Epidermal growth factor receptors and their ligands are expressed in the spleen of the Japanese Quail (Coturnix coturnix japonica) during the post-hatch period.

1. The epidermal growth factor (EGF) family plays an important role in the development, differentiation, migration and apoptosis of cells, as well as in wound healing, which are all essential to the viability of multicellular organisms. The avian spleen is a principal organ of systemic immunity and its importance in disease resistance is presumably accentuated by the scarcity of avian lymph nodes.2. The aim of this study was to determine whether EGF receptors (ErbB1-4) and their ligands (EGF, AREG and NRG) are expressed in the structural components of the quail spleen during the post-hatch period. At each selected age, from 1 d to 7, 14, 21 and 60 d, 10 quails were euthanised under ether anaesthesia and their spleens were fixed in a 10% formaldehyde-alcohol solution. Following routine histological processing, the streptavidin-biotin-peroxidase method was used for immunohistochemical examination.3. Strong cytoplasmic immunoreactions for ErbB2, ErbB4 and NRG were observed in the ellipsoid associated cells (EAC) of the quail spleen throughout the post-hatch period. This immunoreactivity in the EAC increased after the 7th d post-hatch. ErbB1 and ErbB3 immunoreactions were relatively similar and weak in all components of the spleen during the post-hatch period. Some immune cells of the peri-arterial lymphatic sheath (PALS) and peri-ellipsoidal lymphatic sheath (PELS) showed positive immunoreactivity for the ErbB receptors and their ligands. In the vascular smooth muscle cells, immunoreactivity for ErbB2 was stronger than that for the other ErbB receptors and their ligands.4. The data showed that ErbB receptors and their ligands (EGF, AREG and NRG) are expressed by different structural components of the quail spleen during the post-hatch period.

Expression of vascular endothelial growth factor receptors and their ligands in rat uterus during the postpartum involution period.

Vascular endothelial growth factor (VEGF) and its specific receptors, FLt1/fms, Flk1/KDR and FLt4, play important roles in vasculogenesis, and physiological and pathological angiogenesis. Whether angiogenic growth factors are involved in regulating angiogenic processes during the postpartum involution period (PP) of the rat uterus is unknown. We used immunohistochemistry to analyze the expression levels of VEGF, the fms-like tyrosine kinase 1 (FLt1/fms), the kinase insert domain-containing region 1 (Flk1/KDR), Fms-related tyrosine kinase 4 (FLt4) and vascular endothelial growth inhibitor (VEGI) in the rat uterus during the days 1, 3, 5, 10 and 15 of the PP to determine the temporal and spatial expressions of VEGF and its receptors during the PP. Throughout the PP, cytoplasmic and membrane staining of VEGI, VEGF and their receptors were observed in the lumens, crypts and glandular epithelial cells as well as in connective tissue and vascular endothelial and smooth muscle cells in the endometrium. We found that the intensity of the immunoreactions in the endometrium varied with the morphological changes that occurred during involution. Immunoreactions for VEGI, VEGF and their receptor, Flk1/KDR, in the luminal epithelial cells were stronger than those in the glandular epithelial and stromal cells, particularly during PP 1, 3 and 5, which suggests that these peptides may contribute to re-epithelialization of the endometrium. On the other hand, Flt1/fms immunoreactivity was strong mainly in the stromal cells during the PP. The presence of VEGF and its receptors (FLt1/fms, Flk1/KDR, FLt4) in the stromal cells and blood vessels during the PP suggests that they may contribute to regulating stromal repair and angiogenesis in the involuting uterus of the rat.

Endothelial and inducible nitric oxide synthase (NOS) immunoreactivity and NOS-associated NADPH-diaphorase histochemistry in the domestic cat (Felis catus) testis.

In this study, the cellular localization of nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) and the endothelial (eNOS) and inducible (iNOS) forms of nitric oxide (NO) synthase in the cat testis were studied using enzyme histochemical and immunohistochemical techniques. Stage-dependent nuclear and cytoplasmic eNOS/iNOS immunoreactivity and cytoplasmic NADPH-d reactivity were found in all germ cells, including spermatogonia, primary spermatocytes (preleptotene, zygotene, and pachytene spermatocytes), and round (Sa, Sb1) and elongating spermatids (Sb2, Sc) of the seminiferous epithelium. The pachytene spermatocytes exhibited strong positive reactions at all spermatogenic stage. Interestingly, in elongated spermatids (Sd1) at stages VI to VII, eNOS and iNOS immunostainings was observed only in the cytoplasm but not in the nuclei. eNOS and iNOS immunolabeling was observed in the acrosomal vesicle of some round spermatids (Sb1) at stages I, VII, and VIII, and in the acrosomal cap of elongating spermatids (Sb2) at stage II. Furthermore, eNOS, iNOS, and NADPH-d reactions in elongated spermatids (Sd2) just before spermiation at stage VIII were restricted only to the middle and principal pieces of the tail. Positive reactions were also observed in the Sertoli and Leydig cells as well as in other tissues including vascular endothelial and smooth muscle cells and peritubular myoid cells. These results suggest that NO may play an important role in chromatin condensation, spermatid shaping, and the final release of sperm from the spermatogenic epithelium. Furthermore, NO may also be involved in spermiogenesis, steroidogenesis, and apoptotic cell death.

