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PubMed2004

Pregnancy diagnosis in miniature pig by direct ELISA of oestrone derivatives in faeces.

The objectives of the present study were to measure oestrone derivatives [oestrone, oestrone sulphate (E1S) and oestrone glucuronide] in sow faeces by direct enzyme-linked immunosorbent assay (ELISA) and to explore the changes of oestrone derivative profile in faeces of miniature pig for demonstrating the possibility of pregnancy diagnosis. Faecal samples (1.5 g) were put into 6 ml of buffer, shaken and centrifuged. Then supernatant was added to the wells of multi-plate without extraction and incubated with anti-oestrone antibody and horseradish peroxidase-labelled oestrone. Standard solution was prepared at various concentrations of E1S. Sensitivity was estimated as 0.035 ng/ml (0.14 ng/g). Intra- and inter-assay coefficient variations were 3.5-7.7% and 10.9-15.3%, respectively. When 1-5 ng/ml E1S were added to a faecal solution, recovery rates ranged between 80.0 and 103.3%. There is a temporal increase in the E1S equivalent concentration of miniature pig faeces from day 25 to 31 after mating. From day 35 to 70, the E1S equivalent concentrations remained low. Thereafter its concentrations increased again towards farrowing. On day 27 and 29 after mating pregnancy diagnosis by the faecal E1S equivalent concentrations agreed with the results of farrowing (seven of seven animals). These results suggest that the present direct ELISA is practical and suitable as a routine assay for measuring the faecal concentration of oestrone derivatives and that this assay might be usable for pregnancy diagnosis in sows at day 27-29.

Animals↗

PubMed2000

Development of a miniature undulation pump for the distributed artificial heart.

Research of the distributed artificial heart is important not only to acquire the means of individual organ perfusion but also to clarify the characteristics of the organ and the mechanism of blood distribution. To investigate the distributed artificial heart, the miniature undulation pump was developed. The outer diameter and the thickness of the developed pump were 38 mm and 11 mm, respectively. The priming volume of the pump was 3.2 ml. The total size including the motor unit was 38 mm in diameter and 32 mm in length. The total weight was 67.5 g. The total volume was 27.5 ml. The pump was driven with pulse width modulation by using a 1 chip motor controller. More than 5 L/min of continuous output could be obtained. The results showed that the developed miniature undulation pump system had enough performance for individual organ perfusion.

Biomechanical Phenomena↗

PubMed2004

Effect of acetazolamide on the optic disc oxygenation in miniature pigs.

BACKGROUND: The purpose of our study was to evaluate the variations of the optic disc PO (2) during normoxia and hyperoxia (100 % O (2)), before and after intravenous administration of acetazolamide. MATERIAL AND METHODS: PO (2) measurements were obtained at intervascular areas of the optic disc in 11 anaesthetized miniature pigs using oxygen-sensitive microelectrodes introduced through the vitreous cavity by a micromanipulator. PO (2) was measured continuously during 10 minutes under systemic normoxia and systemic hyperoxia. Oxygen measurements were repeated under these conditions after intravenous injection of acetazolamide (bolus of 500 mg) in 8 animals. RESULTS: In systemic hyperoxia, the optic disc PO (2) increased moderately (DeltaPO (2) = 4.7 +/- 2.5 mmHg; p < 0.001; n = 11) in parallel with systemic PaO (2). Acetazolamide led to a slow and progressive increase in the optic disc PO (2) (DeltaPO (2) = 2.1 +/- 1.7 mmHg; p > 0.1; n = 8 after 10 min, while DeltaPO (2) = 4.3 +/- 3.2 mmHg; p < 0.05; n = 8 after 30 min), in parallel with a slow and progressive increase in systemic PaCO (2). The optic disc PO (2) increased much more significantly after injection of acetazolamide under systemic hyperoxia (DeltaPO (2) = 13.3 +/- 3.1 mmHg; p < 0.001; n = 8). CONCLUSIONS: Systemic hyperoxia alone is not sufficient to increase substantially the optic disc PO (2) in miniature pigs due to a vasoconstrictor effect. Intravenous injection of acetazolamide can increase the optic disc PO (2) progressively, due to a vasodilatory effect of elevated systemic PaCO (2). The association of acetazolamide injection with systemic hyperoxia can further improve the oxygenation of the optic disc.

Acetazolamide↗

PubMed2004

Effect of visual stimulation on blood oxygenation in the optic nerve head of miniature pigs: a pilot study.

