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Biomedical subjects

P W Bankston

Publications and source records attributed to P W Bankston.

At least 19 recordsLinked to original sources
PubMed1991

Increased glucose oxidation and contents of insulin and ATP in polyamine-depleted rat insulinoma cells (RINm5F).

In order to elucidate the role of polyamines in the replication and insulin production of insulin-secreting cells, we have investigated the impact of partial polyamine depletion on the proliferation, metabolism, insulin synthesis and ultrastructure of clonal rat insulinoma cells (RINm5F). For this purpose RINm5F cells were exposed for 4 days to the specific ornithine decarboxylase (ODC) inhibitor difluoromethylornithine (DFMO). This resulted in a profound decrease in ODC activity and cytoplasmic polyamine contents. The polyamine content of cell nuclei was, however, not altered by DFMO. Addition of small amounts of putrescine during culture elevated the intracellular content of this diamine and suppressed ODC activity. The decrease in polyamine contents was accompanied by a pronounced inhibition of the cellular proliferative activity. The rates of glucose utilization, oxygen uptake and activity of the pentose cycle were decreased in DFMO-treated cells, whereas the glucose oxidation rate, oxidation/utilization ratio, ATP content and ATP/ADP ratio were increased. Insulin mRNA content and synthesis of proinsulin, insulin and total protein were not altered by DFMO. In contrast, there was a sizeable increase in the cellular insulin content, despite a lowered total protein content. Electron-microscopic analysis revealed an accumulation of insulin-secretion granules in the DFMO-treated cells. In addition, short-term insulin release was increased after DFMO exposure, but was not rendered glucose-sensitive. It is concluded that polyamines are necessary for the maintenance of rapid insulinoma-cell replication and that DFMO-treated RINm5F cells acquire an enhanced substrate oxidation and increased content of insulin and ATP.

Adenosine Diphosphate↗
PubMed1991

Differential and specific labeling of epithelial and vascular endothelial cells of the rat lung by Lycopersicon esculentum and Griffonia simplicifolia I lectins.

In the rat lung, we found that the Lycopersicon esculentum (LEA) lectin specifically binds to the epithelium of bronchioles and alveoli whereas Griffonia simplicifolia I (GS-I) binds to the endothelium of alveolar capillaries. The differential binding affinity of these lectins was examined on semithin (approximately 0.5 microns) and thin (less than 0.1 (microns) frozen sections of rat lung lavaged to remove alveolar macrophages. On semithin frozen sections, LEA bound to epithelial cells lining bronchioles and the alveoli (type I, but not type II epithelial cells). On thin frozen sections, biotinylated Lycopersicon esculentum (bLEA)-streptavidin-gold conjugates were confined primarily to the luminal plasmalemma of type I cells. bGS-I-streptavidin-Texas Red was detected on the endothelial cells of alveolar capillaries and postcapillary venules but not on those of larger venules, veins or arterioles. By electron microscopy, GS-I-streptavidin-gold complexes were localized primarily to the luminal plasmalemma of thick and thin regions of the capillary endothelium. Neither lectin labeled type II alveolar cells, but both lectins labeled macrophages in the interstitia and in incompletely lavaged alveoli.

Animals↗
PubMed1988

Measurement of live bacteria by Nomarski interference microscopy and stereologic methods as tested with macroscopic rod-shaped models.

A new method is proposed to measure bacterial cells under growth conditions. Bacterial cells, suspended in their growth medium, were attached to a cover slip with poly-L-lysine. The cover slip was inverted and placed on a glass microscope slide. To prevent dehydration of the medium, the edges of the cover slip were sealed to the microscope slide with clear fingernail polish. The bacteria on the slide were then quickly photographed with a Leitz light microscope, using Nomarski optics. The photographic negatives were then projected at a standard distance through a lens system, and the projected images of the whole cells were outlined by hand onto graph paper. The profile images so transcribed onto the graph paper were in effect transverse sections of each of the cells. Using stereologic grid and point counting techniques, the area of the cell transverse section as well as the perimeter or circumference of the transverse section were estimated. Formulae were developed so that both the volume and surface area of the whole cell could be ascertained from these area and circumference measurements. Since the efficacy of any measurements of surface area and volume of microscopic rod-shaped bacterial cells could be questioned, macroscopic rod-shaped models were used to test the theory and formulae and to compare this method with other commonly used cell-sizing techniques. This technique could be used in any study of bacterial cell size or changes in cell size (e.g., osmotic shifts).

