Immunohistochemical localization of S-100 protein in the pig pituitary gland

Immunohistochemical localization of S-100 protein was studied in anterior, intermediate and posterior lobe of the pig pituitary gland. Two immunopositive cells for S-100 protein were identified: the folliculo-stellate cells (FSc) in the glandular lobes and the pituicytes in the neural lobe. In the anterior lobe, immunoreactive folliculo-stellate cells were scattered among secretory cells. In the area where the secretory cells form strands and follicle-like groups the positive cells were concentrated in groups. In the intermediate lobe, S-100 protein-positive cells were located sparsely among secretory cells and next to secretory follicles and the pituitary cleft. These FSc were more voluminous and displayed fewer cytoplasmic processes. In the neurohypophysis, positive reaction for S-100 protein was seen in the pituicytes. These cells were distributed singly or concentrated in groups. The distribution, and morphologic characteristics of the FSc in the glandular lobes and the pituicytes in the neural lobe in the pig indicate different origin of both S-100 protein-positive cells.     

Characterisation of sugar residues in glycoconjugates of pig mandibular gland by traditional and lectin histochemistry

Sugar residues are important components of salivary gland secretion. Traditional histochemical methods and lectin histochemistry were used to characterise glycoconjugates present in the mandibular gland of normal adult pigs. Acinar cells contained abundant quantities of glycoconjugates with the terminal trisaccharide sialic acid - (alpha 2-->3, 6) galactosyl (beta 1-->3) N -acetylgalactosamine. Mandibular acinar cells also contained alpha and beta N -acetylgalactosamine and N -acetylglucosamine residues, whereas the demilunar cells contained glycoconjugates with fucose, mannose and N -acetylglucosamine residues. In the duct system a range of sugar residues were localised throughout the cell cytoplasm or limited to the apical surface. These results provide new knowledge concerning the structure of salivary glycoconjugates in normal adult pig and a basis for future pathological studies.     

Immunophenotypic characterization of peripheral blast cells in a leukemic miniature pig

The health status of a 4-year-old female, dd-haplotype miniature pig deteriorated rapidly, so the animal finally had to be euthanized because of poor clinical condition. Necropsy revealed a massive leukocytic infiltration in the parenchymatous organs of the abdominal cavity. On hematologic cell counting, severe leukocytosis (69.3 x 10(9) cells/liter) and high-grade basophilia (6.9 x 10(9) cells/liter) were evident. Cytologic examination, as well as analysis of expression of leukocyte differentiation antigens by means of flow cytometry, classified blasts, which accounted for about 22% of leukocytes, as biphenotypic cells co-expressing the myeloid marker SWC3 (CD172a) and the lymphoid markers CD5 and CD25. Hematologic features resembled those seen in humans with chronic myeloid leukemia at blast phase.     

Hematologic values of the yucatan miniature pig

Mean values and ranges for 16 hematological parameters for healthy, young, sexually-mature Yucatan miniature swine are presented. No significant differences were observed between sexes with regard to hemograms. Comparison of observed values with those published in the literature for standard domestic and other breeds of miniature swine reveals no major differences.     

Improved isolation and culture of embryonic stem cells from Chinese miniature pig

Pigs serve as a better research model for human beings than other species. The Chinese laboratory miniature pig is a new laboratory animal and is expected to be applicable in many medical research fields. This study was to establish effective technologies to isolate and culture ES cells in Chinese miniature pigs. For isolation of the inner cell mass from blastocysts, an enzyme-digestive method was compared with the traditional immunosurgery. Isolated ICM were cultured in three feeder cell layers: mouse embryonic fibroblasts (MEF), porcine embryonic fibroblasts (PEF) and a continuous cell line of mouse embryonic fibroblasts (STO). Microtubule activity of the three feeder cells was further examined by immunofluorescence. ICM were successfully isolated from 85% of blastocysts by the enzyme-digestive method, compared to only 40% by immunosurgery. When ICM were cultured in three feeder layers for two to three days, 75%, 65% and 20% of ICMs formed primary cell colonies in MEF, PEF and STO, respectively. Colonies were also formed during subcultures after 9, 5 and 1 passage in MEF, PEF and STO, respectively. Microtubules in STO cells were significantly fewer than those in MEF and PEF. When the ES-like cells were cultured in a differentiation medium, they differentiated to neuron-like cells and other types of cells. These results indicate that healthier ICM can be obtained with the enzyme-digestive method. Successful culture of ICM to ES-like cells has been achieved not only in MEF, but also in homologous (pig) feeder layer. The ES cells obtained in the present study were pluripotent.     

