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PubMed2004

Absence of replication-competent human-tropic porcine endogenous retroviruses in the germ line DNA of inbred miniature Swine.

The potential transmission of porcine endogenous retroviruses (PERVs) has raised concern in the development of porcine xenotransplantation products. Our previous studies have resulted in the identification of animals within a research herd of inbred miniature swine that lack the capacity to transmit PERV to human cells in vitro. In contrast, other animals were capable of PERV transmission. The PERVs that were transmitted to human cells are recombinants between PERV-A and PERV-C in the post-VRA region of the envelope (B. A. Oldmixon, J. C. Wood, T. A. Ericsson, C. A. Wilson, M. E. White-Scharf, G. Andersson, J. L. Greenstein, H. J. Schuurman, and C. Patience, J. Virol. 76:3045-3048, 2002); these viruses we term PERV-A/C. This observation prompted us to determine whether these human-tropic replication-competent (HTRC) PERV-A/C recombinants were present in the genomic DNA of these miniature swine. Genomic DNA libraries were generated from one miniature swine that transmitted HTRC PERV as well as from one miniature swine that did not transmit HTRC PERV. HTRC PERV-A/C proviruses were not identified in the germ line DNAs of these pigs by using genomic mapping. Similarly, although PERV-A loci were identified in both libraries that possessed long env open reading frames, the Env proteins encoded by these loci were nonfunctional according to pseudotype assays. In the absence of a germ line source for HTRC PERV, further studies are warranted to assess the mechanisms by which HTRC PERV can be generated. Once identified, it may prove possible to generate animals with further reduced potential to produce HTRC PERV.

Amino Acid Sequence↗

PubMed1989

Environmental and health studies of farm workers in Swedish swine confinement buildings.

The relation between the health of workers and the environment in swine confinement buildings was investigated in a study of 57 workers on 30 swine farms in southern Sweden and 55 matched controls. Swine workers reported significantly higher frequencies of respiratory symptoms, more frequent colds and absence due to chest illness, and a history of pneumonia. The increased frequency of symptoms of respiratory disease was related to the number of years and percent of the day spent working with swine. Symptoms were also associated with respirable dust, total dust, endotoxin in total dust, and number of microbes in the air of the work environment. In a multiple regression analysis of the relation between 16 different environmental parameters to work period shifts of five pulmonary function parameters, endotoxin was found to be significantly related to the FEV1 in a dose dependent way.

Agricultural Workers' Diseases↗

PubMed2001

Head and neck hypervascular lesions: embolization with ethylene vinyl alcohol copolymer--laboratory evaluation in Swine and clinical evaluation in humans.

PURPOSE: (a) To assess in swine long-term (12-month) histopathologic changes, particularly, those related to recanalization and angiotoxicity after endovascular delivery of ethylene vinyl alcohol copolymer (EVAC), and (b) to evaluate initial clinical experience in 18 patients with head and neck tumors and arteriovenous malformations. MATERIALS AND METHODS: Embolization with EVAC was performed in one rete each in five swine. After 12 months, an angiogram was obtained, and the contralateral rete was also embolized (acute). Swine were sacrificed and the retia harvested for pathologic examination. In the clinical study, 18 patients with tumors (n = 14), facial arteriovenous malformations (n = 3), and vertebral arteriovenous fistula (n = 1) underwent therapeutic embolization. The technical aspects of EVAC embolization, percentage of occlusion, and clinical complications were evaluated. RESULTS: Angiographic 12-month follow-up in swine revealed persistent occlusion of the embolized rete or retia. Histologic examination of the same rete showed vascular occlusion and moderate intraluminal foreign body giant cell reaction; the acutely embolized rete showed no endothelial denudation or angionecrosis. Clinical evaluation in patients revealed satisfactory penetration of lesion vasculature with EVAC when the microcatheter was advanced within 2 cm of a lesion or when percutaneous puncture was performed. There were two transient complications: one increase in a preexisting fifth nerve palsy and one increase in preexisting hemiparesis. CONCLUSION: EVAC is a promising liquid embolic material providing long-term occlusion of blood vessels.

