Expression patterns of intestinal calcium transport factors and ex-vivo absorption of calcium in horses

BACKGROUND: In many species, the small intestine is the major site of calcium (Ca(2+)) absorption. The horse differs considerably from most other species with regard to the physiology of its Ca(2+) metabolism and digestion. Thus, this study was performed to get more information about the transcellular Ca(2+) absorption in the horse.Two mechanisms of intestinal Ca(2+) absorption are described: the passive paracellular pathway and the active, vitamin D-dependent transcellular pathway. The latter involves the following elements: vitamin D receptors (VDR), transient receptor potential vanilloid channel members 5 and 6 (TRPV5/6), calbindin-D9k (CB), the Na/Ca exchanger (NCX1) and the plasma membrane Ca-ATPase (PMCA). The aim of the present study was to investigate the protein and mRNA expression patterns of VDR, CB and TRPV6 and the ex-vivo Ca(2+) absorption in horses, assessed by qualitative and quantitative RT-PCR, western blot, immunohistochemistry and the Ussing chamber technique. RESULTS: Highest CB and TRPV6 mRNA levels were detected in the duodenum as compared to the middle parts of the jejunum and ileum and several sites of the large intestine. VDR mRNA levels did not change significantly throughout the intestine. TRPV5 mRNA was not detectable in the horse intestine. The highest VDR and CB protein levels were measured in the duodenum. Ussing chamber studies revealed ex-vivo Ca(2+) absorption only in the duodenum, but not in cecum and specific sites of the colon. CONCLUSION: The present findings suggest that TRPV6, CB and VDR may be involved in active intestinal Ca(2+) absorption in horses, as described for other mammals. TRPV5 may not play a major role in this process. Furthermore, the expression patterns of these Ca(2+) transport elements and the results of the Ussing chamber procedure indicate that a significant part of active intestinal Ca(2+) absorption occurs in the duodenum in this species

An animal model in sheep for biocompatibility testing of biomaterials in cancellous bones

BACKGROUND: The past years have seen the development of many synthetic bone replacements. To test their biocompatibility and ability for osseointegration, osseoinduction and -conduction requires their placement within bone preferably in an animal experiment of a higher species. METHODS: A suitable experimental animal model in sheep with drill holes of 8 mm diameter and 13 mm depth within the proximal and distal humerus and femur for testing biocompatibility issues is introduced. RESULTS: This present sheep model allows the placing of up to 8 different test materials within one animal and because of the standardization of the bone defect, routine evaluation by means of histomorphometry is easily conducted. This method was used successfully in 66 White Alpine Sheep. When the drill holes were correctly placed no complications such as spontaneous fractures were encountered. CONCLUSION: This experimental animal model serves an excellent basis for testing the biocompatibility of novel biomaterials to be used as bone replacement or new bone formation enhancing materials

Placental Origin of Prostaglandin F 2 α

In the present study, the question was addressed whether the feline placenta can synthesize prostaglandin F2 α (PGF2 α ). The PGFS protein was elevated, particularly at 2.5-3 weeks of pregnancy compared to 7-8 (P < 0.05) and 8.5-9 weeks (P < 0.001). Transcripts for PGFS were significantly upregulated at 2.5-3 weeks of pregnancy and then gradually declined towards the end of gestation (P < 0.001). Transcripts for PTGS2 were only upregulated in placentas from queens close to term (P < 0.001) compared with earlier phases. Staining of PTGS2 showed distinct positive signals in placentas obtained during the last week before labor, particularly in the strongly invading trophoblast surrounding blood vessels, and also in decidual cells. Shortly after implantation, signals for PGFS were localized in the trophoblast cells. Near term, PGFS staining was seen mainly in decidual cells. Both placental PGF2 α and plasma PGFM were elevated towards the end of pregnancy (P < 0.001) compared with earlier weeks of pregnancy. The content of PGF2 α in extracted placenta mirrored the PGFM level in plasma of pregnant females. During late gestation there is a significant increase in PGFM levels in maternal blood and of PGF2 α levels in placental tissue concomitant with an upregulation of placental PTGS2