BACKGROUND: Visual stimulation is increasingly used to investigate the coupling between neuronal activity, blood flow and metabolism in the neural tissue of the ocular fundus. In an attempt to clarify whether the oxygen metabolism is involved in this coupling, we investigated the changes in the partial pressure of oxygen of venous blood (pO (2,blood)) in the optic nerve head of pigs in response to two different visual stimuli. MATERIALS AND METHODS: In 3 miniature pigs, the pO (2,blood) was measured in the optic disk rim using the technique of phosphorescence quenching by oxygen. This parameter was recorded every 8 seconds during a dark-to-light transition and during diffuse luminance flicker (field of 30 degrees centered at the optic disk, temporal frequencies of 2 to 80 Hz). RESULTS: The venous pO (2,blood) level (mean +/- standard deviation) did not change between dark- and light-adapted conditions (26.2 +/- 5.3 and 26.0 +/- 6.2 mm Hg, respectively), nor did we observe any transient change of pO (2,blood) during the light adaptation phase. On the other hand, the venous pO (2,blood) increased, on average, relative to its level during continuous light conditions (24.5 +/- 1.9 mm Hg) by at least 6 % for all flickering frequencies, with a maximum response of 14 % at 15 Hz. CONCLUSIONS: The phosphorescence quenching technique can reveal changes in venous pO (2,blood) induced by visual stimulation. Our results show that the pO (2,blood) in the optic nerve head of miniature pigs does not change with the light adaptation state of the retina, but increases during flicker stimulation with a band-pass type response. The previously reported increase of the ONH blood flow in response to flicker stimulation could lead to this increase of pO (2,blood).

Adaptation, Ocular↗

PubMed1982

Portal vein insulin response of glucose intolerant Yucatan miniature swine to common secretogogues.

A group of glucose intolerant miniature swine exhibiting an impaired portal vein insulin response to an IVGTT were examined with respect to their portal vein insulin response to the secretogogues: isoproterenol, arginine and leucine. Equivalent insulin responses to isoproterenol and leucine were noted on the part of the glucose intolerant animals when compared to control subjects. An impaired portal vein insulin response was evident during an infusion of 0.5 g/kg arginine and again when a pulse injection of .25 g/kg glucose was administered in the presence of isoproterenol (.05 microgram/kg . min). The close agreement of these results with those reported for human diabetics suggests that a similar pancreatic defect, most probably associated with the glucoreceptor, is present in this group of glucose intolerant miniature swine.

Animals↗

PubMed2000

Miniaturized metalloproteins: application to iron-sulfur proteins.

The miniaturization process applied to rubredoxins generated a class of peptide-based metalloprotein models, named METP (miniaturized electron transfer protein). The crystal structure of Desulfovibrio vulgaris rubredoxin was selected as a template for the construction of a tetrahedral (S(gamma)-Cys)(4) iron-binding site. Analysis of the structure showed that a sphere of 17 A in diameter, centered on the metal, circumscribes two unconnected approximately C(2) symmetry related beta-hairpins, each containing the -Cys-(Aaa)(2)-Cys- sequence. These observations provided a starting point for the design of an undecapeptide, which self assembles in the presence of tetrahedrally coordinating metal ions. The METP peptide was synthesized in good yield by standard methodologies. Successful assembly of the METP peptide with Co(II), Zn(II), Fe(II/III), in the expected 2:1 stoichiometry, was proven by UV-visible and circular dichroism spectroscopies. UV-visible analysis of the metal complexes indicated the four Cys ligands tetrahedrally arrange around the metal ion, as designed. Circular dichroism measurements on both the free and metal-bound forms revealed that the metal coordination drives the peptide chain to fold into a turned conformation. NMR characterization of the Zn(II)-METP complex fully supported the structure of the designed model. These results prove that METP reproduces the main features of rubredoxin.

Circular Dichroism↗

PubMed2002

Contribution of ferrous iron to maintenance of the gastric colonization of Helicobacter pylori in miniature pigs.

Our previous study showed that the colonization levels of Helicobacter pylori were higher in the stomachs of 5-day-old miniature pigs than in 2-week-old ones. As dietary factors can cause these differences, we compared two diets, i.e., Weanymilk and a similar formula with a higher concentration of Fe(II), Weanylobulin. The colonization levels in the fundic mucosa were significantly higher in 2-week-old pigs fed Weanylobulin than in those fed Weanymilk. Supplementing Weanylobulin with an iron chelator, deferoxamine mesylate, significantly lowered the bacteria counts in the gastric mucosa. Normal diets supplemented with Fe(II) in 2-month-old pigs caused significantly more sites of bacteria in the antrum compared with normal diets alone. In addition, ranitidine, an inhibitor of gastric acid secretion that reduces Fe(III) to Fe(II) in the stomach, decreased the bacteria counts in 10-month-old pigs. These results suggested that Fe(II) maintained the colonization levels of H. pylori in the stomach of the miniature pigs.

