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PubMed2002

Impeller inner diameter in a miniaturized centrifugal blood pump.

To design a miniaturized centrifugal blood pump, the impeller internal diameter (ID), which is a circle diameter on the inner edge of the vane, is considered one of the important aspects. Hydraulic performance, hemolysis, and thrombogenicity were evaluated with different impeller IDs. Two impellers were fabricated with an outer diameter of 35 mm, of which 1 had an 8 mm ID impeller and the other had a 12 mm ID. These impellers were combined with 2 different housings in which the inlet port was eccentrically positioned 3.8 and 4.5 mm offset from the center. The hydraulic performance and hemolysis were evaluated in a mock circuit, and thrombogenicity was evaluated in a 2 day ex vivo study with each impeller housing combination. Both impellers required 3,000 rpm in the 3.8 mm offset inlet to attain 5 L/min against 100 mm Hg (left ventricular assist device condition). The 8 mm ID impeller required 3,200 rpm, and the 12 mm ID impeller required 3,100 rpm in the 4.5 mm offset housing. The normalized index of hemolysis was 0.0080 +/- 0.0048 g/100 L in the 8 mm ID impeller with the 3.8 mm offset and 0.022 +/- 0.018 g/100 L with 4.5 mm offset. The 12 mm ID impeller had 0.068 +/- 0.028 g/100 L with the 3.8 mm offset and 0.010 +/- 0.002 g/100 L with the 4.5 mm offset. After the 2 day ex vivo study, no blood clot was formed around the top bearing in all the pump heads. The 8 mm ID impeller with 3.8 mm offset inlet and the 12 mm ID impeller with the 4.5 mm offset had less hemolysis compared to the other pump heads that were subjected to 14 day ex vivo and 10 day in vivo studies. The 8 mm ID impeller with the 3.8 mm offset inlet had a blood clot around the top bearing after the 14 day ex vivo study. No thrombus was found around the top bearing of the 12 mm ID impeller with the 4.5 mm offset in the 10 day in vivo study. These results suggest that the ID does not greatly change the hydraulic performance of a small centrifugal blood pump. The proper combination of the impeller ID and inlet port offset obtains less hemolysis. The larger impeller ID is considered to have less thrombogenicity around the top bearing.

Centrifugation↗
PubMed2003

The role of regulatory cells in miniature swine rendered tolerant to cardiac allografts by donor kidney cotransplantation.

To determine the mechanism by which cotransplantation of a kidney allograft induces tolerance to a donor heart in miniature swine, we examined the role of CD25+ cells in heart/kidney recipients. Tolerance was induced to class-I MHC mismatched hearts by cotransplanting a donor-specific kidney with a 12-day course of cyclosporine. Peripheral blood leukocytes (PBL) were isolated from tolerant heart/kidney recipients and used in cell-mediated lympholysis (CML) coculture assays as either unmodified PBL, PBL enriched for CD25+ cells or PBL depleted of CD25+ cells to assess their ability to suppress CML responses of naïve recipient-matched leukocytes against mismatched target cells. Primed PBL from tolerant heart/kidney recipients completely suppressed lysis by naïve cells. Complete suppression of the response of naïve recipient-matched leukocytes against donor-matched target cells was lost following the depletion of CD25+ cells from tolerant heart/kidney animal PBL, but it was reestablished by incubation of naïve cells with small populations of CD25+ cells from tolerant heart/kidney animals. These data suggest that peripheral blood from tolerant heart/kidney recipients contains regulatory cells that, upon priming, can suppress the response of naïve-matched PBL in coculture CML assays, and that suppression appears to be dependent on cells expressing CD25.

Animals↗
PubMed1995

Fibre type differentiation during postnatal development of miniature pig skeletal muscles.