Luteal and placental function in the bitch: spatio-temporal changes in prolactin receptor (PRLr) expression at dioestrus, pregnancy and normal and induced parturition

Background: Endocrine mechanisms governing canine reproductive function remain still obscure. Progesterone (P4) of luteal origin is required for maintenance of pregnancy. Corpora lutea (CL) are gonadotrop-independent during the first third of dioestrus; afterwards prolactin (PRL) is the primary luteotropic factor. Interestingly, the increasing PRL levels are accompanied by decreasing P4 concentrations, thus luteal regression/luteolysis occurs in spite of an increased availability of gonadotropic support. PRL acts through its receptor (PRLr), the expression of which has not yet been thoroughly investigated at the molecular and cellular level in the dog. Methods: The expression of PRLr was assessed in CL of non-pregnant dogs during the course of dioestrus (days 5, 15, 25, 35, 45, 65 post ovulation; p.o.) as well as in CL, the utero/placental compartments (Ut/Pl) and interplacental free polar zones (interplacental sites) from pregnant dogs during the pre-implantation, post-implantation and mid-gestation period of pregnancy and during the normal and antigestagen-induced luteolysis. Expression of PRLr was tested by Real Time PCR, immunohistochemistry and in situ hybridization. Results: In non-pregnant CL the PRLr expression was significantly upregulated at day 15 p.o. and decreased significantly afterwards, towards the end of dioestrus. CL of pregnancy showed elevated PRLr expression until mid gestation while prepartal downregulation was observed. Interestingly, placental but not interplacental expression of PRLr was strongly time-related; a significant upregulation was observed towards mid-gestation. Within the CL PRLr was localized to the luteal cells; in the Ut/Pl it was localized to the fetal trophoblast and epithelial cells of glandular chambers. Moreover, in mid-pregnant animals treated with an antigestagen, both the luteal and placental, but not the uterine PRLr were significantly downregulated. Conclusions: The data presented suggest that the luteal provision of P4 in both pregnant and non-pregnant dogs may be regulated at the PRLr level. Furthermore, a role of PRL not only in maintaining the canine CL function but also in regulating the placental function is strongly suggested. A possible functional interrelationship between luteal P4 and placental and luteal PRLr expression also with respect to the prepartal luteolysis is implied

Sistem pengendalian internal dalam upaya mencegah penyimpangan dana pembiayaan di KSPPS BMT Aulia Magelang

Pengendalian internal merupakan sistem dan prosedur yang digunakan perusahaan untuk mencapai sasaran dan tujuan yang diinginkan, yaitu menghasilkan laporan keuangan yang akurat serta mendorong ketaatan terhadap kebiajakn dan peraturan yang telah ditetapkan. sistem pengendalian mencegah penyimpangan dana pembiayaan pada KSPPS Amanah Usaha Mulia Magelang dapat diambil sebagai berikut : Sistem Pengendalian Internal KSPPS Amanah Usaha Mulia Magelang. Sistem pengendalian internal yang digunakan KSPPS Amanah Usaha Mulia Magelang dalam pencegah penyimpangan dana pembiayaan yaitu dengan melakukan pengecekan rutin untuk mencegah resiko yang tidak diinginkan. setiap pegawai dituntut untuk jujur dan bertanggungjawab dengan tugas dan wewenangnya masing-masing, dan paling penting semua pegawai saling berkerjasama untuk mencapai tujuan yang telah ditetapkan KSPPS Amanah Usaha Mulia Magelang. Efektivitas sistem pengendalian internal KSPPS Amanah Usaha Mulia Magelang. Keefektivitasan sistem pengendalian internal KSPPS Amanah Usaha Mulia Magelang sudah berjalan sesuai dengan perencanaan yang telah dituju, dalam pengoperasiannya juga sudah sesuai dengan prinsip dan komponen di pengendalian internal. Prinsip dan komponen tersebut antara lain : pegawai yang berkualitas dan dapat dipercaya, pemisahan wewennag, pengawasan, penetapan tanggungjawab secara perseorangan, pencatatan yang seksama dengan segera, penjagaan fisik, pemisaan oleh petugas yang bebas dari tugas rutin merupakan prinsip pengendalian internal dan komponen pengendalian meliputi lingkungan pengendalian, penentuan risiko manajemen, aktivitas pengendalian, informasi dan komunikasi dan pemantauan

