The notion of how malnutrition early in life affects ontogenesis has evolved considerably since the mid-1960s. Since then, there have been many studies on the effects of early malnutrition. Nutritional and metabolic exposure during critical periods in early human and animal development may have long-term programming effects in adulthood. This is supported by evidence from epidemiological studies, numerous animal models and clinical intervention trials. In this paper, we review the effects of early malnutrition on cognitive function, metabolic syndrome, immunity and the gastrointestinal tract, as well as possible underlying mechanisms, and consider diarrhoeal disease and poor cognitive function as examples for understanding the interrelation of the harmful effects caused by early malnutrition. Previous studies on early malnutrition have mainly concentrated on humans and rats. Therefore, the main aim of the present review was to give animal scientists a clear understanding of the harmful effects of early malnutrition on animal growth and animal production, and to help identify appropriate feeding techniques to prevent early malnutrition.

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The caspases (Casps) are a family of cysteine proteases that are known to regulate apoptotic signaling. Apoptosis by activation of Casp is strongly associated with embryonal development and regeneration in many organs, therefore indicating that disorders caused by homozygous mutation in Casp genes can result in embryonic lethality. In the present study, the authors investigated the causative relationship between skeletal myogenesis and the activation of Casps by analyzing their dynamics during mouse embryogenesis. Individual myogenetic tissues were obtained from C57BL/6 mouse embryos aged 12.5-17.5 days post-conception (dpc), and the expression of Casps was analyzed by histochemical and molecular biological methods. Immunoreactions for Casp-3, -9 and -12 were detected first in myoblasts, increasing according to embryonal development, as a result of which myoblasts differentiated into myotube cells. On the other hand, the immunoreaction for ssDNA, which is well-known as an apoptosis marker, was little detected during the skeletal myogenesis. Quantification analysis for Casp mRNA expression by RT-PCR as well as by in situ hybridization showed a peak at 15.5 dpc but a decrease at 17.5 dpc. Similar dynamics were detected for Myod1 mRNA, one of the muscle regulatory factors, but not for Fasl, Bax and Rock1, apoptosis-associated factors during skeletal myogenesis. These results suggest that the activation of Casps in skeletal myogenesis is deeply associated with myoblast differentiation, but not directly related to apoptosis.

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The fine structure in the center and periphery of the cornea of 16 beagle dogs were characterized and compared. The central cornea (about 540 μm) was apparently thinner than the peripheral cornea (about 720 μm). Thickness ratios of the corneal substantia propria to the entire cornea were approximately 86% in both portions. In addition, number of collagen lamellae, collagen fibril diameter, and collagen fibril index of the central substantia propria are different from those of the periphery (253 vs 236 lamellae, 29.1 vs 32.0 nm, and 39.0 vs 41.6%, respectively). These differences are thought to be due to site-dependent accumulation of proteoglycans (decorin and lumican) which are responsible for production of thin fibrils. The central portion with higher proteoglycans would have abundant thin fibrils with less slippage but better elasticity to buffer against the direct impact of intraocular pressure on the cornea. In contrast, thick fibrils in the peripheral substantia propria would contribute to the maintenance of tensile strength acting on the transition zone between the cornea and sclera.

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Sexual differences in Japanese Jungle Crows (Corvus macrorhynchos) are not obvious because both sexes have black plumage. However, closer examination reveals that their plumage color is not only black; it is also iridescent color. Furthermore, the iridescence is more pronounced in adult males than that in females. The iridescence seems to be related to the density and concentration of melanin granules. The density of melanin granule and the concentration of eumelanin in feathers were significantly higher in males than that in females (P<0.01). However, the amount of pheomelanin was below the detection limits in this study.

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Polymerase chain reaction (PCR) coupled with direct sequencing of the product of the hexon gene was applied to avian adenoviruses (formerly group I-III). The expected sizes of DNA fragments were successfully amplified by PCR from all of the group I-III avian adenoviruses with our designed primers. The resulting PCR product contained diagnostically relevant hexon sequences that could be used to identify the group and type of avian adenovirus.