Adolescent↗

PubMed2003

Detection of active colonic hemorrhage with use of helical CT: findings in a swine model.

PURPOSE: To evaluate the feasibility of helical computed tomography (CT) as an imaging modality for depicting active colonic hemorrhage in a swine model. MATERIALS AND METHODS: Controlled extravasation of contrast material-enhanced blood (CEB) from 140 to 180 HU and at varying rates (0.3-1.0 mL/min) was performed during a 30-second period by using a microcatheter system placed within the descending colon of 14 swine. CEB was immediately followed by extravasation of unopacified blood at the same location and rate during serial helical CT imaging of the extravasation site. Region-of-interest analysis allowed quantification of the dilution of extravasated CEB as a function of time that was then modeled mathematically based on iodine mass and volume balances. Nonlinear least squares analysis was performed to optimize fitting of the model to experimental data, with a maximum regression value (R2) of 1.0 indicating a perfect fit. This model enabled the computer simulation of CT imaging of multiple combinations of bleeding rates and CEB attenuation to determine the sensitivity of helical CT for depicting active colonic bleeding. RESULTS: Sixteen swine examinations yielded 16 CEB dilution curves. An excellent fit of the model to each dilution curve was achieved, as indicated by a mean R2 value of 0.8402. Swine examinations alone showed that targeted CT could depict CEB as low as 111 HU extravasating at a rate of 0.3 mL/min. Simulations that were based on helical CT images with 5-mm collimation reconstructed every 3 mm and that used the model indicated bleeding rates below 0.4 mL/min are detectable, provided peak aortic enhancement reaches 100 HU. CONCLUSION: Conservatively, helical CT has the potential to depict active colonic hemorrhage at rates of 0.5 mL/min or less.

Animals↗

PubMed2004

MR imaging of the stomach: potential use for mangafodipir trisodium--a study in swine.

PURPOSE: To evaluate mangafodipir trisodium as a potential contrast agent at magnetic resonance (MR) imaging of the stomach. MATERIALS AND METHODS: Mangafodipir trisodium was injected intravenously into three swine at a dose of 5 micromol per kilogram of body weight. For comparison, gadopentetate dimeglumine was injected into three other swine at a dose of 0.1 mmol per kilogram of body weight. T1-weighted three-dimensional MR images were acquired in all six swine at 1.5 T before and approximately 10, 15, 20, 25, 30, and 40 minutes after contrast material administration. Extracted stomach specimens were imaged at 3.0 T. In vivo and ex vivo images were evaluated visually and quantitatively for contrast enhancement of the stomach, and in vivo images were evaluated for the presence of reflux from the duodenum. RESULTS: Mangafodipir trisodium produced prolonged and selective enhancement of the inner surface of the stomach, in contrast to the more general enhancement seen with gadopentetate dimeglumine, and reflux from the duodenum could not account for this selective enhancement. Ex vivo images confirmed that T1 enhancement in the stomach wall with mangafodipir trisodium was limited to the inner surface. Gadopentetate dimeglumine did not produce selective enhancement of the inner surface of the stomach. CONCLUSION: Mangafodipir trisodium preferentially enhances the inner surface of the stomach on MR images acquired in swine and, therefore, may have potential for use as a contrast agent at MR imaging of the human stomach.

Animals↗

PubMed2002

Regulation of apo A-IV transcription by lipid in newborn swine is associated with a promoter DNA-binding protein.

Dietary lipid acutely upregulates apolipoprotein (apo) A-IV expression by sevenfold at the pretranslational level in neonatal swine jejunum. To determine the mechanism of this regulation, two-day-old female swine received intraduodenal infusions of low- and high-triacylglycerol (TG) isocaloric diets for 24 h. Nuclear runoff assay confirmed apo A-IV gene transcriptional regulation by the high-TG diet. Footprinting analysis using the swine apo A-IV proximal promoter sequence (+14 to -246 bp) demonstrated three regions protected by the low-TG extracts. Of these three motifs, only ACCTTC showed 100% homology to the human sequence and was further studied. EMSA was performed using probes containing wild-type (WT) and mutant (M) motifs. A shift was noted with the low-TG nuclear extracts with the WT probe but not with the M probe. Excess unlabeled free WT probe competed out the shift, whereas the M probe did not. No significant shift occurred with either probe using high-TG extracts. These results suggest that a repressor protein binds to the ACCTTC motif and becomes unbound during lipid absorption, allowing transcriptional activation of the apo A-IV gene in newborn swine small intestine.