Expression of survivin, Bcl-2 and Bax proteins in the domestic cat (Felis catus) endometrium during the oestrus cycle.

Apoptosis has been shown to be an important regulator of endometrium function. To clarify the regulation of apoptosis in the cat endometrium during the normal oestrus cycle, the expressions of the apoptosis-related proteins (Bcl-2 and Bax) and their correlation to the inhibitor of apoptosis protein Survivin were analysed using immunohistochemistry. The TUNEL technique (TdT-mediated dUTP nick end labelling) was also used to detect DNA fragmentation characteristic of apoptotic cells. The results demonstrated that TUNEL labelling is not effective for the detection of apoptosis in cat endometrium. Survivin was expressed in the luminal and glandular epithelial cells of cat endometrium during all phases of the oestrus cycle. Survivin was localized in both the cytoplasm and nuclei of superficial and deep uterine gland cells during the luteal phase, while only cytoplasmic staining was observed during the follicular and anoestrus phases. Bax immunoreactivity in the cytoplasm of luminal and glandular epithelial cells as well as the smooth muscle cells of blood vessels was weak in the anoestrus phase. Compared with anoestrus, the intensity of Bax immunostaining was moderate in the follicular phase and increased dramatically in the luteal phase. Bcl-2 immunostaining in the cytoplasm of luminal and glandular epithelial cells was moderate in the anoestrus phase. During the early follicular phase, cytoplasmic Bcl-2 immunostaining was detected mostly in glandular epithelial cells. In the mid-follicular phase, in glands, the amount of Bcl-2 protein increased progressively from the superficial to the deep layer. In contrast, the expression of Bcl-2 decreased in the secretory phase, being very low or absent in the mid- and late luteal phases. The overall results suggest that Survivin, Bax and Bcl-2 proteins may cooperatively contribute to cell apoptosis and cell proliferation in the cat uterus during the oestrus cycle.

Experimental hypothyroidism delays field excitatory post-synaptic potentials and disrupts hippocampal long-term potentiation in the dentate gyrus of hippocampal formation and Y-maze performance in adult rats.

Manipulations of thyroid hormones have been shown to influence learning and memory. Although a large body of literature is available on the effect of thyroid hormone deficiency on learning and memory functions during the developmental stage, electrophysiological and behavioural findings, particularly on propylthiouracil administration to adult normothyroid animals, are not satisfactory. The experiments in the present study were carried out on 12 adult male Wistar rats aged 6-7 months. Hypothyroidism was induced by administering 6-n-propyl-2-thiouracil in their drinking water for 21 days at a concentration of 0.05%. The spatial learning performance of hypothyroid and control rats was studied on a Y-maze. The rats were then placed in a stereotaxic frame under urethane anaesthesia. A bipolar tungsten electrode was used to stimulate the medial perforant path. A glass micropipette was inserted into the granule cell layer of the ipsilateral dentate gyrus to record field excitatory post-synaptic potentials. After a 15-min baseline recording of field potentials, long-term potentiation was induced by four sets of tetanic trains. The propylthiouracil-treated rats showed a significantly attenuated input-output (I/O) relationship when population spike (PS) amplitudes and field excitatory post-synaptic potentials (fEPSP) were compared. fEPSP and PS latencies were found to be longer in the hypothyroid group than in the control group. The PS amplitude and fEPSP slope potentiations in the hypothyroid rats were not statistically different from those in the control rats, except for the field EPSP slope measured in the post-tetanic and maintenance phases. The hypothyroid rats also showed lower thyroxine levels and poor performance in the spatial memory task. The present study provides in vivo evidence for the action of propylthiouracil leading to impaired synaptic plasticity, which might explain deficit in spatial memory tasks in adult hypothyroid rats.

The prenatal development and histochemistry of the ileal mucins in the bovine fetuses.

