Porcine Lung Tissue Research Articles

Addition of seaweed (Laminaria digitata) extracts containing laminarin and fucoidan to porcine diets: Influence on the quality and shelf-life of fresh pork

A seaweed extract containing laminarin (L) and fucoidan (F) (L/F) was manufactured from brown seaweed (Laminaria digitata) in spray-dried (L/F-SD) and wet (L/F-WS) forms. The effect of supplementation of pig diets with L/F-SD and L/F-WS (L, 500mg/kg feed; F, 420mg/kg feed) for 21days pre-slaughter, on quality indices of fresh M. longissimus dorsi (LD) steaks was examined. Susceptibility of porcine liver, heart, kidney and lung tissue homogenates to iron-induced (1mM FeSO4) lipid oxidation was also investigated. Dietary supplementation with L/F did not increase plasma total antioxidant status (TAS). In LD steaks stored in modified atmosphere packs (80% O2:20% CO2) (MAP) for up to 15days at 4°C, muscle pH, surface colour (CIE ‘L*’ lightness, ‘a*’ redness and ‘b*’ yellowness values) and microbiology (psychrotrophic and mesophilic counts, logCFU/g pork) were unaffected by dietary L/F. In general, levels of lipid oxidation (TBARS, mg MDA (malondialdehyde)/kg pork) followed the order: C>LF-SD>L/F-WS. A statistically significant reduction in lipid oxidation (P<0.05) was observed in LD steaks from 75% of pigs (n=6) fed with L/F-WS compared to controls. Iron-induced lipid oxidation increased in liver, heart, kidney and lung tissue homogenates over the 24h storage period and dietary L/F-WS reduced lipid oxidation to the greatest extent in liver tissue homogenates. Results demonstrate potential for the incorporation of marine-derived bioactive antioxidant components into muscle foods via the animal's diet.

Pharmacokinetics of tildipirosin in porcine plasma, lung tissue, and bronchial fluid and effects of test conditions on in vitro activity against reference strains and field isolates of Actinobacillus pleuropneumoniae

The pharmacokinetics of tildipirosin (Zuprevo(®) 40 mg/mL solution for injection for pigs), a novel 16-membered-ring macrolide for the treatment for swine respiratory disease (SRD), was investigated in studies collecting blood plasma and postmortem samples of lung tissue and bronchial fluid (BF) from swine. In view of factors influencing the in vitro activity of macrolides, and for the interpretation of tildipirosin pharmacokinetics in relation to minimum inhibitory concentrations (MIC), additional experiments were conducted to study the effects of pH, carbon dioxide-enriched atmosphere, buffers, and serum on tildipirosin MICs for various reference strains and Actinobacillus (A.) pleuropneumoniae field isolates. After single intramuscular (i.m.) injection at 4 mg/kg body weight, maximum plasma concentration (Cmax) was 0.9 μg/mL observed within 23 min (Tmax ). Mean residence time from the time of dosing to the time of last measurable concentration (MRTlast) and terminal half-life (T1/2) both were about 4 days. A dose-response relationship with no significant sex effect is observed for area under the plasma concentration-time curve from time 0 to the last sampling time with a quantifiable drug concentration (AUClast) over the range of doses up to 6 mg/kg. However, linear dose proportionality could not be proven with statistical methods. The time-concentration profile of tildipirosin in BF and lung far exceeded that in blood plasma. In lung, tildipirosin concentrations reached 3.1 μg/g at 2 h, peaked at 4.3 μg/g at day 1, and slowly declined to 0.8 μg/g at day 17. In BF, tildipirosin levels were 14.3, 7.0, and 6.5 μg/g at days 5, 10, and 14. T1/2 in lung was ∼7 days. Tildipirosin is rapidly and extensively distributed to the respiratory tract followed by slow elimination. Culture media pH and carbon dioxide-enriched atmosphere (CO2 -EA) had a marked impact on in vitro activity of tildipirosin in reference strains of various rapidly growing aerobic and fastidious bacteria including Histophilus (H.) somni ATCC 700025 and A. pleuropneumoniae ATCC 27090. For A. pleuropneumoniae ATCC 27090 testing conditions without CO2 -EA resulted in reduced acidification of culture media pH and a reduction in the minimum inhibitory concentrations compared to standard in vitro test conditions by 2 log2 dilution steps (4-fold) from 8 to 2 μg/mL. Supplementary buffering of standard culture media resulted in a reduction in the A. pleuropneumoniae (n = 8) MIC range by 4 log2 dilution steps (16-fold) from 8-16 to 0.5-1 μg/mL. Incremental supplementation of culture media with 50% serum resulted in noticeable shifts to lower minimum or maximum MICs by at least 2 log2 dilution steps (≥4-fold) in all aerobic and fastidious reference strains tested except for Pasteurella (P.) multocida. The MIC of A. pleuropneumoniae ATCC 27090 decreased by 2-4 log2 dilution steps (4 to 16-fold) from 8 to 0.5-2 μg/mL when 50% serum was added to the standard assay. Considering a higher presence of serum and the rather neutral pH conditions maintained in vivo, it is suggested to take the influence of these factors on in vitro activity into account when interpreting tildipirosin MICs for A. pleuropneumoniae in relation to pharmacokinetics.

