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To understand the regulatory mechanism of the secretory rhythm of GH and the involvement of melatonin (MEL) in GH regulation in cattle, daytime and nighttime profiles of GH secretion and the effect of a photic stimulation on nocturnal GH and MEL secretion were investigated in Holstein steers. Steers were kept under a constant lighting condition of 12 h of light (LIGHT; 500 lx, 0600 to 1800 h):12 h of dark (DARK; 10 lx, 1800 to 0600 h). In Exp. 1, blood was taken for 4 h at 15-min intervals during LIGHT (1100 to 1500 h) and DARK (2300 to 0300 h), respectively. The sampling was also performed from 0500 to 0900 h, with the usual light transition (light onset at 0600 h; morning sampling). In Exp. 2, steers were exposed to light (500 lx) for 1 h from 0000 to 0100 h. Plasma GH and MEL concentrations were determined by RIA and enzyme immunoassay, respectively. Both GH (P < 0.05) and MEL (P < 0.01) concentrations in plasma for 4 h during DARK were greater than those during LIGHT. On the other hand, although MEL concentrations were decreased after the light onset at 0600 during the morning, GH release was not altered. Increased GH secretion during DARK was suppressed (P < 0.01) by the 1 h of light exposure, as were MEL concentrations (P < 0.05). Pineal MEL, which was affected by the photic condition, may play an important role in the secretory rhythm of GH secretion in cattle. Key words: cattle, growth hormone, melatonin, photic stimulation, rhythm

The effects of dietary sucrose on the metabolic rate of plasma glucose and ruminal propionate as well as the change in nitrogen kinetics were examined in four mature wethers fitted with rumen fistulas in Tsukuba, Japan in 1990. Wethers were fed at 12 equal intervals daily on crushed lucerne hay cubes (1233 g DM/day), with or without 204 g/day of sucrose. Plasma urea and glucose kinetics were determined following a single intravenous injection of [I5N]urea and [U-13C]glucose respectively; and the kinetics of ruminal ammonia and propionate were determined following a single intraruminal injection of [15N]ammonium chloride and [2–13c]sodium propionate respectively. Following supplementation of sucrose to the diet, nitrogen retention was increased (P < 0·05) with a decrease in plasma urea concentration (P < 0·05) and urinary urea excretion (P < 0·05). Sucrose supplementation decreased (P < 005) the concentration and irreversible loss rate of ruminal ammonia. Urinary allantoin excretion did not change with sucrose treatment, but the flow rate of non-ammonia-nitrogen from the rumen was increased P < 0·05). The transfer rate of ruminal ammonia to plasma urea was also decreased (P < 0·01), whilst the transfer rate of plasma urea to ruminal ammonia was increased (P < 0·05) by dietary sucrose. Sucrose supplementation resulted in a higher concentration of propionate and butyrate (P < 0·05) in the rumen with no significant change in acetate or pH. The concentration of plasma glucose did not change with sucrose treatment, but the concentration of insulin, pool size (P < 0·05) and the irreversible loss rate of glucose (P < 0·01) were increased, reflecting the increase in the production rate of ruminal propionate (P < 0·05). It was concluded that the supplementation of sucrose affected the metabolism of urea and glucose in plasma via a change in ruminal production rate of ammonia and propionate, respectively. [ABSTRACT FROM PUBLISHER]