Few studies exist regarding the distribution of intestinal mucins in fetuses of mammalians such as cattle and sheep. In this study, we aimed to describe the changes in the mucin production by ileal epithelium of bovine fetuses during their prenatal development. The goblet cells showed heterogeneity in mucins and the apical cytoplasm of the enterocytes demonstrated Periodic acid Schiff-positive reaction which declined gradually towards the birth. Moreover, the number of the goblet cells containing acidic and mixed mucins augmented, whereas those containing neutral mucins decreased with advancing gestational age. After sixth month of gestation, with the initiation of the ileal Peyer patches and follicle-associated epithelium development, a gradual increase in the number of goblet cells containing sulfomucins was also noticed towards the birth. The presence of different mucins in the ileum of bovine fetuses throughout prenatal development might play a role in the protection of the intestinal mucosa against urinary waste products in swallowed amniotic fluid and bile. Furthermore, mucins can also contribute for the formation of meconium in intra-uterine life and building of strong intestinal barrier with predominating sulfomucins, protecting the intestine against potential pathogens and digestive enzymes after birth.

Structure of the oesophagus and morphometric, histochemical-immunohistochemical profiles of the oesophageal gland during the post-hatching period of Japanese quails (Coturnix coturnix japonica).

In the oesophagus, mucins, which originate from oesophageal submucosal glands, play an important role in the mucosal protection as a pre-epithelial barrier. In this study, the structure of cervical and thoracic parts of oesophagus of Japanese quail during the post-hatching period was compared, and the contents of carbohydrate and gastric mucin MUC5AC of the oesophageal glands in these parts were analysed at the light microscope levels by applying conventional histochemical and immunohistochemical methods. The oesophageal glands were present at hatching, located in the laminae propria. The numbers of glands were different in the cervical and thoracic parts, but the differences were found to be insignificant. The thoracic part has the oesophageal tonsils which are associated with the glands. Oesophageal tonsil was formed from day 5 after hatching. In quail of all ages, the secretory epithelium of glands contained neutral sialomucins and weakly sulphomucins. The cells in the neck region of secretory units contained sialomucins, while the cells of excretory ducts had strongly sulphomucins. Sialomucin containing cells in the secretory units increased with the advance of age and glandular development. But, in the secretory units, the sulphomucin content of glands was more in the thoracic part. The secretory epithelium of tonsil-associated glands contained mostly sulphomucins and a little sialomucin. From the hatching, MUC5AC mucin was detected in the cells of excretory ducts. Although the lymphoepithelium of the tonsil units exhibited negative reactions to all histochemical methods, it showed positive reaction to MUC5AC mucin antibody. In conclusion, the cervical and thoracic parts may be functionally different and the thoracic part of oesophagus was transformed into an immunological organ following day 5 after hatching.

Histochemical method for demonstrating quail mast cell types simultaneously.

The development and application of selective staining methods for routine detection of mast cells are of considerable interest, because these cells play an important role in health and disease. The composition of cytoplasmic mast cell granules depends on the species and type of mast cell. The study reported here was conducted to investigate the combined use of aldehyde fuchsin (AF) and the Alcian blue-critical electrolyte concentration (AB-CEC) (pH 5.8, 0.3 M MgCl(2)) techniques for differentiating avian mast cell subtypes. Tissue samples from skin, intestines, and lungs of six healthy adult quail and two control rats were fixed in Carnoy's solution and 10% formolin for routine histological processing. To determine the staining properties of sulfated glycosaminoglycans (GAGs), a three-step staining technique was applied using berberine sulfate, AF, and AB-CEC. In quail, AF positivity following application of the AB-CEC technique was found only in the lungs, mostly in cells that gave a berberine sulfate-positive reaction, and this positivity was determined to be localized particularly in the nucleus and perinuclear cytoplasm. In other regions, the pale AF staining of cells that did not emit fluorescence when stained with berberine sulfate was determined to be replaced by a blue color after application of AB-CEC. The AF/AB-CEC (pH 5.8, 0.3 M MgCl(2)) technique demonstrated that rat and quail mast cells varied in both GAG types and their distribution within the cell. Especially in avian species, this technique can be applied to distinguish mast cells according to their GAG content. It can be used as an alternative to the AB/safranin O staining procedure for differentiating mast cells that contain and lack heparin.

Immunolocalisation of serotonin, gastrin, somatostatin and glucagon in entero-endocrine cells of the goose (Anser anser).