Animals↗

PubMedDate not supplied

A miniature bidirectional RF communication system for micro gastrointestinal robots.

This paper reports a miniature, low power, two-channel, bidirectional wireless communication system that can be used in the first generation of micro gastrointestinal (GI) robots. The system consists of a miniature RF transceiver embedded in the GI robot and a control station outside the body. ISM band radio frequency (approx. 433 MHz) was used to achieve half duplex communication between the GI robot and the control station. The Frequency Shift Keying (FSK) modulation scheme was adopted to ensure a reliable and high-speed digital RF link. Animal tests have been carried out to prove the performance of the communication system.

Animals↗

PubMed1997

Role of the thymus in transplantation tolerance in miniature swine. I. Requirement of the thymus for rapid and stable induction of tolerance to class I-mismatched renal allografts.

The almost uniform failure in transplant patients of tolerance-inducing regimens that have been found to be effective in rodents, has made it necessary to examine large animal models before testing of new approaches clinically. Miniature swine have been shown to share many relevant immunologic parameters with humans, and because of their reproducible genetics, have proved extremely useful in providing such a large animal model. We have previously shown that indefinite systemic tolerance to renal allografts in miniature swine is induced in 100% of cases across a two-haplotype class I plus minor histocompatibility antigen disparity by a 12-d course of Cyclosporine A (CyA), in contrast to irreversible rejection observed uniformly without CyA treatment. In the present study, we have examined the role of the thymus during the induction of tolerance by performing a complete thymectomy 21 d before renal transplantation. This analysis demonstrated a striking difference between thymectomized and nonthymectomized animals. Thymectomized swine developed acute cellular rejection characterized by a T cell (CD25(+)) infiltrate, tubulitis, endothelialitis and glomerulitis, and anti-donor CTL reactivity in vitro. Nonthymectomized and sham thymectomized animals had a mild T cell infiltrate with few CD25(+) cells and no anti-donor CTL response in vitro. These results indicate that the thymus is required for rapid and stable induction of tolerance.

Animals↗

PubMed2004

Miniaturized, ultra-high throughput screening of tyrosine kinases using homogeneous, competitive fluorescence immunoassays.

Assay miniaturization and the implementation of high-density 1,536-microwell screening increase the speed and efficiency of screening and lead discovery. To serve this need, a platform of miniaturizable assay technologies has been assembled for specific biological targets. This platform will enable initiation and completion of uHTS screens in a straightforward and expeditious manner. Although faster primary screening does contribute to a reduction in timelines, the process of assay development can become a bottleneck. Assay technologies that do not require the use of target-specific reagents can reduce the time necessary for assay development. Assays that measure inhibition of tyrosine kinases can be configured in a competitive format where only the enzyme itself is specific to the assay. In this context, several technologies, including time-resolved fluorometry (also known as DELFIA), time-resolved fluorescence resonance energy transfer (also known as LANCE( trade mark )), fluorescence polarization, enzyme fragmentation complementation assay, and confocal laser scanning imaging, were examined. Quality parameters such as assay reproducibility, signal:background ratio, Z factor, and assay sensitivity were compared. Additionally, the relative merits of each of these technologies are assessed in terms of assay miniaturization, ease of development, ultimate screening capability, efficiency, and cost.

Amino Acid Sequence↗

PubMed2005

Piezo- and solenoid valve-based liquid dispensing for miniaturized assays.

Miniaturization of biological assays requires dispensing liquids in the submicroliter range of volumes. Accuracy and reproducibility of dispensing this range depend on both the dispenser and the receptacle in which the assay is constructed. Miniaturization technologies developed by Aurora Discovery, Inc. (San Diego, CA) include high-density multiwell plates for assay samples and reagent storage, as well as piezo-based and solenoid valve-based liquid dispensers. Some basic principles of small-volume dispensing by jetting are described to provide context for dispenser design and function. Performance of the latest instruments incorporating these dispensing devices is presented.

Biological Assay↗

PubMed2006

In vitro development of cloned embryos derived from miniature pig somatic cells after activation by ultrasound stimulation.

The present study was carried out to examine the activation and development of cloned embryos produced by transferring miniature pig somatic cells into enucleated farm pig oocytes after exposing to ultrasound. The rates of the pronucleus-like structure formation and polar body-like structure extrusion in embryos exposed to ultrasound did not differ from those applied electric pulses. Although there was no significant difference in the blastocyst formation rates between different activation methods, the mean number of cells in the blastocysts developed from embryos activated by exposing to ultrasound was significantly (p < 0.05) higher than that obtained by applying electric pulses. The results of the present study showed that ultrasound stimulation can induce the activation and in vitro development of cloned embryos derived from miniature pig somatic cells.