Actins↗

PubMed2003

Coronary blood flow regulation in exercising swine involves parallel rather than redundant vasodilator pathways.

In dogs, only combined blockade of vasodilator pathways [via adenosine receptors, nitric oxide synthase (NOS) and ATP-sensitive K+ (KATP) channels] results in impairment of metabolic vasodilation, which suggests a redundancy design of coronary flow regulation. Conversely, in swine and humans, blocking KATP channels, adenosine receptors, or NOS each impairs coronary blood flow (CBF) at rest and during exercise. Consequently, we hypothesized that these vasodilators act in parallel rather than in redundancy to regulate CBF in swine. Swine exercised on a treadmill (0-5 km/h), during control and after blockade of KATP channels (with glibenclamide), adenosine receptors [with 8-phenyltheophylline (8-PT)], and/or NOS [with Nomega-nitro-l-arginine (l-NNA)]. l-NNA, 8-PT, and glibenclamide each reduced myocardial O2 delivery and coronary venous O2 tension. These effects of l-NNA, 8-PT, and glibenclamide were not modified by simultaneous blockade of the other vasodilators. Combined blockade of KATP channels and adenosine receptors with or without NOS inhibition was associated with increased H+ production and impaired myocardial function. However, despite an increase in O2 extraction to >90% during administration of l-NNA + 8-PT + glibenclamide, vasodilator reserve could still be recruited during exercise. Thus in awake swine, loss of KATP channels, adenosine, or NO is not compensated for by increased participation of the other two vasodilator mechanisms. These findings suggest a parallel rather than a redundancy design of CBF regulation in the porcine circulation.

ATP-Binding Cassette Transporters↗

PubMed1981

Femoral circulatory responses to lumbar nerve stimulation in developing swine.

The maturation of femoral circulatory responses to efferent lumbar nerve stimulation was tested in 51 developing swine (1 day-3 mo old) under pentobarbital sodium anesthesia (10-30 mg/kg). Aortic pressure, heart rate, and femoral and carotid arterial flows (measured by electromagnetic flow transducers) were recorded simultaneously. Femoral vascular resistance was calculated as mean aortic pressure/mean flow. Transection of the lumbar nerve fibers below the last ganglion in the sympathetic chain did not after femoral resistance in day-old animals but decreased femoral resistance in swine 1 wk of age and older. Efferent lumbar nerve stimulation at various combinations of frequencies and intensities revealed an atropine-blockable vasodilator component in the femoral circulatory response in swine 1 mo of age and older. After alpha-adrenergic receptor blockade with phentolamine (0.25 or 0.5 mg/kg), femoral vasodilation occurred during low-frequency and -intensity stimulation of the lumbar nerve only in animals 1 mo of age and older. Acetylcholine (2 micrograms ia) caused a decrease in femoral resistance at all ages. Vasoconstrictor effects of high-frequency stimulation (5-10 Hz) were present at all ages and were age dependent. The results of these experiments suggest that the femoral circulation in swine at birth in innervated by functionally active vasoconstrictor fibers, which do not provide a tonic influence on femoral resistance until late in the first postnatal week. Furthermore, although femoral vascular cholinergic receptors are demonstrable at birth, there appears to be a delay in the maturation of functionally active vasodilator fibers.

Aging↗

PubMed1983

Acid-base curve and alignment nomograms for swine blood.

To construct acid-base curves and alignment nomograms for swine blood, we added known amounts of acid or base to aliquots of swine blood with three different concentrations of hemoglobin. Pairs of blood samples were equilibrated at two different CO2 partial pressures, and blood pH was measured. Data were analyzed by computer, and mean values were created for complete data of four swine. Curve and alignment nomograms were constructed by computer. The resultant nomograms for swine blood differ from those constructed by Siggaard-Andersen for human blood, most importantly at base-excess values of +10 to +25 mmol/l. The possible reasons for the observed differences are discussed and, although not completely resolved, may be related in large measure to the accuracy of pH determination and the methodology of nomogram construction.