The processes of digestion in the avian gastrointestinal tract depend on sophisticated control systems that co-ordinate secretion of digestive juices and movement of the luminal contents. In the current study, the distribution of serotonin-, gastrin-, glucagon- and somatostatin-immunoreactive endocrine cells was investigated by immunocytochemical methods in the intestinal tract of the goose. The number of cells immunoreactive for each antiserum was evaluated in different regions of the intestinal tract. Serotonin-, glucagon- and somatostatin-immunoreactive endocrine cells were seen throughout the intestinal tract, but somatostatin-immunoreactive cells were not detected in the colon of the goose. Gastrin-immunoreactive cells were detected only in the duodenum, jejunum and colon mucosa. It is concluded that the distribution pattern of the entero-endocrine cells in the goose is similar to that of most of the mammalian and other poultry species.

Histological and histochemical studies on the lingual, preglottal and laryngeal salivary glands of the Japanese quail (Coturnix coturnix japonica) at the post-hatching period.

The postnatal development and histochemistry of mucins of the lingual, preglottal and laryngeal glands in the quails were investigated by means of light microscopy using specific staining for complex carbohydrates. In this study, the tongues were taken from female and male quails from day 1 to day 60 after hatching. The salivary glands in quail's tongue comprised the lingual gland, with lateral and medial (paraentoglossal gland) portions that differ in morphology and histochemical staining, and the preglottal gland, with two lateral portions and one medial portion. The medial portion of the preglottal gland, which extended to the row of the laryngeal papillae on each side of the glottis, was described as the laryngeal gland. The salivary glands were present at hatching and their cells were functionally mature and secreted mucins. In quail of all ages, the histochemical reactions revealed that the cytoplasms of the secretory cells of the preglottal, laryngeal and paraentoglossal gland (medial portion of lingual gland) contained sialomucins and weakly sulphated epithelial mucins. Neutral mucins were absent in the paraentoglossal gland, while a small amount of neutral mucins was present in other glands. The mucins with vicinal diol groups, sialomucins and weakly sulphated epithelial mucins were mixed within the secretory cells of all the glands. All the histochemical reactions were restricted to the supranuclear regions of the secretory cells within the lateral portion of the lingual gland. In conclusion, the contents of mucins in the lingual, preglottal and laryngeal glands varied between different age groups, however, no differences in the glands' histochemistry between male and female quails were observed.

Quantitative histomorphology of liver growth in sheep at prenatal and postnatal stages.

This study was carried to determine quantitative histomorphologically on the development of the liver of sheep in prenatal and postnatal stages, and to prove the relationship between functional and structural differentiation of liver. There were more blood cells than hepatocytes, and haemotopoieisis was the primary function of the liver in the first half of gestation. As observed in the fetal stages bile ducts and Kiernan areas are formed from the 12th week. The distance between the two adjacent central veins was 401.2 = 20.8 micron in the fetuses and 629.77 -/+ 34.7 micron in the lambs, while rising in the adult to 740 + 14.35 micron. This increase was directly proportional to age. The average diameter of sheep hepatocyte and nuclei, and the ratio between the diameters of nuclei and their hepatic cells were compared according to the prenatal and postnatal stages, and the difference between these stages was found to be statistically significant (P < 0.05). This difference was sourced from adult sheep. The number of hepatocytes per unit area were 107.48 -/+ 6.63, 133.6 -/+ 7.01, 100.84 -/+ 6.63 in the fetus, lamb and adult liver of sheep, respectively, and the differentia that earned statistical importance was sourced from the young stages. The number of ductus biliferi was 2.75 -/+ 0.47 in the fetuses, however, this had risen to 5.8 -/+ 0.6 and 6.8 -/+ 0.37 respectively in the lambs and adult sheep. The portal lobule areas rose according to the age and were 0.17796 -/+ 0.00086 mm2 and 2.022650 -/+ 0.0097 mm2 respectively in the lambs and adult sheep and the differentia between young and adult sheep was statistically important (P < 0.05).

[The bacteriocin typing of Klebsiella pneumoniae strains]. / Bakteriotsinotipirovanie shtammov Klebsiella pneumoniae.

Results of bacteriocin typing of 196 strains of the Klebsiella genus are presented. They are typified by their sensitivity to bacteriocins and by their production using colicinogenic and indicating strains from collection of P. Fredericq [correction of Frederick], D. G. Kudlai, N. I. Koshanova as well as klebocinogenic K-type cultures of Klebsiella previously suggested by the authors. Investigation results have shown sufficient stability of a bacteriocinotype of the cultures confirmed by the population analysis. It is concluded that bacteriocin typing may be recommended as an additional method in epidemiological labelling of Klebsiella cultures.