Animals↗

PubMed1991

Calcium and phosphorus deficiencies affect mineral distribution in neonatal miniature piglets.

Current efforts to monitor the mineral status of preterm infants fed human milk may not provide sufficient information on the distribution of body minerals. To investigate the body distribution of calcium and phosphorus during various degrees of mineral deficiency, neonatal miniature piglets were raised for 2 wk on diets differing only in calcium and phosphorus. Groups A, B, and C were fed 100%, 60%, and 20%, respectively, of the recommended amounts of calcium and phosphorus that, when adjusted for rates of growth, approximated the range of dietary intakes of preterm infants. Group C manifested biochemical and body-composition evidence of mineral deficiency when compared with group A: lower serum phosphorus; higher serum alkaline phosphatase activity; less fat-free tissue, calcium, and phosphorus in tibiae, vertebrae, and whole carcasses. Neonatal miniature piglets are useful for studying mineral deposition during mineral deficiency in preterm infants.

Alkaline Phosphatase↗

PubMed1987

Distribution and metabolism of N'-nitrosonornicotine in the miniature pig.

The distribution and metabolism of [5-3H]N'-nitrosonornicotine ([5-3H]NNN) was studied in three 18-day-old miniature pigs. [5-3H]NNN was administered by intracardiac administration into the right ventricle of the heart to mimic uptake by the lung. Whole body autoradiograms taken 15-220 min after treatment showed high levels of radioactivity in the mandibular and parotid salivary glands, Harder's gland, lacrimal glands, glands of the snout and respiratory part of the nasal cavity, and the melanin of the eyes and skin. Bound radioactivity was most abundant in the nasal mucosa and liver. Analysis of tissues by h.p.l.c. showed the presence of high levels of [5-3H]NNN in the mandibular glands and Harder's gland. Levels of [5-3H]NNN and its metabolites were determined in arterial and venous serum, 0.5-220 min after injection. The disappearance of [5-3H]NNN from serum was biphasic. 4-Oxo-4-(3-pyridyl) butyric acid, a metabolite of [5-3H]NNN resulting from 2'-hydroxylation, which is a suspected activation pathway, was detected 0.5 min after injection and appeared to reach a steady state 2-220 min after injection. 4-Hydroxy-4-(3-pyridyl)butyric acid, from 5'-hydroxylation of [5-3H]NNN, and norcotinine, from denitrosation, were also rapidly formed. These experiments are the first in which the appearance of NNN metabolites in blood has been measured. The ratio of 2'-hydroxylation to 5'-hydroxylation varied from 0.27 to 0.60 in arterial serum and from 0.33 to 0.49 in venous serum in the period from 2-220 min. [5-3H]NNN accumulated in the stomach contents such that its levels were greater than those in arterial or venous serum, 60 min after injection. The results of this study demonstrate that the miniature pig is a useful model for the investigation of nitrosamine metabolism and indicate some similarities and differences in metabolism and distribution compared with the rat.

Animals↗

PubMed1999

Laparoscopy-assisted intrapelvic sonography with a high-frequency, real-time miniature transducer for assessment of the Fallopian tube: a preliminary report.

Our purpose was to visualize normal and abnormal Fallopian tubes using laparoscopy-assisted intrapelvic sonography with a specially developed 20 MHz flexible catheter-based high-resolution, real-time miniature (2.4 mm outer diameter) ultrasound transducer in infertile women. A total of 21 women (20 infertile, one with unilateral hydrosalpinx, and one tubal pregnancy) were studied with pelvic saline effusion under laparoscopy. Fimbriae were clearly depicted with a cockscomb-like form in 95% of patients. All ampullae were visualized, and mucosal layers were clearly distinguished from muscle layers in 70% of patients. Scanty intratubal effusion was noted in 50% of patients, and tubal spastic findings were found in 10% of patients. In all, 60% of isthmuses were detected, and mucosal layers were distinguished from muscle layers in 30%. In the subject with hydrosalpinx, the tubal wall was thinner, and it was not possible to distinguish between muscle and mucosal layers. In the subject with a tubal pregnancy, the amniotic membrane and decidua were depicted more clearly than by transvaginal sonography. In conclusion, laparoscopy-assisted intrapelvic sonography with a high-frequency, real-time miniature transducer may be useful in the assessment of tubal texture and function in tubal disorders, possibly in infertility practice.