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The presence of 11 virulence-associated genes in Campylobacter jejuni isolated from chickens at different stages of their development was examined by PCR. Multiple groups of C. jejuni were colonized in one chicken at the same time. Two hundred thirty one C. jejuni in total were isolated from the same group at four different ages and 12 groups of C. jejuni possessing of 11 virilence-associated genes were observed. Eleven, eight, five and three groups of C. jejuni were detected at 21, 28, 42 and 56 days after hatching, respectively. The variation of Campylobacter groups was reduced as the chicken developed and one group of C. jejuni became predominant.

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The serum cystatin C (Cys-C) concentration is a better filtration marker than plasma creatinine (Cre) concentration in humans. In veterinary medicine, a few studies have shown that the serum Cys-C concentration in dogs is also a better marker than the plasma Cre concentration. The purpose of this study is to evaluate the applicability of measuring the serum Cys-C concentration by an enzyme-linked immunosorbent assay (ELISA) as a marker of the glomerular filtration rate in dogs with various renal dysfunctions. The serum Cys-C concentration in dogs with chronic kidney disease (CKD) was significantly higher (1.23 ± 0.21 mg/L) than that in 76 control dogs (0.85 ± 0.15) (P<0.001). The reference range of the serum Cys-C concentrations in samples from the 76 control dogs was 0.55-1.15 mg/l. Serum Cys-C concentration was more strongly correlated with plasma iohexol clearance (r=-0.704, P<0.001) than plasma Cre concentration in dogs (r=-0.598, P<0.001). In a receiver operating characteristics analysis, significant differences between the serum Cys-C and plasma Cre concentrations were found with regard to their AUC (0.949, [SE, 0.019] and 0.849 [SE, 0.029]) and diagnostic sensitivity (90.3% and 73.6%) for detecting decreased PCio (P<0.05). Therefore, the measurement of serum Cys-C concentration by ELISA is more useful for the detection of early CKD than measuring the plasma Cre concentration.

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The immunopathology behind atopic dermatitis (AD) involves a myriad of inflammatory cells and their mediators. Thymus and activation-regulated chemokine (TARC) plays a significant role in the inflammatory phase by recruiting CCR4⁺ T_H2 cells. In addition, CD25⁺ activated T cells further propagate the allergic response after sensitization by producing cytokines. The purpose of this pilot study was to evaluate how exposure to a common allergen (house dust mite, HDM) would affect the proportions of circulating CD4⁺CCR4⁺ T_H2 cells and CD4⁺CD25⁺activated T cells in an experimental model of AD using high-IgE Beagles. In this experimental model, previously sensitized Beagles develop lesions and pruritus upon allergen challenge consisting of 3-day environmental exposure, 3 hours/day. Blood samples were obtained before, during, and after the end of challenge (days 0, 2, 4, and 17). Clinical signs were evaluated and scored at the same time points. Peripheral blood mononuclear cells (PBMCs) were isolated and used for flow cytometry to identify proportions of CD4⁺cells positive for either CCR4 or CD25 receptors. Both CD4⁺cell types (CD25⁺ and CCR4⁺) peaked at day 17, when clinical signs had resolved. It is proposed that the increase of circulating CD4⁺CCR4⁺ and CD4⁺CD25⁺cells most likely demonstrates the sensitization status of atopic individuals after environmental allergen challenge. Understanding of these two cell types could prompt additional research concerning therapeutic drugs for AD.