The effects of sucrose supplementation on nitrogen kinetics and energy metabolism were examined in sheep fed lucerne hay cubes using a 15N isotope dilution method and balance and respiration trials in Tsukuba, Japan in 1988. Sheep were fed lucerne hay cubes (1183 g DM/day), with or without 204 g/day sucrose, at 2 h intervals from continuous feeders. Supplementation with sucrose decreased urinary N excretion (P < 0·01), resulting in an increase in N retention from – 1·1 to + 2·0 g N/day (P < 0·01). Supplementation with sucrose resulted in no change in faeces and methane energy, a decrease in urinary energy (P < 0·01) and an increase in heat production and energy balance (P < 0·01). Sucrose supplementation also resulted in lower rumen ammonia (P < 0·05) and plasma urea concentrations (P < 0·05) and reduced urinary urea excretion (P < 0·01). The fermentation of sucrose in the rumen resulted in a decrease in rumen pH (P < 0·01) and in the acetate: propionate ratio (P < 0·05). Sucrose supplementation increased the proportion of urea transferred to the rumen (P < 0·05), non-ammonia N (NAN) concentration in the rumen (P < 0·001) and NAN flow from the rumen to the lower digestive tract (P < 0·001). Urinary allantoin excretion rate increased with sucrose supplementation (P < 0·05). The plasma glucose concentration was unchanged but plasma insulin concentration was increased with sucrose supplementation (P < 0·05). The influence of energy-rich supplements, such as sucrose, on N kinetics and the mechanism of the increase in N retention with sucrose supplementation are discussed. [ABSTRACT FROM PUBLISHER]

Abstract: The aims of the present study were to determine whether salsolinol (SAL), a dopamine-related compound, is present in the bovine posterior pituitary (PP) gland, and to clarify the effect of SAL on the secretion of prolactin (PRL) in ruminants. SAL was detected in extract of bovine PP gland using high-pressure liquid chromatography with electrochemical detection (HPLC-EC). A single intravenous (i.v.) injection of SAL (5 and 10mg/kg body weight) significantly and dose-dependently stimulated the release of PRL in goats (P <0.05). Plasma PRL levels reached a peak 10min after the injection, then gradually returned to basal values in 60–80min. The PRL-releasing pattern was similar to that in response to sulpiride (a dopamine receptor antagonist). The intracerebroventricular (i.c.v.) injection of 1mg of SAL had no significant effect on the release of PRL in calves, however, 5mg significantly stimulated the release (P <0.05) with peak values reached 30–40min after the injection. Moreover, SAL significantly stimulated the release of PRL from cultured bovine anterior pituitary cells at doses of 10−6 and 10−5 M, compared to control cells (P <0.05). Taken together, our data clearly show that SAL is present in extract of the PP gland of ruminants, and has PRL-releasing activity both in vivo and in vitro. Therefore, this endogenous compound is a strong candidate for the factor having PRL-releasing activity that has been previously detected in extract of the bovine PP gland. [Copyright &y& Elsevier]

Oxytocin (OXT) is a neuropeptide that regulates memory, emotion, stress response, and behavior in the brain. In our previous study with cattle, we demonstrated the anti-stress effect of intracerebroventricularly administered OXT on the central nervous system. However, it is important to investigate the effects of this peptide after intranasal administration, as it offers convenience and non-invasiveness for practical use. Therefore, this study investigated the effects of intranasal OXT on the behavior and autonomic nervous system of Holstein steers. The experiment followed a within-subjects design, including a total of six steers. Each steer received intranasal administration of either 1 mL of saline (SAL), 100 µg OXT (OXT100), or 200 µg OXT (OXT200). However, due to some issues, the sample size for the OXT200 group was reduced to five. After these treatments, we conducted electrocardiography recordings to analyze heart rate variability (HRV) and also made behavioral observations for 90 min. OXT200 tended to increase the time spent ruminating while lying down (Steel's multiple comparison test; P=0.053). In contrast, OXT treatment did not affect HRV indices. In conclusion, the current OXT dosage did not significantly affects behavior or the autonomic nervous system. However, the observed tendency to increase rumination may suggest a central effect of OXT.

Biopsy is a commonly used method for determining pathological diagnoses by directly using human tissues and cells. Biopsies are widely used to determine disease progression and treatment efficacy. Although organs and tissues are usually obtained by sacrifice during animal experiments, it is theoretically possible to use the same biopsy techniques in humans. In the present study, we examined the feasibility of performing four repeated liver biopsies in a spontaneous metabolic syndrome mouse model. Even though a small number of mice died accidently, most mice were able to undergo four liver biopsies without significant adverse events. We also performed three liver biopsies in mouse liver tumor carcinogen models at 4, 8, and 12 weeks of age. In addition to the sample collected at 16 weeks of age during sacrifice, we successfully collected four liver samples from the same mice at different stages of disease progression. The application of this liver biopsy technique might make it possible for direct evaluation of pathological conditions in the same individual over time, thereby reducing the number of experimental animals., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: KOICHI TSUNEYAMA reports financial support was provided by JSPS-KAKENHI. KOICHI TSUNEYAMA reports a relationship with JSPS-KAKENHI that includes: funding grants., (© 2023 The Authors.)