Fallopian Tube Diseases↗

PubMed1982

Influence of genetics and diet on the development of diabetes in Yucatan miniature swine.

The intent of this study was to examine the effect of a diet nutritionally similar to that consumed by most Americans on Yucatan miniature swine that have been bred for enhanced and impaired glucose tolerance. Seventeen Yucatan miniature pigs from the glucose-intolerant line (low K) and 17 from the glucose-tolerant line (high K) were selected. The oral glucose tolerance (OGT) of both groups of pigs was determined. After the OGT test (OGTT), 13 low K pigs and 12 high K pigs were placed on the American Swine Diet (ASD). The remaining animals were left on the regular swine diet. After 5 months, the pigs were again tested with an OGTT. Mean plasma glucose levels observed during the OGTT showed significant differences between the low K and high K groups on ASD at all time intervals from 60 to 150 minutes postdosing. Serum insulin levels in the high K group were significantLy elevated over those in the low K group during the OGTT at 10, 20, and 30 minutes prediet, and at 10 minutes after 5 months on diet. Both low and high K groups showed a dietary effect with aN elevated plasma glucose response to an OGTT, although the mean increase was not significant in the high K group. In the low K group, plasma glucose levels were significantly elevated (P less than 0.05) over prediet values at 20, 30, 60, 90 and 120 minutes. In addition, 7 out of the 10 low K pigs tested at 5 months developed insulin resistance, and noninsulin-dependent diabetes.

Animals↗

PubMed1994

Dietary fat saturation and endurance exercise alter lipolytic sensitivity of adipocytes isolated from Yucatan miniature swine.

This study examined the effects of dietary fat saturation and endurance exercise on lipolytic sensitivity of adipocytes isolated from Yucatan miniature swine. Twenty-four female swine had free access to a high fat diet with a polyunsaturated to saturated fat ratio (P:S) of 0.3 or 1.0, and were treadmill-exercised or remained sedentary. After 3 months, biopsies were taken, adipocytes were isolated and lipolytic activity was determined. Adipocytes were incubated with adenosine deaminase followed by epinephrine, isoproterenol, or epinephrine plus phenylisopropyladenosine, and glycerol release was measured. Backfat thickness was measured by ultrasonography. Our findings revealed that 1) adipocytes from 1.0 P:S diet-fed swine released 30% more glycerol than adipocytes from 0.3 P:S diet-fed swine when stimulated by 1 micromol/L isoproterenol; 2) adipocytes from exercised swine released 45% more glycerol than adipocytes from sedentary swine when stimulated by 1 micromol/L epinephrine; 3) body weight of exercised swine was significantly lower than sedentary swine; and 4) backfat thickness was less in exercised swine than in sedentary swine (2.39 vs. 2.95 cm, P = 0.002). We conclude that ad libitum consumption of diet with a P:S of 1.0, combined with endurance exercise, increases lipolytic sensitivity, lowers body weight gain, and reduces fat accumulation in female Yucatan miniature swine.

Adipocytes↗

PubMed1999

The sodium concentration of enteral diets does not influence absorption of nutrients but induces intestinal secretion of water in miniature pigs.

Contradictory opinions exist as to whether the sodium concentration of enteral diets influences absorption of macronutrients and transepithelial movement of sodium and water. Therefore, we investigated the effects of various sodium concentrations of enteral diets on absorption of macronutrients and on net fluxes of sodium and water. In unanesthetized miniature pigs, a 150-cm jejunal segment was perfused with an oligopeptide (Peptisorb), an oligomeric and a polymeric diet. The polymeric diet was supplemented with pancreatic enzymes. The sodium concentrations varied between 30 and 150 mmol/L. The energy density was 3.4 MJ/L. The sodium concentration of the diets did not influence absorption of macronutrients and of total energy. However, increasing sodium concentrations of the diets were associated with increasing osmolality of the solutions, resulting in a linear increase in net secretion of water and flow rate of chyme. With all diets and sodium concentrations net secretion of sodium occurred. The sodium secretion was independent of the initial sodium concentration of the diets. It was linearly correlated with net flux of water and was largest in miniature pigs infused with the oligomeric diet. The sodium concentration of the jejunal effluent did not correspond to the initial sodium concentration of the diets. The present results indicate that enteral feeding of diets with high energy density inevitably increases net secretion of water and sodium as sodium concentration increases. Therefore, the sodium concentration of diets should be as low as possible to meet only the minimal daily requirement of sodium. Low sodium concentrations of diets have no negative effects on absorption of macronutrients.

Animals↗