Acid-Base Equilibrium↗

PubMed1987

Homeostatic regulation of airway smooth muscle tone by catecholamine secretion in swine.

We studied the homeostatic secretory response of catecholamine secretion elicited by progressive bronchoconstriction in 18 swine in vivo. The potential reserve of the sympathetic nervous system (SNS) was first assessed by exogenous nicotinic stimulation with 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP). A dose of 250 micrograms/kg iv DMPP caused an increase in plasma norepinephrine (NE) concentration from 207 +/- 86 (basal) to 2,625 +/- 448 pg/ml (P less than 0.02) and in plasma epinephrine (EPI) from 10 +/- 5.0 to 1,410 +/- 432 pg/ml (P less than 0.05) in four swine. In four other swine, bronchoconstriction induced by aerosolized prostaglandin F2 alpha caused approximately a fivefold increase in airway resistance without hemodynamic changes. No increase in plasma EPI was observed. However, plasma NE increased from 330 +/- 131 to 1,540 +/- 182 pg/ml (P less than 0.02). In five swine receiving aerosolized acetylcholine (ACh), similar changes in airways resistance were not associated with significant changes in catecholamine concentration when mean arterial blood pressure (MAP) was unchanged. However, inhalation of sufficient ACh to cause a greater than 10% decrease in MAP caused progressive increase in catecholamine secretion. Plasma EPI increased from 32 +/- 16 (MAP = 124 +/- 7 Torr) to 1,165 +/- 522 pg/ml (MAP = 94 +/- Torr). Hypoxemia that occurred with bronchoconstriction (greater than or equal to 50 Torr) did not cause catecholamine secretion. However, severe hypoxemia (PO2 less than 30 Torr) caused large increases in plasma EPI concentrations from 84 +/- 27 to 1,463 +/- 945 pg/ml (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗

PubMed1989

Effects of vasomotor- and mediator-induced hypotension on bronchomotor tone in swine.

We studied the sympathetic neural response on airways to hypotensive stimuli in 19 swine in vivo. The effects of pharmacologically induced hypotension with nitroprusside (NTP) and hypotension elicited by intravenous compound 48/80 (48/80), a mast cell degranulating agent, were compared after equivalent reductions in mean arterial blood pressure (MAP). Reduction of the MAP to 60% of base line with NTP in six swine caused an increase in plasma epinephrine (E) from 60 +/- 28 to 705 +/- 276 pg/ml (P = 0.032) and plasma norepinephrine (NE) from 270 +/- 46 to 796 +/- 131 pg/ml (P = 0.032). Comparable reduction in MAP elicited with 48/80 in six other swine caused a substantially greater increase in both plasma E (9,581 +/- 4,147 pg/ml; P = 0.012 vs. NTP group) and plasma NE (2,239 +/- 637 pg/ml; P = 0.041 vs. NTP group). Catecholamine secretion attenuated mediator-induced changes in lung resistance (RL). In animals receiving 48/80, RL increased from 2.97 +/- 0.31 to 7.44 +/- 0.56 cmH2O.l-1.s. In animals having ganglionic blockade with 7.5 mg/kg iv hexamethonium and beta-adrenergic blockade with propranolol (4.0 mg/kg iv followed by 40 micrograms/kg-1.min-1), comparable doses of 48/80 caused an increase in RL to 18.6 +/- 4.55 cmH2O.l-1.s (P less than 0.04 vs. swine receiving neither hexamethonium nor propranolol).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1989

Expression of airway contractile properties and acetylcholinesterase activity in swine.

