Regulatory expression of Brachyury and Goosecoid in P19 embryonal carcinoma cells.

Mouse P19 embryonal carcinoma cells can differentiate into various cell types depending on culture conditions. Here we show that the expression of the mesodermal genes Brachyury (Bra) and Goosecoid (Gsc) are under regulatory control in P19 cells. When P19 cells were cultured in a tissue culture dish in the presence of serum, Bra and Gsc were unexpectedly expressed.

Regulation of melanin synthesis by the TGF-beta family in B16 melanoma cells.

Effects of representative members of the transforming growth factor-beta (TGF-beta) family, TGF-beta1, activin A and BMP-2, on melanin content and expression of pigment-producing enzymes were examined in B16 melanoma cells. Treatment with TGF-beta1 or activin A but not with BMP-2 significantly decreased melanin content and expression of Tyrosinase and Tyrp-1, suggesting an inhibitory effect of TGF-beta1 and activin A on melanin synthesis. TGF-beta1 completely inhibited melanin synthesis induced by alpha-melanin stimulating hormone (alpha-MSH), whereas activin A only slightly did.

Expression and function of alternative splice variants of the mouse TGF-beta type I receptor.

Mouse TGF-beta type I receptor, Alk5, encodes two types of mRNA generated by alternative splicing of four amino acids in the extracellular region. In the present study, the expression and function of alternative splice variants of Alk5, i.e.

A macroscopic examination of M. biceps femoris and M. gluteus maximus in the orangutan.

The musculature of the hip and thigh in the orangutan has been described previously. Anatomically, there are various descriptions among primates in those structures, in particular, the relationship between M. biceps femoris and M.

Factors affecting struvite (MgNH4PO4.6H2O) crystallization in feline urine.

Factors affecting struvite, a magnesium-ammonium-phosphate complex (MgNH(4)PO(4).6H(2)O), in feline urine were evaluated. Incubation of just "urine mineral (UM)" solution, in which mineral concentrations are compatible with those in feline urine, for 4 h at 37 degrees C did not induce the formation of crystals.

A sensitive detection of phospho-Smad1/5/8 and Smad2 in Western blot analyses.

The transforming growth factor-beta (TGF-beta) family is involved in a variety of physiological processes, and transmits signals through phosphorylation of Smad by the receptor complexes. In the present study, effects of blocking solution in Western blot analyses on detection of phosphorylated Smad1/5/8 and Smad2 were examined. When EzBlock was used as a blocking reagent, phosphorylated Smad1/8 and Smad2 were most efficiently detected.

Gene expression of secretory carbonic anhydrase isozymes in striated ducts of canine salivary glands using laser microdissection system.

To clarify whether striated duct cells in canine salivary glands synthesize secretory carbonic anhydrase (CA-VI), as is the case with serous acinar cells, the present study utilized laser microdissection to harvest striated duct cells from canine parotid and submandibular glands, and total RNA extracted from these cells was then amplified by reverse transcription-polymerase chain reaction to assess CA-VI gene expression. The results confirmed the local expression of CA-VI mRNA in striated duct cells. This clarified that, in canine salivary glands, CA-VI is synthesized in not only serous acinar cells, but also striated duct cells.

Spermatogenesis in aged rats after prenatal 3,3',4,4',5-pentachlorobiphenyl exposure.

We previously reported that prenatal exposure to 3,3',4,4',5-pentachlorobiphenyl (PCB126) had dose-related adverse effects on the spermatogenesis of 7(pubescent)- and 17(adult)-week-old rats, but the effects in middle and old age have been unclear. In this study, the spermatogenesis of male Sprague-Dawley rats whose dams had been injected (i.g.

The evaluation of autonomic nervous system activation during learning in rhesus macaques with the analysis of the heart rate variability.

The aims of this study were to measure the activity of the autonomic nervous system using heart rate variability (HRV) during learning tasks and to clarify the relationship between learning to overcome a difficult situation and the autonomic nervous system in monkeys. Two young male monkeys (Macaca mulatta) were given simple discrimination learning tasks (DL) and delayed matching to samples tasks (DMTS); Holter-type electrocardiography was done, and HRV was measured. We defined the frequency bands of HRV in rhesus macaques; the low frequency (LF) was 0.