Bacteria↗
PubMed1988

Changes in buoyant density and cell size of Escherichia coli in response to osmotic shocks.

The buoyant density of Escherichia coli was shown to be related to the osmolarity of the growth medium. This was true whether the osmolarity was adjusted with either NaCl or sucrose. When cells were grown at one osmolarity and shocked to another osmolarity, their buoyant density adjusted to nearly suit the new osmolarity. When cells were subjected to hyperosmotic shock, they became denser than expected. When cells were subjected to hypoosmotic shock they occasionally undershot the new projected density, but the undershoot was not as dramatic as the overshoot seen with hyperosmotic shocks. Shrinkage and swelling of the cells in response to osmotic shocks could account for the change in their buoyant density. The changes in cell size after osmotic shocks were measured by two independent methods. The first method measured cell size with a Coulter Counter, and the second method measured cell size by stereologic analysis of Nomarski light micrographs. Both methods gave qualitatively similar results and showed the cells to be flexible. The maximum swelling recorded was 23% of the original cell volume, while the maximum shrinkage observed was 33%.

Centrifugation, Density Gradient↗
PubMed1987

Evidence for the presence of virus in clonal insulin-producing RINm5F cells.

The ultrastructural morphology of the clonal insulin-producing cell line, RINm5F, was investigated. Virus-like particles, probably C type viruses, were identified both intra- and extracellularly. Because these particles could not be found in the original transplantable tumor, it is probable that viruses were induced at some later stage in the development of the RINm5F cell line. All investigators using the RINm5F cells should be aware of the fact that these cells may contain one or several types of viruses, and of the possibility that these particles may interfere with a variety of cellular functions.

Adenoma, Islet Cell↗
PubMed1987

Maturation of myocardial capillaries in the fetal and neonatal rat: an ultrastructural study with a morphometric analysis of the vesicle populations.

The ultrastructural morphology of the cellular and extracellular components of the developing myocardial capillary wall--from the 16-day-gestation fetus of the rat to the 21-day neonate--was examined. A morphometric analysis of plasmalemmal vesicles and of coated vesicles and pits of capillary endothelial cells was performed during the same developmental period. As the lateral extensions of the capillary endothelial cells change from irregular to regular in their thickness during development, there is an increase in the number of plasmalemmal vesicles and a progression from clusters of plasmalemmal vesicles to a uniform distribution in the endothelial cell. The ratio of vesicles which are open to the luminal front, which are "free" in the cytoplasm, or which are open to the abluminal front of the endothelial cell was consistent throughout development. The numerical density of plasmalemmal vesicles demonstrates a gradual and significant increase. In contrast, the numbers of coated vesicles and pits are variable within a very narrow range, and no pattern of increase or decrease is discernible during development. Similarly, there is no change in interendothelial cell junctions, which consist of occluding and primitive adhesive junctional types, during development. The lamina densa of the basal lamina gradually develops from discontinuous, patchy densities along the abluminal surface of the endothelial cells to a continuous and distinct layer by 21 days gestation. The presence of the proteoglycan species in the developing basal lamina was assessed with the cationic dye ruthenium red (RR), and the appearance of RR-marked proteoglycans was found to parallel the appearance of lamina densa material. found to parallel the appearance of lamina densa material. The RR sites appear discontinuously in patches; and later, the RR sites appear in a continuous and regular planar lattice in the lamina rara interna and externa at 21 days gestation. A complete array of RR-stainable anionic sites outside a continuous lamina densa near birth indicates that the basal laminae of developing capillaries in the heart are morphologically, and in part biochemically, mature by the end of the first neonatal week. Our results show that the endothelial cells and the subtending basal lamina of myocardial capillaries gradually mature morphologically during the final days of gestation and the first neonatal week.(ABSTRACT TRUNCATED AT 400 WORDS)

Aging↗
PubMed1987

Myocardial capillaries in the fetal and the neonatal rat: a morphometric analysis of the maturing myocardial capillary bed.