Different expression patterns of PRRSV mediator genes in the lung tissues of PRRSV resistant and susceptible pigs

Porcine reproductive and respiratory syndrome (PRRS) has caused severe economic loss in most swine-producing countries. The resistance to PRRS virus (PRRSV) infection varies among pig breeds and lines. In this study, we found that the Chinese Dapulian pigs (DPL) were more resistant to PRRSV than commercial Duroc×Landrace×Yorkshire (DLY) crossbred pigs in that lower rectal temperature and lower PRRSV copy number in the serum were detected in the former. Analysis of the mRNA expression of five PRRSV mediator genes (SIGLEC1, NMMHC-IIA, CD163, VIM and HSPG2) in the lung tissues indicated differences in expression between DLY and DPL pigs. In uninfected porcine lung tissues, the levels of SIGLEC1, NMMHC-IIA, CD163 and VIM genes were significantly higher in DLY than in DPL pigs (P<0.05); in PRRSV-infected pigs, the expression levels of NMMHC-IIA and CD163 mRNA were significantly higher in DPL pigs compared to uninfected ones (P<0.05), whereas these levels were not different in DLY pigs or between infected DPL and DLY pigs. Thus, the different expression of PRRSV mediator genes is likely related to pig resistance to PRRSV.

Oligopeptide transport and regulation of extracellular proteolysis are required for growth of Aspergillus fumigatus on complex substrates but not for virulence

Moulds are characterized by their saprophytic lifestyle that is based on osmotrophy. Among them, Aspergillus fumigatus has emerged as the leading cause of fungal infections in the presence of an underlying immunodeficiency. To assess the role of its nutritional versatility for virulence, transcriptional profiling studies in the presence of varying sources of nitrogen were carried out and revealed an extensive reprogramming of the fungal transcriptome when shifting to a proteinaceous growth substrate. Transcripts encoding metabolic activities were predominantly upregulated, as were proteinases and transport activities. To probe whether fundamental aspects of its osmotrophic lifestyle, that is, extracellular proteolysis and uptake of oligopeptides, are required for A. fumigatus pathogenicity, serial gene replacements were carried out, which eventually yielded an octuple deletion mutant ablated for the opt gene family. This strain displayed no growth defect on various substrates, but supplementary reduction of extracellular proteolytic activity by additional deletion of the prtT gene revealed a synthetic phenotype on porcine lung tissue agar. Virulence studies in a murine model of pulmonary aspergillosis did not disclose any attenuation in virulence of these deletants. Our data emphasize a high degree of redundancy encoded by the A. fumigatus genome that secures nutrient supply for growth and, therefore, virulence.