Prenatal exposure to 3,3',4,4',5-pentachlorobiphenyl (PCB126) promotes anxiogenic behavior in rats.

Polychlorinated biphenyls (PCBs) are environmental contaminants that have adverse effects on the endocrine and nervous systems. As they are still detected in breast milk and adipose tissue in humans, the accumulated PCBs may transfer from mothers to children and damage central nervous system. It is revealed from epidemiological studies that cognitive and motor functions were damaged in children born to mothers who ingested PCBs-contaminated foods.

Expression and transcriptional activity of alternative splice variants of Mitf exon 6.

Microphthalmia-associated transcription factor (Mitf) is a tissue-specific transcription factor. At least nine distinct mouse isoform mRNAs are encoded by alternative splicing of the first exon of Mitf (Mitf-A, -B, -C, -D, -E, -H, -J, -M, and -mc), while exons 2-9 of all Mitf isoforms examined to date are identical. In addition, alternative splice variants of exon 6a encoding 6 amino acid proximal to the basic region of the protein are known in Mitf-A, -H, and -M.

Increased number of whistles of bottlenose dolphins, Tursiops truncatus, arising from interaction with people.

The acoustic mode is the most reasonable means for social animals such as dolphins to maintain contact in the underwater habitat, and has been developed since they moved to the sea. This study investigates variations in dolphin vocalizations under the following conditions in a captive environment: 1) before feeding (Pre-feeding), 2) during feeding (Feeding), 3) during free time without the presence of people (Free), 4) during interaction with people located upon a float (Float), 5) during interaction with people in the water (Water). During the experiments, a total of 2642 whistles were extracted from sonogram data using a spectrogram.

The time course of lymphatic routes emanating from the peritoneal cavity in rats.

The lymph drainage routes from the abdominal cavity in rats were observed at 3 min, 1, 2 and 4 h after India ink was administered intraperitoneally. Four systems of lymph drainage routes from the peritoneal cavity were observed. Three minutes after injection, the drainage route travelled via the intrathoracic lymph vessels located along the internal thoracic artery and returned to the anterior mediastinal lymph nodes.

Immunohistolocalization and gene expression of the secretory carbonic anhydrase isozymes (CA-VI) in canine oral mucosa, salivary glands and oesophagus.

The immunohistolocalization of secretory carbonic anhydrase isoenzymes (CA-VI) in canine salivary glands, parotid, submandibular, sublingual and zygomatic glands, oral and oesophageal mucosa was studied using a specific antiserum against a canine CA-VI. In addition, the gene expression of CA-VI from the same tissue was studied using a real-time reverse-transcriptase polymerase chain reaction. In all salivary glands and oesophageal gland, immunostaining intensely localized CA-VI antiserum throughout the cytoplasm of serous acinar cells, including serous demilune and ductal epithelial cells.

A sensitive gait parameter for quantification of arthritis in rats.

Quantification of arthritis is helpful for investigating pain mechanisms of arthritis and for developing new drugs. We assessed and identified a feasible parameter for quantification of rat arthritis using a novel gait analyzing system. Knee-joint injection with small doses of lambda-carrageenan decreased swing time ratio (STR, swing time of the non-treated hindlimb/swing time of the lambda-carrageenan-injected hindlimb) in a dose-dependent manner.

Plasma concentration of vitamin C in dogs with a portosystemic shunt.

Most mammals, including dogs, synthesize vitamin C in the liver. We measured the plasma concentration of vitamin C to assess the body vitamin C status in 15 dogs with a portosystemic shunt (PSS). The plasma biochemical parameters indicated liver abnormalities in all the dogs.

Localization of Ang-1, -2, Tie-2, and VEGF expression at endothelial-pericyte interdigitation in rat angiogenesis.

Endothelial cells and pericytes play critical role in angiogenesis, which is controlled, in part, by the angiopoietin (Ang)/Tie-2 system and vascular endothelial growth factor (VEGF). Here, we investigated Ang, Tie-2, and VEGF expression within endothelial cells and pericyte interdigitations (EPI), which consist of cytoplasmic projections of pericytes and corresponding endothelial indentations. After subcutaneous implantation of a thermoreversible gelation polymer disc in rats, the capillary density was low on day 5, increased to a peak on day 7, and then decreased on days 10-20.