Histochemical differentiation of 12 skeletal muscles with a different fibre type composition was studied in miniature pigs from 80 d of gestation to 1 year of age. Two fetal myofibre types were distinguished at 100 d of gestation by the mATPase reaction after acid preincubation. The staining for oxidative enzyme activities showed no conspicuous differences between fibres up to the 6th day after birth. Starting from this age it was possible to distinguish 3 fibre categories: SO (slow-twitch oxidative); FOG (fast-twitch oxidative-glycolytic); and FG (fast-twitch glycolytic). A characteristic cluster distribution of the 3 fibre types was observed in all studied muscles with the exception of the masseter muscle which consisted only of the type SO and FOG fibres with a mosaic arrangement. The frequencies of both SO and FG fibre types increased and the proportion of type FOG fibres decreased during the postnatal period. These changes could be explained by developmental transformations among the individual fibre types. The type FOG fibres converted preferably to the fibre type (SO or FG) that prevailed in the muscles of adult animals.

Adenosine Triphosphatases↗
PubMed2000

Effects of exercise on plasma high-density lipoprotein cholesteryl ester metabolism in male and female miniature swine.

We studied the effects of exercise on high-density lipoprotein (HDL) cholesteryl ester (CE) metabolism in 6 male and 6 female miniature pigs fed a commercial swine diet supplemented with cholesterol and fat. The diets were fed for a total period of 20 weeks. During the last 12 weeks of the feeding period, the pigs were exercised on a motorized treadmill 5 days per week for 45 min/d at a speed of 9.5 to 10.0 km/h at 0% grade. Homologous HDL preparations were radiolabeled with cholesteryl (1-14C)oleate and intravenously administered to the pigs, followed by blood sampling at the appropriate time points and measurement of radiolabeled HDL CE. This was performed while the animals were sedentary and after the exercise period. Plasma cholesterol increased after the exercise protocol from 7.21 +/- 1.90 to 8.50 +/- 2.81 mmol/L (mean +/- SD, n = 6) in the females and from 8.11 +/- 3.61 to 10.07 +/- 3.61 in the males. HDL CE transport rates in female pigs were significantly lower (23%) after the exercise protocol (118 +/- 14 v 91 +/- 14 micromol/h/L plasma). HDL CE transport rates in the males were also lower (11%) after exercise (90 +/- 20 v 80 +/- 18 micromol/h/L plasma), but this effect was not statistically significant. Further, the residence time or life span of HDL CE was significantly longer after the exercise protocol in both male and female pigs. Thus, the results of this study suggest that exercise reduces the transport rate of HDL CE and prolongs the life span of HDL CE in hypercholesterolemic pigs.

Animals↗
PubMed1977

A new miniaturized system for monitoring the epidural pressure in children and adults.

A new miniaturized epidural pressure transducing system for monitoring intracranial pressure (ICP) has been developed. The system is of the as yet smallest known dimensions. The transducer is implanted automatically coplanar to the dura via a burr hole of 4 mm diameter in the conscious patient without special surgical equipment. Application is possible in adults, children and infants. We have achieved a maximal resolution of 10(-3) mm of Hg. At a full scale range of 100 mm of Hg the zero-point drift is less than 1 mm of Hg/24 h.

Adult↗
PubMed1983

Localization of prostaglandin D2 binding protein and NADP-linked 15-hydroxyprostaglandin D2 dehydrogenase in the Purkinje cells of miniature pig cerebellum.

High-affinity binding sites specific for prostaglandin D2 were visualized by the autoradiographical technique assisted by computerized image processing and color coding. In the cerebellum of miniature pig, high density of the binding was observed in the cortex, mainly in the Purkinje cell layer. Immunohistochemical studies on the cellular localization of NADP-linked 15-hydroxyprostaglandin D2 dehydrogenase were performed with the same cerebellum and revealed the presence of this enzyme also in the Purkinje cells. These findings imply that prostaglandin D2 binds to the Purkinje cells and, after its action, is degraded in situ by the specific degradative enzyme.