Anti-inflammatory, antinociceptive and anti-angiogenic activities of a phospholipid mixture purified from porcine lung tissues

This work aimed to assess anti-inflammatory and related properties of a phospholipid mixture purified from porcine lung tissues, named KT&G101, which is being developed as a novel topical remedy for atopic dermatitis. KT&G101 consists of pure phospholipids, mainly phosphatidylcholine (PC) and other phospholipids such as phosphatidylinositol (PI) and phosphatidylserine (PS). Its predominant PC species is 1,2-dipalmitoylphosphatidylcholine (DPPC). KT&G101 exhibited an anti-angiogenic activity in the chick chorioallantoic membrane (CAM) assay. Oral administration of KT&G101 at the dosages of 100, 200 and 400 mg/kg body weight gave rise to an inhibition of 15.4%, 25.3% and 30.1% in the vascular permeability assay, respectively. In the carrageenan-induced inflammation in the air pouches, KT&G101 significantly diminished the volume of exudates in the pouches, the number of polymorphonuclear leukocytes and nitrite content in exudates. In the acetic acid-induced writhing response, oral administration of KT&G101 at the dosages of 50, 100 and 200 mg/kg body weight showed the reduction of 21.6%, 51.6% and 60.8% in the pain response of mice, respectively. It was also able to diminish the nitric oxide (NO) and reactive oxygen species (ROS) levels in the lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. KT&G101 displayed a significant suppression on the induction of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in the stimulated RAW264.7 cells. However, the free radical scavenging activity of KT&G101 was detected to be very weak in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. Taken together, KT&G101 possesses anti-inflammatory and related antinociceptive and anti-angiogenic activities, which indirectly supports its use as an anti-atopic therapy.

The role of vascular adhesion molecules PECAM-1 (CD 31), VCAM-1 (CD 106), E-selectin (CD62E) and P-selectin (CD62P) in severe porcine pancreatitis.

Inflammatory cytokines have been shown to mediate organ damage by their action on vascular endothelia and leukocytes, in part by upregulating the expression of adhesion molecules, which in turn convey transmigration of leukocytes into tissue. The upregulation and activation of vascular cell adhesion molecules on the endothelial cells avail firm leukocyte adhesion to the vascular endothelium and enhance their transmigration and consecutive tissue injury. The aim of this study was to evaluate the expression of vascular adhesion molecules CD 31 (PECAM-1), CD 106 (VCAM-1), CD 62E (E-Selectin) and CD 62P (P-Selectin) in the pancreas and distant organs of pigs suffering from acute necrotizing pancreatitis (AP). AP was induced in 13 pigs by a combination of intravenous cerulein and intraductal glycodeoxycholic acid. For immunostaining of vascular adhesion molecules slides of porcine pancreas, lung, kidney and liver tissue were stained with monoclonal antibodies (Ab) against PECAM-1-1, VCAM-1 E- and P- SELECTIN. The endothelial cell expression of CD 31 (PECAM-1), CD 106 (VCAM), CD 62E (E-Selectin) and CD 62P (P-SELECTIN) in severe porcine pancreatitis is detectable and upregulation is partly significantly.

An isogenic Actinobacillus pleuropneumoniae AasP mutant exhibits altered biofilm formation but retains virulence

AasP, an autotransporter serine protease of Actinobacillus pleuropneumoniae, has been shown to be expressed in necrotic porcine lung tissue. Based on the hypothesis that AasP might play an important role in A. pleuropneumoniae adhesion and virulence by processing other surface-associated proteins, the predicted catalytic site of AasP was deleted and the isogenic mutant, AP76Δ aasP, was compared to the wild-type strain in a biofilm assay as well as an aerosol infection model. AP76Δ aasP showed increased adherence compared to the wild-type strain under standard culturing conditions as well as under NAD restriction. No significant differences between AP76 wild-type and AP76Δ aasP were observed upon experimental infection of pigs, indicating that AasP does not play a crucial role in A. pleuropneumoniae virulence.