Parathyroid hormone-related protein (PTHrP) and Ca levels in the milk of lactating cows.

To investigate the roles of mammary PTHrP in calcium uptake and/or release in the mammary gland of cows, plasma PTHrP and Ca levels, and their arterial-venous differences were examined in a Jersey cow during the periparturient period. Levels of Ca in both abdominal aorta and abdominal subcutaneous vein blood slightly decreased around the parturition and at 24 days after the parturition, however, no remarkable arterial-venous differences were observed. Plasma PTHrP levels in both arterial and venous samples were below the detection limit (0.

Identification of tocopherol-associated protein as an activin/TGF-beta-inducible gene in mast cells.

Previous studies have demonstrated that treatment with activin A and TGF-beta(1), members of the TGF-beta family, stimulated maturation of mouse bone marrow-derived cultured mast cells (BMMC), which was characterized by morphology and gene expression of mouse mast cell proteases (mmcps). In order to gain a better understanding of activin A- and TGF-beta(1)-induced maturation in mast cells, we investigated the genes that were up-regulated in response to treatment with these two members of the TGF-beta family. The cDNA microarray analyses indicated that in BMMC, five genes were induced by treatment with 4 nM activin A for 2 h.

Requirement of Smad3 for mast cell growth.

The involvement of the TGF-beta family in cell growth of bone marrow-derived mast cells (BMMC) cultured with medium containing pokeweed mitogen-stimulated spleen cell-conditioned medium (PWM-SCM) was examined. Doubling time of BMMC from Smad3-null mice was longer than that from wild-type (WT) mice, and the differences tended to be larger with time of culture. Consistent with the results, uptake and reduction of [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS] was lower in Smad3-deficient BMMC.

Expression of bovine parathyroid hormone/parathyroid hormone- related protein (PTH/PTHrP) receptor mRNA in the mammary gland of cows.

To examine the PTH/PTHrP receptor in the mammary gland, molecular cloning of bovine PTH/PTHrP receptor and measurement of its mRNA expression were carried out in cows during the periparturient period. The PTH/PTHrP receptor gene was partially cloned, and expression of bovine PTH/PTHrP receptor mRNA was detected in various tissues of the cow. In the mammary gland, PTH/PTHrP receptor mRNA expression was constantly low during the periparturient period, whereas PTHrP mRNA expression dramatically increased after parturition.

Involvement of p38 MAP kinase and Smad3 in TGF-beta-mediated mast cell functions.

Transforming growth factor-beta (TGF-beta) modulates functions of bone marrow-derived cultured mast cells (BMMCs); cell maturation (up-regulation of mouse mast cell proteases (mmcps)), growth arrest and migration. We investigated the roles of p38 MAP kinase and Smad3 in TGF-beta-mediated cell responses in BMMCs. Treating BMMCs with TGF-beta induced the phosphorylation of p38 within 2 h and persisted for 24 h.

Transcriptional regulation of mouse mast cell protease-7 by TGF-beta.

Mouse mast cell protease-7 (mmcp-7) is a tryptase predominantly expressed in differentiated connective tissue-type mast cells. Previous study revealed that transforming growth factor-beta (TGF-beta) increases gene transcript of mmcp-7 in mast cells. The present study explored molecular mechanism of the up-regulation of mmcp-7 by TGF-beta.

Evaluation of meat meal, chicken meal, and corn gluten meal as dietary sources of protein in dry cat food.

The nutritional value of meat meal (MM), chicken meal (CM), and corn gluten meal (CGM) as dietary sources of protein in dry food formulated for adult cats was evaluated. Twelve healthy adult cats (11 males and 1 female) were used. Dry diets containing MM, CM, or CGM as the main protein source were given for a 3-week period in a 3 x 3 Latin-square design.

Responses during cell preparation for functional analyses in mouse bone marrow-derived cultured mast cells.

Murine bone marrow-derived cultured mast cells (BMMCs) are most widely used in in vitro experiments for evaluation of mast cell functions. The present study has shown that cell preparation procedure, i.e.