Animals↗
PubMed1990

Structure of miniature swine class II DRB genes: conservation of hypervariable amino acid residues between distantly related mammalian species.

As part of our studies of the class II genes of miniature swine, we have isolated and characterized cDNA clones corresponding to DRB genes from two major histocompatibility complex homozygous strains. Comparison of the sequences of these clones to those of human DRB genes revealed a striking amino acid homology between the hypervariable residues of SLA-DRBc and the human DRB1-0101 allele. The percentage of differences in these residues between the pig DRBc allele and the human DRB1-0101 allele was significantly lower (29%) than that between the DRB1-0101 allele and all other human alleles (average, 66.2%). This similarity was not seen in a comparison of the number of silent substitutions, by which the swine DRBc and the human DRB-0101 differed. Since phenotypic selection operates at the level of protein products rather than nucleotide sequences, these data suggest the existence of selective mechanisms that have resulted in similar hypervariable regions in certain alleles even in these widely disparate species. Consistent with this hypothesis, an examination of available murine and bovine class II sequences revealed a homology in hypervariable residues between the human DRB1-1401 allele and the mouse E beta s allele as well as a cow DRB allele. Consideration of these data along with intraspecies allelic sequence comparisons suggests that at least some of the interspecies similarities have emerged as the result of convergent evolution, possibly as the result of a need to react to common pathogens.

Amino Acid Sequence↗
PubMed1996

Long-term effects on lipid metabolism in miniature swine (Sus scrofa) of diets enriched in saturated, monounsaturated and polyunsaturated (N-6 and N-3) fatty acids.

In this paper we study the effect of long-term adaptation (twelve months) of lipidic parameters of miniature swine to diets enriched in saturated (lard; L-group), monounsaturated (olive-oil; O-group) and polyunsaturated (sunflower and fish-oil; S- and F-groups respectively). The experimental group with the highest level of total cholesterol, free cholesterol and phospholipids was the S-group. This group had as well levels of HDL-C and LDL-C significantly higher when compared with the remaining groups. The L-group had the lowest value of HDL-C. In spite of that, the index of artherogenicity (HDL/LDL+VLDL) was significantly higher in the L-group, followed by the O-, F- and S-group respectively. On the other hand, after 12 months adaptation we observed that the fatty acid composition of serum lipids clearly reflects the quality of the dietary fats. The O-group had significantly higher serum oleic acid levels than all the other groups, and its content in saturated fatty acids was the lowest. The same happens with red blood cell (Rbcs) membranes fatty acids but the effect is less marked. Membranes of the L-group were found to have the highest saturation index (SI) in Rbcs membranes, while the F-group had the highest unsaturation index (UI), followed by the O-group. Taken together, our findings show that the diet enriched in olive oil produces a lipid pattern intermediate between that obtained with the fish-oil-diet and the sunflower-oil-diet but with the advantage of Rbcs membranes with a lower amount of PUFAS. As it is known, membranes with high percentages of PUFAS are more accessible for peroxidation, and the degree of peroxidation of lipids is directly related with changes in the membranes functionality.

Adaptation, Physiological↗
PubMed2003

Essential role of magnesium ion in water for colonization of Helicobacter pylori in 2-week-old miniature pigs.

This study was designed to determine whether magnesium ion in water would influence the colonization of Helicobacter pylori in 2-week-old miniature pigs. Groups A (2 pigs) and B (1 pig) were both fed a milk diet dissolved in drinking water, Group C (2 pigs) was fed a milk diet dissolved in deionized distilled water (DDW), and Group D (1 pig) was fed a milk diet dissolved in DDW supplemented with MgCl2. Groups B, C, and D were all challenged with H. pylori, and Group A was not. Necropsy was performed on the pigs on postinfection Day 5, and biopsy specimens were taken from 16 sites of the stomach. H. pylori were recovered from 11 of 16 sites in Group B, 1 of 32 sites in Group C, and 13 of 16 sites in Group D. On the other hand, the degree of lymphocyte infiltration increased in the order of Group A < Group B < Group C < Group D. These observations suggest that magnesium ion in drinking water is essential for the colonization of H. pylori in the pig stomach. Possible mechanisms for the lymphocyte infiltration are discussed.