Identification of genes transcribed by Haemophilus parasuis in necrotic porcine lung through the selective capture of transcribed sequences (SCOTS)

Haemophilus parasuis is the aetiological agent of Glässer's disease, which has received more attention in the past decade due to the increasing economic losses in the pig industry worldwide. Little is known about the mechanisms by which H. parasuis survives in the host. In this study, selective capture of transcribed sequences (SCOTS) was used to identify H. parasuis genes upregulated in necrotic porcine lung 7 days post infection. Thirty-eight genes were identified that were upregulated during infection of the lung tissue of pigs, compared with growth in culture medium. In two examples chosen gene expression was not confined to the lungs, there being variation between tissues. The data support biofilm formation being an important mode of growth for colonization and/or persistence. Results from the in vitro studies suggest that, as for other pathogens, iron and oxygen restriction and heat stress are important environmental signals to regulate gene expression. This study has identified genes of H. parasuis that are upregulated during infection of porcine lung tissue as compared with in vitro growth conditions.

Selective capture of transcribed sequences (SCOTS) of Actinobacillus pleuropneumoniae in the chronic stage of disease reveals an HlyX-regulated autotransporter protein

Actinobacillus pleuropneumoniae, an important respiratory pathogen in swine, is able to persist in host tissues for extended periods of time. In the study presented here, selective capture of transcribed sequences (SCOTS) analysis was used to identify genes expressed by A. pleuropneumoniae in the chronic stage of the disease (21 days post infection). After isolation and reverse transcription of RNA from infected lungs as well as from culture-grown A. pleuropneumoniae, transcribed A. pleuropneumoniae sequences were captured from infected lung tissue and subjected to a subtractive hybridization procedure of lung-derived against culture-derived A. pleuropneumoniae cDNA. Twenty-nine of the thirty-six genes that were identified as in vivo-expressed are involved in transport or metabolic processes. We identified a surface-associated putative 104 kDa subtilisin-like autotransporter serine protease, designated AasP, which has not been described in A. pleuropneumoniae to date. The gene was shown to be present in all 15 A. pleuropneumoniae serotypes. It is transcribed in porcine lung tissue on days 7 and 21 post infection. Under anaerobic conditions in vitro, its expression depends on the global anaerobic regulator HlyX. To our knowledge, this is the first report of an autotransporter protein being regulated by a global anaerobic regulator.

Detection of respiratory pathogens in porcine lung tissue and lavage fluid

The objective of this study was to compare the detection rate of bacterial agents in bronchoalveolar lavage fluid (BALF), taken without visual control, to that in affected lung tissue obtained from the same pig at necropsy. BALF and affected lung tissue were examined for Mycoplasma hyopneumoniae using PCR, and standard cultural methods were used for Actinobacillus pleuropneumoniae, Bordetella bronchiseptica, Haemophilus parasuis, Pasteurella multocida and Streptococcus suis. All pigs with a history of respiratory symptoms were submitted as live animals for routine diagnostic examination. In each animal the site of lavage, marked by injecting methylene blue, differed from the site of pneumonic lesions. M. hyopneumoniae was detected more frequently in lung tissue than in BALF in cases with moderate or severe lung lesions. The detection rates of M. hyopneumoniae were higher in the BALF of pigs with mild lesions. Cultural examination of BALF was at least as satisfactory as affected lung tissue for detecting B. bronchiseptica, H. parasuis and P. multocida.