Developing myocardial capillaries from 16-day-gestation fetus to adult undergo several morphological changes including a thinning of the lateral extensions of the capillary endothelial cells, the formation of a basal lamina, and an increase in the number of plasmalemmal vesicles. A decrease in the extracellular space, an increase in the number of capillaries, and a decrease in the capillary diameter were also observed during the developmental period. In view of these ultrastructural changes, a morphometric analysis was made on the developing myocardial wall to demonstrate specific quantitative changes. The volumes which were occupied by capillary endothelial cells, capillary lumina, extracellular space, and myocardial myocytes within a reference volume of myocardium were measured; and we found that 8% of the reference myocardial volume was occupied by capillary endothelial cells, 85% was occupied by myocardial myocytes, 4% was occupied by capillary lumina, and, except for a significant change in extracellular space at 16 days gestation, 3% was occupied by extracellular space. Each volume ratio was found to be nearly constant throughout the studied period. In contrast to this constancy in the volume ratios, other parameters which were measured demonstrated significant changes during the developmental period studied. These overall changes include a 135% increase in capillary density, a 63% increase in luminal surface area of capillary endothelial cells, a 24% decrease in capillary diameter, a 12% decrease in diffusion distance, and a 35% decrease in the diameter of the erythrocyte population.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Functional maturation of the capillary wall in the fetal and neonatal rat heart: permeability characteristics of developing myocardial capillaries.

The development of the functional components of the myocardial capillary wall was characterized by time-course studies of transendothelial transport of intravascularly injected probes of graded size from 16 days of gestation in the fetal rat to seven days postpartum. Despite the morphological changes occurring in the developing endothelial cells, the interaction of the probes was similar throughout the developmental period studied. The carbon particles were retained within the capillary lumina without any association with interendothelial junctions or with plasmalemmal vesicles. Carbon also was associated with coated vesicles. In contrast to carbon, ferritin was localized sequentially, over 60 sec of circulation, in plasmalemmal vesicles on the lumenal surface, in the cytoplasm, and on the ablumenal surface of the endothelial cells as well as in the interstitial space. Ferritin was located also in coated pits and vesicles and, after 90 sec of circulation, in multivesicular bodies. Within 30 sec of circulation, reaction product of myoglobin was located in plasmalemmal vesicles, coated vesicles, and transendothelial cell channels. Also within 30 sec, myoglobin partially filled the interendothelial space from the capillary lumina to the level of the tight junction. At all developmental ages studied, the interendothelial cell junctions appeared structurally tight and were impermeable to all of the probes. Once ferritin or myoglobin had reached the ablumenal space, the basal lamina did not appear to restrain the passage of the probes. Plasmalemmal vesicles are the capillary structures which transendothelially transport ferritin and myoglobin in developing myocardial capillaries.

Animals↗
PubMed1985

In-vivo endocytosis by bristle-coated pits and intracellular transport of endogenous albumin in the endothelium of the sinuses of liver and bone marrow.

The localization of endogenous albumin in the endothelia of the sinuses of bone marrow and liver of rats was studied by means of rabbit anti-rat albumin Fab fragment conjugated with horseradish peroxidase (HRP) visualized by 3,3' diaminobenzidine tetrachloride (DAB)-osmic acid. Albumin reaction product was present in the bristle-coated pits, bristle-coated vesicles, transfer tubules, and in larger vesicles of the endosome-lysosome system. The presence of endogenous albumin in the components of the vacuolar apparatus suggests that these endothelia may be active in the catabolism of endogenous albumin. Because of the ability of albumin to bind to a large variety of chemically different substances, the possible role of endogenous albumin in the internalization of certain substances by the bristle coated pits and bristle coated vesicles of these sinus endothelia needs to be considered.

Albumins↗
PubMed1983

Effects of temperature on anion distribution in perfused rat, guinea-pig and hamster ventricle.

Ventricular tissue from rats, guinea-pigs and hamsters were found to be more anion permeable when perfused and superfused with Ringer's solution at 22 degrees C rather than 36 degrees C. When the perfusate temperature was 36 degrees C the anion permeability of the in vitro rat and guinea-pig ventricle approximated that found in situ. Further, the anion permeability of the rat and guinea-pig heart was found not to be influenced by the absence of plasma proteins in the perfusate.