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Leptospirosis is an infectious disease caused by Leptospira interrogans sensu lato and is common in both humans and animals. In the present study, serum samples were collected from 801 dogs across all 47 prefectures in Japan, and evaluated with a microscopic agglutination test (MAT), using 5 major L. interrogans serovars (Icterohaemorrhagiae, Canicola, Autumnalis, Hebdomadis, and Australis) as antigens, and an enzyme-linked immunosorbent assay (ELISA) using recombinant OmpL1 protein as the antigen. Across all dogs tested, 217 (27.0%) and 29 (3.6%) were MAT- and ELISA-positive, respectively. However, evidence strongly suggests that MAT also detected antibodies produced by vaccination. Of 243 dogs never inoculated with any canine vaccine, 41 (16.9%) from 23 prefectures were MAT and/or ELISA positive. The most commonly detected serovar was Icterohaemorrhagiae (22 dogs, 19 prefectures). Our results suggest that there are dogs with subclinical Leptospira infection throughout Japan. To the best of our knowledge, the present study is the first nationwide survey of Leptospira infection in dogs, and the findings are relevant not only for clinical veterinary medicine but also for public health.

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Blood hyaluronic acid (HA) concentration was measured in dogs with various liver diseases to determine its relationship with histological fibrosis of the liver. The blood HA concentration significantly increased in dogs with chronic liver diseases compared with extrahepatic diseases and control. Furthermore, the median blood HA concentration in dogs with liver cirrhosis (500 μg/l; range, 151-1970 μg/l) was significantly higher than dogs with non-cirrhotic liver diseases (153 μg/l; range, 15-477 μg/l). In histochemical analysis, HA was distributed primarily in the fibrotic area in dogs with chronic liver diseases. As a conclusion, the blood HA concentration was significantly increased in dogs with chronic liver diseases, especially those with cirrhosis. Measurement of the blood HA levels of dogs with suspected liver disease can be a useful diagnostic aid for canine cirrhosis.

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A 10-year-old male Korean domestic short-haired cat was presented with refractory lower urinary tract obstruction. The cat was treated by urethral stent placement using a self-expanding nitinol intraluminal stent (Zilver^® 535 biliary stents, COOK^®, U.S.A.) subsequent with balloon expansion. Although the cat showed 2 days of transient hematuria after the stent placement, no further obstruction was occurred after the stent placement. Follow-up studies performed at monthly intervals have found no re-stenosis or particular complications, to date.

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We established a recombinant strain of Toxoplasma gondii that overexpressed programmed cell death 5 (TgPDCD5), in order to evaluate the role of endogenous TgPDCD5 in macrophage apoptosis during T. gondii infection. Immunofluorescence microscopy revealed that overproduced TgPDCD5 with a hemagglutinin tag was localized in the cytosol, which was consistent with the localization of endogenous TgPDCD5. The induced TgPDCD5-HA was recognized as an additional band by Western blot analysis, indicating successful overexpression of TgPDCD5. Secretion and release of TgPDCD5 by the parasite was also up-regulated in a time-dependent manner, which reflected its overproduction. Apoptosis due to parasite infection and interferon-gamma treatment was significantly up-regulated by the overexpression of TgPDCD5. These results suggest that endogenous TgPDCD5 plays a role in macrophage apoptosis during T. gondii infection.

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Since medium chain fatty acids (MCFAs) and their derivatives exert antimicrobial activities, their potential in controlling alimentary protozoa was examined in 12 calves infected with coccidia. Medium chain triglyceride (MCT), esterified fat of MCFAs, was administered into the rumen of 6 calves using an esophageal tube and into the abomasum of another 6 calves to ensure intestinal delivery by the function of the reticular groove reflex. Abomasal MCT inhibited the shedding of coccidial oocysts with less toxicity on ruminal protozoa. Conversely, intraruminal MCT showed negligible anticoccidial effect but toxicity on ruminal protozoa (defaunation). The anticoccidial efficacy was confirmed by abomasum feeding of MCT and not into the rumen. The results shed light on dietary fats for coccidial control.

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A case of multiple primary tumors observed in the heart base and in the lung of a 7-year-old intact female, flat-coated retriever was reported. Morphological differences between both tumors and detailed immunohistochemical study revealed that the cardiac neoplasm was as a malignant aortic body tumor and the lung tumor was a pulmonary histiocytic sarcoma. The occurrence of aortic body tumor with other primary neoplasms has been previously reported in animals suggesting that this might be a common presentation in dogs.