According to previous clinical studies, the prevalence of non-alcoholic fatty liver disease (NAFLD) is higher in men than women only during the reproductive age. Animal models of NAFLD that reflect sex differences in humans have not been established. In this study, we examined sex differences in the hepatic lesions of Tsumura Suzuki obese diabetes (TSOD) and db/db mice, which are representative genetic models of NAFLD. Male and female TSOD and db/db mice were fed with a normal diet and tap water ad libitum. Six male and female mice of each strain were sacrificed at the ages of 3 and 9 months, respectively, and serum biochemical, pathological, and molecular analyses were performed. Serum aspartate aminotransferase (AST) levels were significantly higher in male than female mice of both strains at the age of 3 months; however, at 9 months, significant sex differences were not observed. Similarly, alanine aminotransferase (ALT) levels were significantly higher in male mice than in female TSOD mice at the age of 3 months; however, at 9 months, significant sex differences were not observed. Image analysis of histological slides revealed that the frequency of the steatotic area was significantly higher in male than female db/db mice at the age of 3 months; however, significant sex differences were not observed at 9 months. The frequency of Sirius red-positive fibrotic area was significantly higher in male than female mice in both strains at the age of 3 months; however, significant sex differences were not observed at 9 months. Serum AST and ALT levels and hepatic steatosis and fibrosis in TSOD and db/db mice showed age-dependent sex differences consistent with those observed in human NAFLD. These mice may be suitable for studying sex differences of the disease., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 Dungubat et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Male Tsumura-Suzuki Obese Diabetes (TSOD) mice, a spontaneous metabolic syndrome model, develop non-alcoholic steatohepatitis and liver tumors by feeding on a standard mouse diet. Nearly 70% of liver tumors express glutamine synthetase (GS), a marker of hepatocellular carcinoma. In contrast, approximately 30% are GS-negative without prominent nuclear or structural atypia. In this study, we examined the characteristics of the GS-negative tumors of TSOD mice. Twenty male TSOD mice were sacrificed at 40 weeks and a total of 21 tumors were analyzed by HE staining and immunostaining of GS, liver fatty acid-binding protein (L-FABP), serum amyloid A (SAA), and beta-catenin. With immunostaining for GS, six (29%) tumors were negative. Based on the histological and immunohistological characteristics, six GS-negative tumors were classified into several subtypes of human hepatocellular adenoma (HCA). One large tumor showed generally similar findings to inflammatory HCA, but contained small atypical foci with GS staining and partial nuclear beta-catenin expression suggesting malignant transformation. GS-negative tumors of TSOD mice contained features similar to various subtypes of HCA. Different HCA subtypes occurring in the same liver have been reported in humans; however, the diversity of patient backgrounds limits the ability to conduct a detailed, multifaceted analysis. TSOD mice may share similar mechanisms of HCA development as in humans. It is timely to review the pathogenesis of HCA from both genetic and environmental perspectives, and it is expected that TSOD mice will make further contributions in this regard.

Non-alcoholic steatohepatitis (NASH) results from non-alcoholic fatty liver disease (NAFLD) via multiple-parallel events, including hepatic triglyceride accumulation, oxidative stress, and inflammation. The complex interaction between the liver and multiple other organs is involved in NASH development. Although spleen-derived humoral factors can directly contribute to NAFLD/NASH onset via the portal vein, the status of the spleen in the early stage of NASH remains unknown. Here, our aim was to investigate whether splenocytes may exacerbate NASH via the generations of reactive oxygen species (ROS) and proinflammatory cytokines. Iron accumulation was observed in the spleen but not the liver, and the proportion of phagocytic macrophages increased in the spleen of Tsumura Suzuki Obese Diabetes (TSOD) mice showing histological characteristics of NASH in the early stage. The splenocytes generated moderate amounts of ROS and released high amounts of tumor necrosis factor (TNF)-α in response to lipopolysaccharide, indicating excessive inflammatory cytokine released by activated macrophages in iron-accumulating spleens. Our results suggest that iron-accumulating splenocytes can easily induce inflammation and contribute to exacerbate NASH via the portal vein. Thus, the regulation of iron metabolism in the spleen should be considered in the development of novel therapeutic targets against NASH.