Animals↗
PubMed1983

Phagosome-lysosome interactions related to erythrophagocytosis in Kupffer cells of fetal rat liver.

Kupffer cells of fetal rat liver were examined by ultrastructural cytochemical methods to reveal acid phosphatase (AcPase) activity in lysosomes. Elongated cisternae, 940-1150 A in width containing AcPase reaction product, were identified in these cells. These cisternae were sometimes in continuity with phagosomes containing engulfed erythrocytes. Observations suggest that such cisternae may partly encircle these phagosomes. The relationships of these cisternae to GERL (Golgi Endoplasmic Reticulum Lysosomes) is discussed.

Acid Phosphatase↗
PubMed1983

A survey of the binding of polycationic ferritin in several fenestrated capillary beds: indication of heterogeneity in the luminal glycocalyx of fenestral diaphragms.

Polycationic ferritin (PCF) was injected into umbilical veins of 17- and 21-day-gestation rats and saphenous veins of young, male, adult Sprague-Dawley rats and allowed to circulate for 1 min or less followed by excision of tissues and immersion fixation. Fetal liver, intestine, and kidney, as well as adult liver, intestine, pancreas, kidney, adrenal cortex, bone marrow, and pituitary were examined. Nearly all fenestral diaphragms, but no subendothelial structures, were labeled with a tuft of particles in fetal intestine, adult intestine, pancreas, pituitary, and renal peritubular capillaries. A fraction of the diaphragms covering fenestrae in the thyroid, adrenal cortex, and fetal kidney glomerulus, but none in the bone marrow and fetal liver, had associated PCF. In these tissues some diaphragms appeared to be permeable to PCF, because tufts were observed immediately beneath unlabeled fenestral diaphragms either in the basal lamina (fetal kidney, adult thyroid, and adrenal cortex) or on other subendothelial structures (fetal liver, adult bone marrow). Also periodic concentrations of PCF were observed in the lamina rara interna of the adult glomerular basement membrane. PCF binding to the fenestral diaphragms and the basal lamina in the adult intestine and the renal glomerulus, respectively, have been reported by Simionescu et al. ((1981a) J. Cell. Biol. 90, 605-613) and Kanwar and Farquhar ((1979) J. Cell. Biol. 81, 137-153) as indicating accumulations of anionic material. In this study we have demonstrated not only high-specificity binding of PCF to fenestral diaphragms in capillaries of organs other than intestine and pancreas, but also considerable variation of PCF stainability of fenestral diaphragms in other organs.

Animals↗
PubMed1982

Fetal and neonatal rat intestinal capillaries: permeability to carbon, ferritin, hemoglobin, and myoglobin.

The function of the mural components involved in vascular exchange (pinocytotic vesicles, fenestrations, intercellular junctions, and the basal lamina) was assessed by intravascular injection of tracers (carbon, carbon plus histamine, ferritin, hemoglobin, and myoglobin) on separate gestational days in fetal and neonatal rat intestinal capillaries. The continuous capillaries present through day 17 of gestation were impermeable, even at junctions, to all the tracers within the maximum circulation time studied (3 minutes). Most of the luminal pinocytotic vesicles were labeled within 3 minutes with myoglobin (3.3-nm diameter) or hemoglobin (5.5-nm diameter), while only occasional vesicles contained ferritin (11-nm diameter) and none contained carbon (25-50nm diameter). Even though luminal pinocytotic vesicles contained the smaller tracers, no vesicular transport and discharge were ever seen. The fenestrated vessels present from day 18 of gestation allowed a rapid extravasation of ferritin and hemoglobin via diaphragmed fenestrations. At no time period did extravasation of carbon take place, nor did the carbon plus histamine solution induce vascular leakage. Once a tracer gained access to the extravascular space, the developing basal lamina did not seem to retard its movement.

Animals↗
PubMed1981

Fetal and neonatal rat intestinal capillaries: a TEM study of changes in the mural structure.

The development of intestinal mucosal capillaries was studied in Wistar rats from the fourteenth day of gestation through the second postnatal day with the transmission electron microscope. The microvessels develop first as continuous capillaries without a basal lamina and become fenestrated during later stages of gestation. In the thickened areas of the cytoplasm of endothelial cells at early stages, there is a paucity of pinocytotic vesicles; but some vesicles are observed which are larger than those in the adult. Later in gestation these larger vesicles can still be seen, but the frequency of their occurrence decreases with the maturation of the vessels. The intercellular junctions vary in length and shape, and the space between the outer leaflets of the apposed cell membranes is usually 20 nm. Surrounding the abluminal surface of the endothelium, the incipient basal lamina appears attenuated and incomplete until at least the second day of neonatal life.