Cellular localization of IL-18 and IL-18 receptor in pig anterior pituitary gland

Pro-inflammatory cytokine interleukin 18 (IL-18) has been proposed to have a role in modulating immuno-endocrine functions. Our previous study showed that IL-18 and IL-18 receptor (IL-18R) colocalized in somatotrophs of the bovine anterior pituitary gland, and the possibility that IL-18 acts on somatotrophs as an autocrine factor. In the present study, we investigated the localization of IL-18 and IL-18R in the pig anterior pituitary gland. RT-PCR analysis showed the expression of IL-18 and IL-18R mRNAin the pig anterior pituitary gland. Immunohistochemistry of IL-18 and specific hormones revealed the presence of IL-18 in somatotrophs, mammotrophs, thyrotrophs and gonadotrophs. IL-18R was localized in somatotrophs and thyrotrophs. Furthermore, the somatotrophs immunoreactive for IL-18 did not contain IL-18R. Thus, IL-18R and IL-18 were not colocalized in an identical somatotroph. These findings suggest that the localization of IL-18 in pig somatotrophs is different from that in bovine somatotrophs, although IL-18 closely associates with somatotrophs in the anterior pituitary glands in both species.     

Localization of glycoconjugates in dog parotid gland by lectin histochemistry

Parotid glands from adult dogs were stained with a battery of seven horseradish peroxidase-conjugated lectins (PNA, UEA, LTA, DBA, SBA, WGA and ConA). In some cases (PNA and DBA) neuraminidase digestion was followed by lectin staining. Acinar cells contained conspicuous quantities of oligosaccharides with terminal sialic acid radicals. Galactosil-(13)N-acetylgalactosamine was the most abundant penultimate sugar linked toN-acetylneuraminic acid. Sialylated components having the terminal dimer sialic acid-N-acetylgalactosamine were found in the acinar cells. Secretory cells presented a heterogeneous distribution of glycoconjugates with terminal fucose and -N-acetylgalactosamine. Fucose,N-acetylglucosamine and -N-acetylgalactosamine were present on the apical cytoplasm and surface of the striated and interlobular duct cells. This glycosidic composition was unaffected by extensive selective breeding. The role of abundant amounts of sialic acid radicals in the oral mucosa was considered.Abbreviations HRP horseradish peroxidase - PBS phosphate-buffered saline see also Tables I and III     

Role of intestinal epithelial cells in the innate immune defence of the pig intestine

The intestinal epithelium serves as a dynamic barrier, which in the course of its normal function, maintains regulated uptake of nutrients and water while excluding potential pathogens. Over the past decade many studies have also revealed the immunological importance of intestinal epithelial cells (IEC). IEC have developed a variety of mechanisms to reduce the risk of infection by invasive pathogens or damage by toxic compounds. The effective maintenance of a physical barrier function is dependent on the establishment of well-organised intercellular junctions and a constant state of regeneration/renewal of the epithelium. IEC also participate in the innate immune responsiveness of the intestine by their ability to secrete mucus and antimicrobial peptides. IEC are also able to secrete cytokines and to respond to exogenous chemokines. This review summarises the current knowledge of the innate immune mechanisms developed by porcine IEC.     

Localization of hyaluronic acid in the seminal vesicles of the miniature pig

We studied the detailed localization of hyaluronic acid in the seminal vesicles of the miniature pig, using hyaluronic acid-binding protein as a specific histochemical probe at the ultrastructural level. According to the results, the basolateral surface of the plasma membrane of the glandular epithelial cells, was found to contain hyaluronan. However, abundantly present was hyaluronan in the subepithelial connective tissue, in particular, in the extracellular matrix surrounding the fibroblasts, smooth muscle cells, small blood vessels and capillaries. The substance was also observed in the surface coat of the plasma membrane of the fibroblasts, but not in that of the smooth muscle cells. The findings suggest that hyaluronan in the seminal vesicles of the miniature pig is synthesized onto the surface coat of the plasma membrane of the fibroblasts, is contributed to the extracellular matrix, and consequently concentrates in the subepithelial connective tissue. The substance may particularly be involved in a variety of cellular functions to maintain morphological organization as well as to regulate physiological homeostasis in the reproductive organ of this species, rather than participate in sperm functions.     

Ghrelin对猪小肠上皮细胞增殖的影响

以体外分离培养的猪小肠上皮细胞为试验材料,通过分别添加1×10-5、1×10-6、1×10-7、1×10-8、1×10-9和1×10-10mol/L的Ghrelin液,来测定Ghrelin对猪小肠上皮细胞增殖的影响。试验结果表明,Ghrelin的浓度在1×10-7-1×10-6mol/L时能够显著促进体外培养的猪小肠上皮细胞的增殖(P<0.05),且在刺激后第5天达到增殖高峰。     

Calcium mediation of the pig jejunal secretory response.