Although diabetes mellitus (DM) is a well-known risk factor for hepatocellular carcinoma (HCC), the underlying mechanisms have not yet to be defined. We previously reported that DIAR mice fed with standard murine diet developed type 1 diabetes and HCC at age of 16 weeks old with a neonatal streptozotocin treatment (n-STZ). Because DIAR mice did not manifest obesity nor develop steatohepatitis, hyperglycemia with streptozotocin trigger or streptozotocin alone might turn on the hepato-carcinogenesis. An insulin-recruitment to DIAR-nSTZ mice showed an increased frequency of HCC during the first 12 weeks of age, although the diabetic indications notably improved. To elucidate the role of hyperglycemia in hepato-carcinogenesis, we performed a head-to-head comparative study by using 4CS mice and DIAR mice with n-STZ treatment. Newborn 4CS mice and DIAR mice were divided into STZ treated group and control group. The blood glucose levels of DIAR-nSTZ mice increased at age of eight weeks, while that of 4CS-nSTZ mice were maintained in the normal range. At eight weeks old, three out of five DIAR-nSTZ mice (60%) and one out of ten 4CS-nSTZ mice (10%) developed multiple liver tumors. At age of 12 weeks old, all eight of DIAR-nSTZ mice (100%) and two of 10 4CS-nSTZ mice (20%) developed multiple liver tumors. At 16 weeks old, all animals of DIAR-nSTZ and 4CS-nSTZ mice occurred liver tumors. DIAR-nSTZ showed hyperglycemia and HCC, and 4CS-nSTZ developed HCC without hyperglycemia. These results were interpreted that the onset of HCC maybe not related to the presence or absence of hyperglycemia but nSTZ treatment. On the other hand, since the carcinogenesis of 4CS-nSTZ is delayed compared to DIAR-nSTZ, hyperglycemia may play a role in the progression of carcinogenesis. Histologically, the liver tumor appeared irregularly trabecular arrangements of hepatocytes with various degrees of nuclear atypia. By immunohistochemical analyses, all liver tumors showed positive staining of glutamine synthetase (GS), an established human HCC marker. The expression pattern of GS was divided into a strong diffuse pattern and weak patchy pattern, respectively. The liver tumor showing the weak GS-patchy pattern expressed biliary/stem markers, EpCAM, and SALL4, partially. Because 4CS-nSTZ mice did not show any metabolic complications such as gaining body weight or high blood glucose level, it is a unique animal model with a simple condition to investigate hepatic carcinogenesis by excluding other factors., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

This study was conducted to investigate the effect of tryptophan (TRP) supply on the thermoregulatory responses via brain serotonin (5-HT) in cattle. In period 1, 12 Holstein steers were kept under a constant room temperature (22°C) and were administered the intravenous (i.v.) infusion of saline or TRP (38.5 mg/kg/2 h). Changes in rectal temperature (RT), 5-HT concentration in the cerebrospinal fluid (CSF), and other factors involved in thermoregulation were measured. In period 2, the steers received the same treatments as in period 1; however, the room temperature was elevated from 22°C to 33°C during i.v. infusion and maintained at 33°C for 3 h. 5-HT concentration in CSF increased following TRP infusion in both periods, and RT significantly decreased following TRP infusion only in period 2. The effect of TRP on respiration rate and plasma prolactin and total triiodothyronine concentrations was not significant. These results suggest that increase in TRP supply can attenuate increase in RT in response to acute heat stress through the increase in brain 5-HT, followed by presumable increase in evaporative heat loss from the skin surface in cattle. It is possible that the increase in peripheral blood TRP metabolites could also participate in the hypothermic effect of TRP., (© 2018 Japanese Society of Animal Science.)