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Lymphoepithelial thymoma was diagnosed in a 14-year-old Samoyed dog with clinical symptoms of myasthenia gravis at 6 months of age. At necropsy, dark red-colored mass with many nodular protuberances was found in the anterior mediastinal area. Histologically, the mass consisted of solid proliferation of neoplastic cells with spindle nuclei and cytoplasm and a few lymphocytes, which is separated by an abundant fibrous and adipose tissue. Immunohistochemically, spindle cells were positive for cytokeratin, and infiltrating lymphocytes were positive for CD3. On the basis of these findings, this tumor was diagnosed as lymphoepithelial thymoma, which is morphologically similar to type A thymoma in humans.

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An adult female Aldabra giant tortoise (Geochelone gigantea) was found dead with a 2 month-history of decreased appetite. At necropsy, multiple ulcers were seen in the mucosa of stomach and colon. Histopathologically, the ulcers were characterized by extensive necrosis, hemorrhage, and marked edema, along with numerous chlamidospores and aseptate fungal hyphae. Fungal structures infiltrated into the lamina propria, submucosa and muscle layer; their invasion into blood vessels formed thrombosis. Immunohistochemically, the fungal structures were strongly positive with an antibody against Candida spp. This case is the first report on gastrointestinal candidiasis in an Aldabra giant tortoise.

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Inhibitory effects of ketoconazole (KTZ), cimetidine (CIM), and erythromycin (ERY) were examined on CYP3A activities. Midazolam 1'- and 4-hydroxylation (MDZ1'H and MDZ4H) were used to determine CYP3A activities in hepatic microsomes obtained from cats (n=4). The results showed that, all the examined drugs inhibited the reactions in a noncompetitive manner. The inhibitory constants (Ki) of KTZ were 2.80 ± 0.70 and 115 ± 28 μM for MDZ1'H and MDZ4H, respectively. Those of CIM were 3.13 and 3.27 mM and of ERY were 3.14 and 6.41 mM for MDZ1'H and MDZ4H, respectively. Mechanism-based inhibition was also examined in this study. KTZ significantly reduced MDZ reactions in a time-dependent manner; while CIM and ERY did not. Also, the effects of KTZ and CIM on the pharmacokinetics of quinidine (QUN) were studied. KTZ or CIM (10 mg/kg/day, for one week) was given orally to cats (n=5). QUN (2 mg/kg, i. v.) was injected 2 hr after the last dose of KTZ or CIM. The analysis of the obtained pharmacokinetic parameters showed that, KTZ significantly reduced total body clearance of QUN by 35%, while CIM did not. These results suggest that, KTZ inhibits CYP3A activities (both in vitro and in vivo), but CIM does not. In clinical practice, therefore, KTZ may result in inhibition based drug-drug interaction with CYP3A substrates in cat patients, whereas CIM and ERY are unlikely to lead to interaction involving CYP3A substrates.

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Visceral adipose tissue-derived serine protease inhibitor (vaspin) has been recently identified as an adipocytokine in a rat model of type 2 diabetes. Adipocytokines may directly influence the function of endothelial cells (ECs) and modulate inflammatory states. We therefore assessed the effects of vaspin on basal and TNF-α-stimulated human umbilical vein ECs. Vaspin (10-100 ng/ml, 24 hr) had no effects on both basal ECs morphology and TNF-α-induced (10 ng/ml, 24 hr) morphological damages. Vaspin did not inhibit the TNF-α (20 min) activation of JNK, p38 and NF-κB, but only slightly inhibited Akt. Furthermore, vaspin did not decrease the TNF-α (24 hr) induction of vascular cell adhesion molecule-1, intercellular adhesion molecule-1, endothelial selectin, and cyclooxygenase-2 protein expression as well as monocyte chemotactic protein-1, tissue factor, and plasmogen activator inhibitor-1 mRNA expression. The present results indicate that vaspin has no effects on normal ECs, and can not prevent TNF-α-induced inflammatory injury.