Application and Field Validation of a PCR Assay for the Detection of Mycoplasma Hyopneumoniae from Swine Lung Tissue Samples

A PCR assay was validated for the detection of Mycoplasma hyopneumoniae in porcine lung tissue. The detection limit of the assay was 0.18 colony-forming units/g of lung sample spiked with M. hyopneumoniae. In field validation, 426 pigs from 220 cases were examined for M. hyopneumoniae infection by M. hyopneumoniae PCR and a fluorescent antibody (FA) test. In total, 103 pig lungs (24.2%) were positive in the PCR test, and 69 pig lungs (16.2%) were positive in the FA test, among which, 62 pigs were positive for both PCR and FA test. Most of the PCR-positive but FA test-negative cases had lesions compatible with M. hyopneumoniae infection. With Bayesian modeling, the diagnostic sensitivity and specificity of the PCR were determined to be 97.3% and 93.0%, respectively.

325: Histopathologic Effect of Endotracheal Drug Administration on Porcine Lung Tissue

325: Histopathologic Effect of Endotracheal Drug Administration on Porcine Lung Tissue

Gene expression profiling in Salmonella Choleraesuis-infected porcine lung using a long oligonucleotide microarray

Understanding the transcriptional response to pathogenic bacterial infection within food animals is of fundamental and applied interest. To determine the transcriptional response to Salmonella enterica serovar Choleraesuis (SC) infection, a 13,297-oligonucleotide swine array was used to analyze RNA from control, 24-h postinoculation (hpi), and 48-hpi porcine lung tissue from pigs infected with SC. In total, 57 genes showed differential expression (p < 0.001; false discovery rate = 12%). Quantitative real-time PCR (qRT-PCR) of 61 genes was used to confirm the microarray results and to identify pathways responding to infection. Of the 33 genes identified by microarray analysis as differentially expressed, 23 were confirmed by qRT-PCR results. A novel finding was that two transglutaminase family genes (TGM1 and TGM3) showed dramatic increases in expression postinoculation; combined with several other apoptotic genes, they indicated the induction of apoptotic pathways during SC infection. A predominant T helper 1-type immune response occurred during infection, with interferon gamma (IFNG) significantly increased at 48 hpi. Genes induced by IFNs (GBP1, GBP2, C1S, C1R, MHC2TA, PSMB8, TAP1, TAP2) showed increased expression during porcine lung infection. These data represent the first thorough investigation of gene regulation pathways that control an important porcine respiratory and foodborne bacterial infection.

Gene expression of pulmonary cytokines after sevoflurane or thiopentone anaesthesia in pigs

Volatile anaesthetics have diverse inflammatory effects on the lungs. They increase gene expression of some pro-inflammatory cytokines in alveolar macrophages whereas in alveolar type II cells they seem to decrease secretion and gene expression of pro-inflammatory cytokines. We have previously detected increased leukotriene C4, nitrate and nitrite concentrations in bronchoalveolar lavage fluid after sevoflurane anaesthesia. In the current study, we measured gene expression of inflammatory cytokines in the lung tissue and plasma concentrations of cytokines in pigs after thiopentone or sevoflurane anaesthesia. Sixteen pigs were randomly selected to receive either a continuous thiopentone infusion (control group, n = 8) or sevoflurane (n = 8) at 4.0% inspiratory concentration (1.5 MAC) in air for 6 h. Tissue samples were collected at the end of the study for measurement of gene expression of inflammatory cytokines. Blood samples were collected during anaesthesia for measurement of plasma cytokine concentrations. Compared with thiopentone anaesthesia, lower gene expression of tumour necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) in lung tissue was observed after sevoflurane anaesthesia. Of measured cytokines IL-1beta, TNF-alpha, IL-6, IL-8 and IL-10 only plasma concentrations of IL-6 could be measured during the study without a difference between the groups. Lower gene expression of TNF-alpha and IL-1beta was found in the intact porcine lung tissue after sevoflurane anaesthesia compared with thiopentone anaesthesia. Clinical significance of this finding is unknown.

Identification of Genes Transcribed byActinobacillus pleuropneumoniaein Necrotic Porcine Lung Tissue by Using Selective Capture of Transcribed Sequences

Genes expressed by Actinobacillus pleuropneumoniae in necrotic porcine lung tissue were identified by selective capture of transcribed sequences analysis. In total, 46 genes were identified, 20 of which have been previously reported to be associated with in vivo expression or virulence in A. pleuropneumoniae or in other organisms.