Animals↗
PubMed2002

Intralaryngeal application of a miniaturized ultrasonic probe.

We developed a method for performing intralaryngeal ultrasonography. Normal larynges were obtained from 10 cadavers and examined using an intraluminal ultrasonic tomography apparatus connected to a radial scanning 20 MHz miniaturized probe. The larynx was placed in a bath filled with physiologic saline. and the probe was inserted through the forceps channel of the fiberscope (6 mm diameter). A horizontal ultrasonic image of the vocal fold was obtained. Histologic sections of the larynx were compared with the ultrasonic images. The mucosa in the membranous region of the vocal fold was comprised of three layers ultrasonographically. The epithelium and superficial layer of the lamina propria were visualized as a high echo (hyperechoic) region, the intermediate layer of the lamina propria was visualized as a low echo (hypoechoic) region and the deep layer of the lamina propria was seen as a hyperechoic region. The vocal fold structure can be visualized by intralaryngeal ultrasonography using the filling method. This method may be clinically useful for the detection of tumors involving the vocal folds.

Animals↗
PubMedDate not supplied

A miniature suture tensiometer for laparoscopic applications.

The tension applied to sutures can affect their performance. In laparoscopic procedures, the ability to tension sutures may be compromised by the confined space. The ability to accurately judge tension may also be compromised by lever arms of long instruments and interaction between the instrument and trocar. A miniature suture tensiometer that can be used in open and laparoscopic settings has been developed that demonstrates a maximum error of 0.05 lb over a 2-lb range. This device has been used in laparoscopic surgery to measure suture tension. The suture tension information gathered has been used in the development of a device that reliably delivers a secure knot in a laparoscopic setting.

Animals↗
PubMed1984

Effect of class II antigen matching on renal allograft survival in miniature swine.

The benefit of class II major histocompatibility complex (MHC) antigen matching to renal allograft survival, in the absence of immunosuppression, has been studied in partially inbred miniature swine. Permanent (greater than 6 mo) renal allograft survival was found in 30% of recipients of either class II only or fully matched grafts. Analysis of the survival of the class II-only matched grafts by specific recipient/donor haplotype combinations indicated that survival was regulated by at least three genetic factors, including antigen gene dose, a class I MHC allele-dependent effect, and non-MHC-linked immune response phenomenon. Animals accepting class II-matched kidneys developed spontaneous tolerance to the graft, despite mounting an initial immune response marked by renal damage and the development of serum cytotoxic antibodies directed at the donor MHC antigens. The antibodies were only of the IgM class, suggesting that conversion of the humoral response to IgG was blocked. After acceptance of the kidney, three out of five animals showed specific prolongation of donor skin grafts. At the time of rejection of these skin grafts, no decrease in renal function nor reappearance of anti-donor antibodies was observed.

Animals↗
PubMed1999

Clinical relative biological effectiveness of low-energy x-rays emitted by miniature x-ray devices.

Several groups are developing ultra-miniature x-ray machines for clinical use in radiation therapy. Current systems are for interstitial radiosurgery and for intravascular insertion for irradiation to prevent re-stenosis. Typical generating voltages are low, in the 20 to 40 kV range. It is well established that the biological effectiveness of such low-energy photons is large compared with higher-energy gamma rays, because of the dominance of photoelectric absorption at low energies. We have used microdosimetric analyses to estimate RBEs for such devices, both at low doses and clinically relevant doses, relative to radiations from 60Co, 192Ir, 125I and 90Sr/90Y. The RBEs at clinically relevant doses and dose rates for these low-energy x-ray sources are considerably above unity, both relative to 60Co and to 192Ir photons, and also relative to 125I and 90Sr/90Y brachytherapy sources. As a function of depth, the overall effect of the change in dose and the change in beam spectrum results in beams whose biologically weighted dose (dose x RBE) decreases with depth somewhat more slowly than does the physical dose. The estimated clinically relevant RBEs are sufficiently large that they should be taken into account during the treatment design stage.