Endocrine control of canine mammary neoplasms: serum reproductive hormone levels and tissue expression of steroid hormone, prolactin and growth hormone receptors

BACKGROUND: Neoplasms of the mammary gland are among the most common diseases in female domestic dogs (Canis familiaris). It is assumed that reproductive hormones influence tumorigenesis in this species, although the precise role of the endocrine milieu and reproductive state is subject to continuing discussion. In line with this, a recent systematic review of available data on the development of mammary neoplasms revealed weak evidence for risk reduction after neutering and an effect of age at neutering. Investigation of several hormone receptors has revealed decreased expression of estrogen receptor-alpha (ERα, ESR1), progesterone (P4) receptor (PGR), prolactin (PRL) receptor (PRLR) and growth hormone receptor (GHR) associated with neoplastic differentiation of mammary tissues. In other studies, increased levels of estrogens, progesterone and prolactin were found in serum and/or tissue homogenates of dogs with malignant neoplasms. However, the association between these entities within one animal population was never previously examined. Therefore, this study investigated the association between circulating serum concentrations of estradiol-17β, progesterone and prolactin, and gene expression of ERα (ESR1), ERβ (ESR2), PGR, PRLR, PRL and GHR, with respect to reproductive state (spayed vs. intact) and cycle stage (anestrus vs. diestrus). Additionally, the expression of E-cadherin (CDH-1) was evaluated as a possible indicator of metastatic potential. RESULTS: For all receptors, the lowest gene expression was found in malignant tumors compared to normal tissues of affected dogs. Steroid levels were not influenced by their corresponding receptor expression in mammary neoplasms, but increased PRL levels were negatively associated with low PRLR gene expression in malignant tumors. The expression of CDH-1 was influenced by tumor malignancy and cycle stage, i.e., the highest gene expression was found in benign mammary tumors in diestrous dogs compared to normal and malignant mammary tissues of anestrous and spayed dogs. CONCLUSIONS: Herein, it has been confirmed that transformation towards malignant neoplasms is associated with significant reduction of gene expression of particular hormone receptors. Only PRLR in malignant tumors seems to be influenced by circulating PRL levels. In dogs, CDH-1 can be used as a prognostic factor; its expression, however, in benign tumors is influenced by cycle stage

Influence of feeding and UVB exposition on the absorption mechanisms of calcium in the gastrointestinal tract of veiled chameleons (Chamaeleo calyptratus)

The purpose of this study was to investigate the influence of feeding and UVB exposition on the occurrence and distribution patterns of vitamin D receptors (VDR) and calbindin D28k (Cb-D28k) in the gastrointestinal tract of veiled chameleons. Thus, 56 veiled chameleon hatchlings were divided into six treatment groups: UV (with UVB exposure); No (no supplements, no UVB exposure); CaAUV (with calcium (Ca), vitamin A supplementation, UVB exposure); CaA (with Ca, vitamin A supplementation); CaADUV (with Ca, vitamin A, vitamin D supplementation, UVB exposure); and CaAD (with Ca, vitamin A, vitamin D supplementation). Animals were reared under the suspected conditions for 6 months on locust-based diets. Tissue samples of stomach, duodenum, ileum and colon were taken, and semi-quantitative immunohistochemical methods (IHC) were performed to detect Cb-D28k and VDR. VDR immunoreactions were higher in the luminal epithelium of the duodenum than in that of the ileum. VDR immunoreactions in the luminal epithelium were higher at the base of the villi of the duodenum as compared to the tip. Cb-D28k immunoreactions were mainly observed in the luminal epithelium of the duodenum. The two groups treated with all dietary supplements (CaADUV, CaAD) exhibited a higher Cb-D28k immunoreaction as those with no supplements and UVB exposure only. No immunoreaction for both proteins could be detected in the stomach. This study suggests that the duodenum plays an important role in the active transcellular absorption of Ca in veiled chameleons as shown by the immunohistochemical detection of VDR and Cb-D28k. Expression of Cb-D28k, in particular, appears to be regulated by dietary supplementation of vitamin D and vitamin A. VDRs, however, tended to be upregulated when animals were not supplemented with Ca, vitamin D and vitamin A. This may be due to the decreased Ca concentrations which caused vitamin D activation in the skin without any supplementation, but UVB exposure