Reactivity of Anti-Nipah Virus Monoclonal Antibodies to Formalin-fixed, Paraffin-embedded Lung Tissues from Experimental Nipah and Hendra Virus Infections

The immunohistochemical reactivity of seven clones of mouse monoclonal antibodies raised to Nipah virus antigens were investigated using formalin-fixed, paraffin embedded porcine and equine lung tissues from experimental Nipah and Hendra virus infection, respectively. Either microwave irradiation or enzymatic digestion effectively unmasked the viral antigens in formalin-fixed, paraffin-embedded tissue sections. Four clones showed positive reaction to both Nipah virus-infected porcine lung tissue and Hendra virus-infected equine lung tissue. Two clones (11F6 and 13A5) reacted with Nipah virus-infected porcine lung tissue, but not with Hendra virus-infected equine lung tissue. These Nipah virus-specific monoclonal antibodies may therefore be useful for immunohistological diagnosis of Nipah virus infection and for further research on Nipah virus pathogenesis.

Characterization of the porcine melanocortin 2 receptor gene (MC2R ).

A porcine bacterial artificial chromosome (BAC) clone, containing the melanocortin 2 receptor gene (MC2R) was isolated. The complete coding sequence of the MC2R gene, contained in 1 exon, was determined. Polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) was performed on a 241-bp coding fragment. An AluI polymorphism, detecting a silent mutation, was found and typed on unrelated animals of five different pig breeds. The Meishan, Piétrain and Large White breeds differ significantly in allele frequencies from the Landrace and Czech Meat Pig breeds. The melanocortin 5 receptor gene (MC5R) was detected by PCR in the same BAC clone, as could be expected from the human and porcine mapping data. PCR-SSCP was performed on a 200-bp coding of MC5R, but no polymorphisms were detected. The BAC clone was mapped to Sscr6q27 by fluorescent in situ hybridization (FISH). A (CA)n microsatellite (SGU0002), isolated from the BAC, was localized on chromosome 6 by RH mapping near marker SW1473 and by linkage mapping on the MARC reference family at the same position as the marker SW2173 (97 cM). Allele frequencies, heterozygosity and polymorphism information contents (PIC) values were calculated for the five different pig breeds examined. The transcription of both genes in porcine liver, heart, kidney, fat, brain, pancreas, stomach, bladder, ovaries, lung, spleen, skin, adrenal gland and muscle tissues was examined by reverse transcriptase-polymerase chain reaction. Transcription was detected in skin and adrenal gland tissues for MC2R, while a positive signal was detected for MC5R in kidney, fat, pancreas, skin, adrenal gland and spleen tissues.

Aerosolized perfluorocarbon suppresses early pulmonary inflammatory response in a surfactant-depleted piglet model.

The effect of new ventilation strategies on initial pulmonary inflammatory reaction was studied in a surfactant-depleted piglet model. Sixty minutes after induction of lung injury by bronchoalveolar lavage, piglets received either aerosolized FC77 (aerosol-PFC, 10 mL/kg/h, n = 5) or partial liquid ventilation (PLV) with FC77 at functional residual capacity volume (FRC-PLV, 30 mL/kg, n = 5), or at low volume (LV-PLV, 10 mL/kg per hour, n = 5), or intermittent mandatory ventilation (control, n = 5). After 2 h, perfluorocarbon application was stopped and intermittent mandatory ventilation continued for 6 h. After a total experimental period of 8 h, animals were killed and lung tissue obtained. mRNA expression of IL-1beta, IL-6, IL-8, and TGF-beta in porcine lung tissue was quantified using TaqMan real-time PCR and normalized to beta-actin (A) and hypoxanthine-guanine-phosphoribosyl-transferase (H). In the aerosol-PFC group, IL-1beta, IL-6, IL-8, and transforming growth factor (TGF)-beta mRNA expression in lung tissue was significantly lower than in the control group. Reduction was 95% for IL-1beta/H (p < 0.001), 73% for IL-6/H (p < 0.05), 87% for IL-8/H (p < 0.001), and 38% for TGF-beta/H (p < 0.01). A lower mRNA gene expression was also determined for IL-1beta and IL-8 when the aerosol-PFC group was compared with the LV-PLV group [91% for IL-1beta/H (p < 0.001), 75% for IL-8/H (p < 0.001)]. In the FRC-PLV group, mRNA expression of IL-1beta was significantly lower than in the control (p < 0.05) and LV-PLV (p < 0.01) group. In a surfactant-depleted piglet model, aerosol therapy with perfluorocarbon but not LV-PLV reduces the initial pulmonary inflammatory reaction at least as potently as PLV at FRC volume.