Brachytherapy↗
PubMed2003

Miniaturization of cell-based beta-lactamase-dependent FRET assays to ultra-high throughput formats to identify agonists of human liver X receptors.

Activation of liver X receptors (LXRs) induces reverse cholesterol transport and increases high-density lipoprotein cholesterol in vivo. Here, we describe novel, functional, homogeneous cell-based fluorescence resonance energy transfer assays for identifying agonists of LXRs using beta-lactamase as the reporter gene. Stable Chinese hamster ovary cell lines expressing LXRalpha-GAL4 or LXRbeta-GAL4 fusion proteins that regulate beta-lactamase transcription from upstream 7 x UAS GAL4 DNA binding sequences were generated and characterized. Synthetic and natural ligands of LXR dose-dependently activated the expression of beta-lactamase in a subtype-specific manner. These assays were used to demonstrate that a 1-pyridyl hydantoin small molecule LXR synthetic ligand specifically activates LXRalpha receptors. The beta-lactamase assays were optimized for cell density, dimethyl sulfoxide sensitivity, and time of agonist stimulation. Clonal LXRbeta-GAL4-beta-lactamase cells were miniaturized into an ultra high throughput (3456-well nanoplates) screening format.

Animals↗
PubMed1991

Histocompatible miniature pig (d/d haplotype): generation of hybridomas secreting A or M monoclonal antibody.

Two Hypoxanthine/Aminopterin/Thymidine-sensitive cell sublines (L142 and L231) have been derived from independent lymphoblastoid cell lines of B lineage. After propagation for more than 100 population doublings (1 year) in culture, these cells still retained a doubling time between 19 to 20 hours, near diploïdy and relatively low (L142) and high (L231) secretion rate of M immunoglobulins. Near diploid hybrid cells were easily generated with leukocytes from the spleen, the gut lamina propria or the mesenteric lymph nodes of pigs immunized against the transmissible gastroenteritis virus. Both the tumor sublines and the B cells were derived from histocompatible miniature pigs (d/d haplotype). Demonstration of fusion between the tumor sublines and B-cells was supported by the selection of hybridomas making the antigen-specific heavy (alpha isotype) and light chains from the B cell parent as well as the heavy and light chains of the lymphoblastoid parent. Moreover, some hybridomas were found to secrete only class A (dimeric) or class M immunoglobulins (0.2-10 micrograms/ml). Forty hybridomas secreted antibodies reactive in a virus-enzyme-linked cell immunoassay against cell-bound antigens and two were found to produce an antibody active only against the infected cell monolayer. Construction of intraspecies hybridoma can be used to perpetuate lymphocyte subsets useful for the study of the porcine immune system.

Animals↗
PubMed1992

Reduced tissue arachidonic acid concentration with chronic ethanol feeding in miniature pigs.

The effect of ethanol feeding on the essential fatty acid content of tissues has been contradictory. To define the effect, we analyzed fatty acid profiles in various tissues from five miniature pigs fed daily 105 kJ basal diet/kg body wt and 146 kJ ethanol/kg body wt, and also five control pigs pair-fed the same amount of basal diet but with corn starch substituted for ethanol. After 12 mo, biopsy samples were taken, and tissue fatty acid profiles were analyzed. In the phospholipid fraction from the ethanol group there was a uniform decrease in arachidonic acid (AA) and an increase in oleic acid in liver, serum, and muscle. AA was consistently decreased in the triglyceride fractions of liver, serum and subcutaneous adipose of the ethanol group. Possible explanations for this general reduction in tissue AA with ethanol feeding include decreased activities of delta 6 and delta 5 desaturases, and a displacement of AA from lipid fractions by other fatty acids.

