubMed1987

Effect of chronic maternal hyperkalaemia on plasma, cerebrospinal fluid and brain interstitial fluid potassium in developing rats.

Plasma hyperkalaemia was induced in pregnant and lactating rats using a high potassium diet. Fetuses of high-K-diet mothers showed no increase in the potassium concentration [( K+]) of plasma, cerebrospinal fluid (CSF) and brain interstitial fluid, presumably due to placental control. Neonates from high-K-diet rats did show an increase in plasma [K+] but this increase was very small and there was no increase in CSF or interstitial fluid [K+]. Maternal milk [K+] was not affected by plasma hyperkalaemia. Weanling rats fed the high-K diet directly showed marked plasma hyperkalaemia but no increase in CSF or interstitial fluid [K+]. Thus, prior to weaning, a relatively stable plasma [K+] is maintained by maternal influence reducing the need for direct brain fluid K+ regulation.

Animals↗