Expression of GnRH receptor in the canine corpus luteum, and luteal function following deslorelin acetate-induced puberty delay

The goals of this study were as follows: (Experiment 1) to examine the basic capability of canine corpora lutea (CL) to respond to GnRH by assessing expression of gonadotropin-releasing hormone receptor (GnRH-R) in luteal samples collected throughout the luteal lifespan from non-pregnant dogs, and (Experiment 2) to investigate the effects of pre-pubertal application of the GnRH agonist deslorelin acetate on luteal function following the first oestrus. Mature CL were collected during the mid-luteal phase (days 30–45) from treated and control bitches. Transcript levels of several factors were determined: estrogen receptors (ESR1/ERα, ESR2/ERβ), progesterone (P4)-receptor (PGR), prolactin receptor (PRLR), PGE2-synthase (PTGES) and PGE2 receptors (PTGER2/EP2, PTGER4/EP4), vascular endothelial growth factor (VEGFA) and VEGF receptors (VEGFR1 and VEGFR2), cyclooxygenase 2 (COX2/PTGS2), steroidogenic acute regulatory protein (STAR) and 3β-hydroxysteroid dehydrogenase (3βHSD). Additionally, levels of Kisspeptin 1 (Kiss1) and its receptor (KISS1-R) were evaluated. Although generally low, GnRH-R expression was time dependent and was elevated during early dioestrus, with a significant decrease towards luteal regression. In deslorelin-treated and control dogs, its expression was either low or frequently below the detection limit. EP2 and VEGFR1 were higher in the treated group, which could be caused by a feedback mechanism after long-term suppression of reproductive activity. Despite large individual variations, 3βHSD was higher in the deslorelin-treated group. This, along with unchanged STAR expression, was apparently not mirrored in increased luteal functionality, because similar P4 levels were detected in both groups. Finally, the deslorelin-mediated long-term delay of puberty does not have negative carry-over effects on subsequent ovarian functionality in bitches

Centrosome clustering in the development of bovine binucleate trophoblast giant cells

Binucleate trophoblast giant cells (BNC) are the characteristic feature of the ruminant placenta. During their development, BNC pass through 2 acytokinetic mitoses and become binucleate with 2 tetraploid nuclei. In this study, we investigate the number and location of centrosomes in bovine BNC. Centrosomes typically consist of 2 centrioles surrounded by electron-dense pericentriolar material. Duplication of centrosomes is tightly linked to the cell cycle, which ensures that the number of centrosomes remains constant in proliferating diploid cells. Alterations of the cell cycle, which affect the number of chromosome sets, also affect the number of centrosomes. In this study, we use placentomal tissue from pregnant cows (gestational days 80-230) for immunohistochemical staining of γ-tubulin (n = 3) and transmission electron microscopy (n = 3). We show that mature BNC have 4 centrosomes with 8 centrioles, clustered in the angle between the 2 cell nuclei. During the second acytokinetic mitosis, the centrosomes must be clustered to form the poles of a bipolar spindle. In rare cases, centrosome clustering fails and tripolar mitosis leads to the formation of trinucleate "BNC". Generally, centrosome clustering occurs in polyploid tumor cells, which have an increased number of centrioles, but it is absent in proliferating diploid cells. Thus, inhibition of centrosome clustering in tumor cells is a novel promising strategy for cancer treatment. BNC are a cell population in which centrosome clustering occurs as part of the normal life history. Thus, they might be a good model for the study of the molecular mechanisms of centrosome clustering