We studied the effect of maturation on contractile properties of tracheal smooth muscle from seventeen 2-wk-old swine (2ws) and fifteen 10-wk-old swine (10ws) in situ and in vitro. The response to parasympathetic stimulation was studied in situ in isometrically fixed segments. Contraction was elicited at lower frequencies [half-maximal response to electrical stimulation (ES50) = 6.7 +/- 0.05 Hz] in 2ws than in 10ws (ES50 = 9.1 +/- 0.4 Hz; P less than 0.01). Despite substantial differences in morphometrically normalized cross-sectional area in 2ws (0.012 +/- 0.003 cm2) and 10ws (0.028 +/- 0.001 cm2; P less than 0.01), maximal active tension elicited by parasympathetic stimulation was similar (12.4 +/- 3.2 g/cm in 2ws vs. 13.3 +/- 2.3 g/cm in 10ws; P = NS). In separate in vitro studies in 25 tracheal smooth muscle strips from 10 swine, concentration-response curves generated with potassium-substituted Krebs solution (KCl) were similar in 2ws and 10ws. In 58 other strips (10 swine), maximal active force elicited with acetylcholine (ACh) in 2ws was significantly greater than for 10ws (P less than 0.001). Removal of the epithelium had no effect. However, cholinesterase inhibition with 10(-7) M physostigmine augmented the response to ACh in 10ws (P less than 0.02) but not 2ws. We demonstrate increased force generation and sensitivity to vagal stimulation in 2ws vs. 10ws, which corresponds to increased reactivity to ACh in vitro. The relative hyperresponsiveness in 2ws is specific for cholinergic response and is attenuated at least in part by maturation of the activity of acetylcholinesterase enzyme.

Acetylcholinesterase↗

PubMed2001

Alignment of VIM, MRC1, GAD2, and IL2RA genes on swine chromosome 10q by in situ hybridization and RH mapping.

Since the distal half of swine chromosome (SSC) 10q was shown to contain a quantitative trait locus (QTL) influencing swine growth, the precise correspondence between this chromosome region and the orthologous human chromosome region (HSA10p) was investigated using chromosomal fluorescence in situ hybridization and RH mapping of type I loci spanning the growth QTL. The goal was to align this critical region of swine with the corresponding region in human for the purpose of identifying candidate genes. The HSA10p type I loci mapped in swine were VIM, MRC1, GAD2, and IL2RA. Locus order on SSC10q was shown to be centromere-VIM-MRC1-GAD2-IL2RA, while in human the order is centromere-GAD2-MRC1-VIM--IL2RA, indicating that the chromosome segment marked by VIM, MRC1 and GAD2 has been inverted relative to the centromere and IL2RA.

Animals↗

PubMed2002

Construction of a dense comparative map between human chromosome 1p36-->p35 and swine chromosome 6 by using human sequence-tagged sites.

Construction of a comprehensive comparative map between swine and human chromosomes is a prerequisite, in order to select candidate swine genes for traits from the human genome database as well as to understand the evolutionary process of the two species. The present study attempted to use 910 sequence-tagged sites (STSs) localized in human chromosome (HSA) 1p36-->p35 (35 Mbp) for radiation hybrid (RH) mapping to swine chromosomes (SSCs). Out of the 910 STSs subjected to amplification of swine orthologues, primer pairs for 13 STSs were found to amplify the respective orthologues and the STSs were assigned to SSCs. Eleven STSs were assigned to SSC6 in the same order as that in HSA1: SSC6cen-(SHGC-150)-(A006H31)-(X82877)-(A007E03)-(IB404)-(stGDB:371372)-(stSG31658)-(A009Q18)-(stSG14201/A009C01)-(H08335)-qter. One of the remaining two STSs, WI-20819, was assigned to SSCX, and the other, R91D18R, was not linked to any first-generation markers of the IMpRH map with a lod score greater than 3.

Animals↗

PubMed2003

Elucidation of correspondence between swine chromosome 4 and human chromosome 1 by assigning 27 genes to the ImpRH map, and development of microsatellites in the proximity of 14 genes.