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A high-K⁺, Na⁺-deficient (isosmotic 154 mM K⁺ and 0 mM Na⁺; I-154 K⁺) solution induced contraction followed by gradual relaxation of the smooth muscles of the porcine trachea, while hyperosmotic addition of 65 mM KCl (H-65 K⁺) induced a large sustained contraction. The I-154 K⁺ solution also induced a sustained increase in [Ca²⁺]_i level. Decreases in muscle tension and increases in cellular water content were both prevented by the addition of sucrose or NaCl in the porcine trachea. An additional application of phloridzin inhibited recoveries of I-154 K⁺ solution by addition of NaCl in the porcine trachea. Addition of pyruvate or oxaloacetate recovered the I-154 K⁺ solution-induced relaxation in the porcine trachea. Although application of I-154 K⁺ solution did not affect PCr and ATP contents in the bovine trachea, the solution induced a gradual decrease of PCr content in the porcine trachea. Moreover, application of pyruvate or oxaloacetate recovered the I-154 K⁺ solution-induced decreases of PCr content in the porcine trachea. Phloridzin inhibited H-65 K⁺-induced contraction in porcine, but not in bovine trachea. In conclusion, the I-154 K⁺ solution-induced relaxation in the porcine trachea is probably due to swelling of cells and inhibition of glucose utilization. Moreover, the inhibition of glucose utilization in I-154 K⁺ medium in porcine trachea is different from that of bovine trachea.

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A full-length cDNA clone of an equine carbonic anhydrase (CA)-VI was obtained from the equine parotid gland. The cDNA sequence was 1338 bp long and was predicted to encode a 319 amino acid polypeptide with a putative signal peptide of 18 amino acids. The deduced amino acid sequence of mature CA-VI showed the similarity of 70% to those of other mammalians reported. Westernblot analysis using anti-horse CA-VI peptide detected the single band in parotid gland, and the band reduced its size by treatment with N-glycosidase F. Additionally, CA-VI protein expression was confirmed in submandicular gland and weakly in liver. In contrast, RT-PCR analysis revealed signals in the digestive tract including duodenum, jejunum, ileum, cecum and colon as well as the salivary glands. In addition, certain signals were detected in testis, thyroid gland and liver, but not in nerve tissue, skeletal muscle, spleen or lymph node.

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The adhesional function to blood vessel endothelial cells was evaluated in 2 cultured canine mammary gland tumor cells, CHMp and CHMm, derived from the primary and the metastatic lesion of the same patient. They exhibited negative (CHMp) and positive (CHMm) expression of sialyl Lewis X [sLe(x)], respectively. In the cell adhesion assay, the number of attached CHMm cells to human umbilical vein endothelial cells which expressed activated E-selectin was significantly increased. The process of cell-cell adhesion under flow conditions was observed only in CHMm cells. CHMm cells were revealed to have the adhesional ability to blood vessel endothelial cells and this adhesional ability was thought to be induced by the sLe(x)-E-selectin binding.

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A recent emerging technology, somatic cell nuclear transfer (SCNT), has been considered for conserving threatened or endangered species. Sapsaree is a native breed in Korea and has been designated as a Natural Monument. The aim of this study was to produce a Sapsaree by SCNT for breed conservation. Donor fibroblasts from a 9-year-old male Sapsaree were placed into the perivitelline spaces of enucleated in vivo matured oocytes and fused electrically. A total of 309 cloned embryos were transferred into the oviducts of 15 naturally synchronized recipients. Two recipients were diagnosed as pregnant, and each delivered one cloned puppy, both of which weighed 530 g. Overall, this study demonstrated that an endangered canine breed can be conserved by SCNT.

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