Adipose Tissue↗
PubMed1990

Renal denervation decreases blood pressure in DOCA-treated miniature swine with established hypertension.

The role of renal sympathetic nerve activity (RSNA) in the maintenance phase of essential hypertension has not yet been clearly defined. Renal function and mean arterial pressure (MAP) were studied in four Yucatan miniature swine (YMS) with established DOCA hypertension prior to and for 3 weeks after surgical renal denervation (RDX). During the first week post-RDX, MAP decreased from 141 /+- 6 to 121 +/- 3 mm Hg (P less than .05), while sodium balance increased from 0.32 +/- 0.05 to 0.95 +/- 0.14 mEq/kg/day (P less than .05). By 3 weeks post-RDX, MAP remained below normotensive levels while sodium balance returned to the pre-RDX value. There was no significant change in potassium or water balance after RDX. Thus, in DOCA-YMS the renal nerves are important in the maintenance of hypertension. The reduction in MAP with RDX in the absence of a natriuresis suggests a role for renal afferent nerve activity.

Analysis of Variance↗
PubMed2005

Economy, speed and size matter: evolutionary forces driving nuclear genome miniaturization and expansion.

BACKGROUND: Nuclear genome size varies 300 000-fold, whereas transcriptome size varies merely 17-fold. In the largest genomes nearly all DNA is non-genic secondary DNA, mostly intergenic but also within introns. There is now compelling evidence that secondary DNA is functional, i.e. positively selected by organismal selection, not the purely neutral or 'selfish' outcome of mutation pressure. The skeletal DNA theory argued that nuclear volumes are genetically determined primarily by nuclear DNA amounts, modulated somewhat by genes affecting the degree of DNA packing or unfolding; the huge spread of nuclear genome sizes is the necessary consequence of the origin of the nuclear envelope and the nucleation of its assembly by DNA, plus the adaptively significant 300 000-fold range of cell volumes and selection for balanced growth by optimizing karyoplasmic volume ratios (essentially invariant with cell volume in growing/multiplying cells). This simple explanation of the C-value paradox is refined here in the light of new insights into the nature of heterochromatin and the nuclear lamina, the genetic control of cell volume, and large-scale eukaryote phylogeny, placing special emphasis on protist test cases of the basic principles of nuclear genome size evolution. GENOME MINIATURIZATION: and Expansion Intracellular parasites (e.g. Plasmodium, microsporidia) dwarfed their genomes by gene loss and eliminating virtually all secondary DNA. The primary driving forces for genome reduction are metabolic and spatial economy and cell multiplication speed. Most extreme nuclear shrinkage yielded genomes as tiny as 0.38 Mb (making the nuclear genome size range effectively 1.8 million-fold!) in some minute enslaved nuclei (nucleomorphs) of cryptomonads and chlorarachneans, chimaeric cells that also retain a separate normal large nucleus. The latter shows typical correlation between genome size and cell volume, but nucleomorphs do not despite co-existing in the same cell for >500 My. Thus mutation pressure does not inexorably increase genome size; selection can eliminate essentially all non-coding DNA if need be. Nucleomorphs and microsporidia even reduced gene size. Expansion of secondary DNA in the main nucleus, and in large-celled eukaryotes generally, must be positively selected for function. Ciliate nuclear dimorphism provides a key test that refutes the selfish DNA and strongly supports the skeletal DNA/karyoplasmic ratio interpretation of genome size evolution. GENETIC CONTROL OF CELL VOLUME IS MULTIGENIC: The quantitatively proportional correlation between genome size and cell size cannot be explained by purely mutational theories, as eukaryote cell volumes are causally determined by cell cycle control genes, not by DNA amounts.

Animals↗