Loci affecting swine intramuscular fat content, backfat thickness, carcass weight, and daily weight gain were assigned to regions of swine chromosome (SSC) 4, which were shown to correspond to human chromosome (HSA) 1p22--> q25 by ZOO-FISH, bidirectional chromosome painting, as well as by the linkage map of genes. In order to select candidate genes responsible for the above traits from the human genome database, precise correspondence between SSC4 and HSA1 is a prerequisite. In the present study, 27 genes, PTGFR, GBP1, GBP2, GFI1, GCLM, ABCD3, EXTL2, KCNA3, ADORA3, KCND3, WNT2B, NRAS, SYCP1, PTGFRN, IGSF2, NOTCH2, S100A10, SHC1, SSR2, LMNA, CCT3, CD5L, PEA15, FCER1G, EAT2, DDR2, and LAMB3, located in the HSA1 region corresponding to SSC4 or possibly SSC4, were assigned to the IMpRH map. The alignment of genes from centromere to telomere in the SSC4 q arm is basically conserved in HSA1p22-->q25 with the direction from the q arm to the p arm, which is in good agreement with results from linkage mapping. In addition, the present study first demonstrated that WNT2B residing in the middle of the HSA1 region was assigned to SSC18 with a high lod score (> 5), and that at least three intrachromosomal rearrangements occurred in the region in the process of swine and human evolution. PTGFR, and LAMB3 localized at both ends of the HSA1 region were assigned to SSC6 and SSC9, respectively, which is consistent with regional correspondence reported earlier. In the course of the above analysis, microsatellite markers were developed in the proximity of eleven genes localized on SSC4, and three genes on other swine chromosomes.

Animals↗

PubMed2003

Molecular cloning and chromosomal assignment to SSC12p13-->p11 of swine chemokine receptor CCR7.

We cloned a gene encoding the swine chemokine (C-C motif) receptor 7 (CCR7) and clarified its genomic structure and chromosomal assignment. The ORF and deduced amino-acid sequence were highly conserved with human and mouse CCR7. The swine CCR7 gene was mapped to SSC12p13-->p11 by FISH analysis. Stimulation of swine peripheral blood mononuclear cells by IL-12 and IL-18, considered potent inducers of Th1 cells from analyses in humans and mice, downregulated the expression of CCR7. This is the first report of the molecular cloning, chromosomal assignment and characterization of a chemokine receptor in swine.

Amino Acid Sequence↗

PubMed1995

Assignment of the glucose transporter 1 gene (SLC2A1) to swine chromosome 6q34-->qter.

A cosmid clone containing the glucose transporter gene (SLC2A1) was selected from a cosmid library of swine genomic fragments by colony hybridization using the cDNA of swine glucose transporter as a probe. The genomic fragment thus cloned was partially sequenced, and the sequence was compared first with the sequence of the cDNA and then with the reported rat genomic sequence. The sequence homology and exon/intron structure indicated that the genomic fragment contained at least part of SLC2A1. The fragment, together with vector DNA, was then labeled by biotin and used as a probe for in situ hybridization on swine chromosome spreads. Hybridization was visualized by the FITC-labeled streptavidin/biotinylated anti-streptavidin system together with the R-band pattern of chromosomes. The fluorescence signals revealed that SLC2A1 is localized on swine chromosome 6q34-->qter.

Animals↗

PubMed1997

Swine centromeric DNA repeats revealed by primed in situ (PRINS) labeling.

In swine, distinct centromeric satellite DNA families have been described that correspond to either all the metacentric chromosomes except the Y (Mc1) or all the acrocentric chromosomes (Ac2). Using primed in situ (PRINS) labeling, we show here that primers derived from various sequences specifically label the centromeres of different subgroups of chromosomes. Among five primers derived from centromeric sequences of acrocentric chromosomes reported to be very homogeneous, four recognize all the acrocentric chromosomes, whereas one labels prominently chromosome 17. For the metacentric chromosomes, six primers have been derived from several divergent sequences. Among these primers, two recognize all the metacentric chromosomes except 5, 10, and 12. Three other primers label small subsets of metacentric chromosomes, including the X and one or two additional chromosomes. The last primer is specific to chromosome 1. These preliminary results suggest that it should be possible to define specific primers for almost every swine chromosome. Already, some of the primers reported here permit a distinction between swine chromosomes difficult to differentiate without banding, such as the X chromosome and chromosome 9. Therefore, the PRINS technique using centromeric motifs constitutes an additional tool for cytogenetic studies in swine.

Animals↗