Distribution and quantification of beta1-, beta2-, and beta3-adrenergic receptor subtype transcripts in porcine tissues.

Distribution and concentration of beta-adrenergic receptor (betaAR) subtype transcripts (beta1AR, beta2AR, and beta3AR) were investigated in porcine and rat tissues. Reverse transcription (RT) coupled with PCR indicated the presence of beta1AR and beta2AR transcripts in porcine left ventricle, lung, longissimus muscle, and subcutaneous adipose tissue. The RT-PCR indicated that beta3AR transcripts were present in porcine subcutaneous adipose tissue, and transcripts were suggested in left ventricle and lung, but they were not detected in longissimus muscle. Quantitative ribonuclease protection assays indicated that the porcine beta1AR, beta2AR, and beta3AR transcript concentrations (amol/microg of total RNA) were, respectively as follows: heart left ventricle = 6.1, 2.4, and .02; lung = 3.8, 1.9, and .01; liver = 1.7, 2.1, and undetectable; skeletal muscle = 1.7, 1.1, .02; subcutaneous adipose tissue = 1.3, .4, .14. The proportions of porcine betaAR subtypes (beta1AR:beta2AR:beta3AR) were as follows: heart left ventricle = 72:28:.25; lung = 67:33:.2; liver = 45:55:0; skeletal muscle = 60:39:.7; and subcutaneous adipose = 73:20:7. Normalization of data to beta-actin transcript concentration did not change these relationships. Rat perigonadal adipose tissue beta1AR and beta3AR transcript concentrations were, respectively, .59 and 1.84 amol/microg of total RNA. The rat perigonadal adipose tissue concentration of beta3AR transcript was tenfold that of porcine subcutaneous adipose tissue. The predominant betaAR transcript in rat adipose tissue was beta3AR, and the predominant betaAR transcript in pig adipose tissue was beta1AR.

Species Difference in the Binding Characteristics of Res-701-1: Potent Endothelin ETB Receptor-Selective Antagonist

Species-dependency of RES-701-1 (cyclic (Gly1-Asp9) (Gly-Asn-Trp-His-Gly-Thr-Ala-Pro-Asp-Trp-Phe-Phe-Asn-Tyr-Tyr-Trp)) has been shown using 125I-ET-1 binding to membranes from various animal tissues. RES-701-1 selectively inhibited the 125I-ET-1 binding to ETB receptor from canine, rabbit, porcine, and guinea pig lung tissues in a dose-dependent manner with IC50 values of 60 nM, 20, 4 nM and 13 nM, respectively. Though RES-701-1 retained selectivity for ETB receptor even at higher concentrations, RES-701-1 inhibited the 125I-ET-1 binding to ETB receptor from rat lung, heart, and kidney only weakly; IC50 values were calculated to be 1.2 μM, 0.9 μM and 0.6 μM, respectively. These results suggest that RES-701-1 is a potent and specific antagonist for ETB receptor in various species of animal, and would be a powerful tool for understanding the physiological roles of ETB receptor, but care must be taken in use of RES-701-1 in experiments with rat.