Animals↗
PubMed1981

The relationships between the Golgi apparatus, GERL, and lysosomes of fetal rat liver Kupffer cells examined by ultrastructural phosphatase cytochemistry.

Kupffer cells are the sinusoidal macrophages of the liver. Using ultrastructural phosphatase cytochemical methods, we examined the relationship between the Golgi apparatus, GERL, and lysosomes of Kupffer cells in fetal rat livers identified, in part, by their ability to phagocytize intravenously injected latex spheres. Thiamine pyrophosphatase (TPPase) activity was localized to the inner Golgi saccules and some vesicles in the Golgi region but not to GERL. A TPPase-like activity, demonstrable in lysosomes, was abolished by sodium fluoride but not suppressed by the alkaline phosphatase inhibitors L-cysteine and L-p-bromotetramisole. Acid phosphatase (AcPase) was localized by GERL, some coated vesicles, and in lysosomes, but not to the Golgi stacks. Continuities between GERL and lysosomes were observed. Phagosomes containing internalized latex spheres received TPPase and AcPase sequentially. TPPase was localized in phagosomes immediately after latex administration. AcPase activity was not found here until at least 10 minutes following the injection of the particulates. Our findings indicate that Kupffer cell lysosomes are derived from GERL, but also suggest that phagosomes may receive material packaged by the Golgi apparatus as well as GERL.

Acid Phosphatase↗
PubMed1980

The development of the sinusoids of fetal rat liver: morphology of endothelial cells, Kupffer cells, and the transmural migration of blood cells into the sinusoids.

The fine structural development of rat fetal liver sinusoids from 10 to 22 days gestation was studied. Colloidal carbon (Pelikan ink) was injected into 14-22 day gestation fetuses via the umbilical vein to assess the continuity of the sinusoidal lining and the phagocytic ability of the developing lining cells. Endothelial cells, devoid of an underlying basal lamina, form the bulk of the vascular lining at all gestational ages. These cells possess typical intercellular junctions and fenestrae with diaphragms before 17 days gestation. Transendothelial open fenestrations, typical of the adult liver, appear around 17 days gestation, increasing in number for the remainder of gestation. Although fenestrae possessing diaphragms are permeable to carbon before 16 days gestation, open fenestrations, first seen at 17 days gestation, allowed large amounts of carbon to reach the extravascular space. Endocytosis of carbon by endothelial cells was accomplished exclusively by large bristle-coated vesicles. Endothelial cells were also seen to be involved in transmural diapedesis of newly formed erythrocytes and megakaryocyte processes from the extravascular space by forming a temporary migration pore allowing these cells and processes to enter the circulation. At the end of gestation, blood-forming activity had nearly ceased, and only the space of Dissé separated the lining cells from the parenchymal cells. Kupffer cells were easily identified as early as 13 days gestation by their content of phagosomes and engulfed erythrocytes. The Kupffer cells are much more avid in the phagocytosis of carbon than are endothelial cells. Toward the end of gestation, some Kupffer cells develop a homogeneous "sticky coat" to carbon.

Animals↗
PubMed1979

The development of the sinusoids of fetal rat liver: localization of endogenous peroxidase in fetal Kupffer cells.

Endogenous peroxidase is the cytochemical marker used to identify Kupffer cells in the adult liver. In this study, we show by ultrastructural cytochemistry that Kupffer cells of the fetal rat liver are endogenous peroxidase positive. The reaction product is localized in the endoplasmic reticulum including the perinuclear cisternae and in a few lysosome-like dense bodies. Serial sections of Golgi regions suggest that GERL and not the Golgi stacks, is peroxidase positive. As in the adult liver, peroxidase is not localized in endothelial cells. Kupffer cells do not appear to transform from endothelial or extravascular developing monocytic cells and are present prior to bone marrow formation. The relevance of these observations with respect to the possible origin of the Kupffer cell is discussed.

Animals↗