The involvement of Ca++ ions as secretory mediators in pig jejunal epithelia has been investigated with an in vitro system. Omission of Ca++ from the Ringer-HCO3 bathing media on both sides of the tissue had minor effects on the basal electrical activity of pig jejunal mucosa. There were only slight decreases in transepithelial potential difference and increases in conductance with Ca++ free media. Low EGTA concentrations which reversibly blocked potential difference responses to secretory agents also had minimal effects on basal electrical activity. The in vitro secretory responses to A23187, to theophylline, and to Escherichia coli heat-stable enterotoxin were all eliminated by Ca++ depletion and restored by replacing normal Ca++ concentrations in the bathing media. Dantrolene prevented the secretory response but not the potential difference increases caused by heat-stable enterotoxin and A23187, suggesting that intracellular Ca++ stores may be reservoirs of secretory signal agent. Verapamil only blocked the secretory response to heat-stable enterotoxin. Chlorpromazine had negligible effects on basal conditions, but totally blocked both the secretory response and the Ca++-dependent effects of A23187 and heat-stable enterotoxin on potential difference. The response to theophylline was only partially inhibited by chlorpromazine, implying some involvement of both cAMP and Ca++ as secretory signals for theophylline. Cytoplasmic Ca++ concentrations appear to be at least as important as cyclic nucleotides in regulating the secretory effects of pig jejunum.     

贵州小型猪精子显微结构研究

应用光学显微镜、透射电镜和扫描电镜对贵州小型猪精子的形态和结构进行了研究.结果表明,贵州小型猪精子由头、尾二部分组成.精子全长57.00±1.69 μm;头部呈椭圆形体,长为9.49±0.40 μm;尾部由颈、中、主和末段组成;尾部中央为9×2 2排列模式的轴丝,轴丝外围依次是外周致密纤维、线粒体螺旋鞘、纤维鞘、细胞膜;线粒体螺旋鞘仅存在于中段,外周致密纤维从主段到末段逐渐变细,并在主段近末端处分批终止,末段外围仅保留纤维鞘、细胞膜.贵州小型猪精子组织结构与人类接近,是研究人类男性生殖系统疾病较理想的动物模型.     

Factors influencing the degree of in vitro bacterial adhesion to ovine mammary gland epithelial cells

Bacterial adhesion to mammary gland epithelial cells (EC) may play a role in the pathogenesis of mastitis. In vitro adherence systems have been developed to study mastitis in cattle but little has been done in sheep. In this work, a method is described for obtaining mammary gland cell preparations containing greater than or equal to 65% EC from live or dead ewes, using a Ficoll-Hypaque flotation method (cell viability = 70-90%). An in vitro adhesion assay procedure was also developed to study the interaction between EC and ovine mastitis bacterial strains. It was observed that, under the test conditions, adherence increased as the incubation time was prolonged from 30 to 120 min (P less than 0.05). Adhesion was greater at incubation temperature of 37 degrees C than at 22 degrees C (P less than 0.001). An acidic pH (5.9) was associated with an increase in adhesion, when compared with a higher pH (7.2; P less than 0.05). Tween 20, Tween 80 and bovine serum albumin helped to eliminate a background of unbound bacteria from the test slides, but they also inhibited adhesion to some strains. Strain differences in adhesion and in ability to form a background were also observed. Some of these findings may have in vivo implications.     

Morphology of lacrimal gland in pig fetuses

The morphological and histological examinations of the lacrimal gland were conducted on pig fetuses coming from the 20th, 24th, 27th, 30th, 35th, 50th, 63rd, 94th and 112th day of gestation. The morphological examinations were carried out using the method of macroscopic preparation with a forehead magnifying glass and binocular (magnification 1.5-5.0x). In order to better visualize the anatomical elements, 60-80% absolute alcohol and 0.5-4% acetic acid solution were used for the examinations. On the 20th, 24th, 27th, and 30th day of gestation the whole fetuses were collected for the histological examinations. The whole eyeball with developing accessory organs was collected from the pig fetuses on the 35th day of gestation. On the 50th, 63rd, 94th and 112th day of gestation only the lacrimal gland was collected. Staining with H-E and Azan method was performed. On the 20th, 24th, 27th, 30th and 35th day of gestation ectodermal cells were not found in the collected material. On the 50th and 63rd day of gestation the connective tissue divides the gland parenchyma into indistinct lobes composed of gland cells. On the 94th day of gestation the number of lobes is substantially higher than on the 50th and 63rd day of gestation, while the number of lobules forming lobes decreases. On the 112th day of gestation each lobe is composed of 8-22 excretory ducts made up of the simple cuboid epithelium with a round nucleus arranged less or more peripherally.