The effects of l-DOPA, a precursor of dopamine (DA), and sulpiride, a D 2 -type DA receptor blocker, on growth hormone (GH) and prolactin (PRL) secretion were investigated in steers. Eight Holstein steers (212.8 ± 7.8 kg body weight) were used. Lighting conditions were 12:12 L:D (lights on: 06.00-18.00 hours). Blood samplings were performed during the daytime (11.00-15.00 hours) and nighttime (23.00-03.00 hours). Intravenous injections of drugs or saline were performed at 12.00 hour for the daytime and 00.00 hour for the nighttime, respectively. Plasma GH and PRL concentrations were determined by radioimmunoassay. l-DOPA did not alter the GH secretion when it was injected at 12.00 hour (spontaneous GH level at its peak). On the other hand, l-DOPA increased GH secretion at 00.00 hour (GH level at its trough). Injection of sulpiride suppressed GH secretion at 12.00 hour but did not affect GH levels at 00.00 hour. l-DOPA inhibited and sulpiride stimulated PRL release during both periods. These results suggest that dopaminergic neurons have stimulatory action on GH secretion and inhibitory action on PRL secretion in cattle. In addition, injection time should be considered to evaluate the exact effects on GH secretion due to its ultradian rhythm of GH secretion in cattle., (© 2017 Japanese Society of Animal Science.)

We previously reported that Tsumura-Suzuki obese diabetic (TSOD) mice, a polygenic model of spontaneous type 2 diabetes, is a valuable model of hepatic carcinogenesis via non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). One of the characteristics of tumors in these mice is the diffuse expression of glutamine synthetase (GS), which is a diagnostic marker for hepatocellular carcinoma (HCC). In this study, we performed detailed histopathological examinations and found that GS expression was diffusely positive in >70% of the hepatic tumors from 15-month-old male TSOD mice. Translocation of β-catenin into nuclei with enhanced membranous expression also occurred in GS-positive tumors. Small lesions (<1 mm) in GS-positive cases exhibited dysplastic nodules, with severe nuclear atypia, whereas large lesions (>3 mm) bore the characteristics of human HCC, exhibiting nuclear and structural atypia with invasive growth. By contrast, the majority of GS-negative tumors were hepatocellular adenomas with advanced fatty change and low nuclear grade. In GS-negative tumors, loss of liver fatty acid-binding protein expression was observed. These results suggest that the histological characteristics of GS-positive hepatic tumors in TSOD mice resemble human HCC; thus, this model may be a useful tool in translational research targeting the NAFLD/NASH-HCC sequence.

This study was conducted to investigate the possibility of suppression of stress-induced cortisol (CORT) secretion by tryptophan (TRP) administration and to better understand its regulatory mechanisms by using a noradrenaline (NA) injection into the third ventricle (3V) as a stress model in cattle. A total of 25 Holstein steers with a cannula in the 3V were used. First, the increase in CORT secretion was observed following a NA injection into the 3V in a dose-dependent manner, verifying the appropriateness of this treatment as a stress model of CORT secretion (Experiment 1). The effect of prior-administration of TRP into peripheral blood with a dose that has been demonstrated to increase brain 5-hydroxytryptamine levels on the elevation of plasma CORT induced by NA or corticotropin-releasing hormone (CRH) was then examined (Experiment 2). The prior administration of TRP suppressed NA-induced, but not CRH-induced, CORT elevation. These results suggest that an increase in TRP absorption into peripheral blood could suppress the stress-induced CORT secretion in cattle via the attenuation of the stimulatory effect of NA on the hypothalamic CRH release., (© 2015 Japanese Society of Animal Science.)

Outer-arm dynein is the main engine providing the motive force in cilia. Using three-dimensional tracking microscopy, we found that contrary to previous reports Tetrahymena ciliary three-headed outer-arm dynein (αβγ) as well as proteolytically generated two-headed (βγ) and one-headed (α) subparticles showed clockwise rotation of each sliding microtubule around its longitudinal axis in microtubule corkscrewing assays. By measuring the rotational pitch as a function of ATP concentration, we also found that the microtubule corkscrewing pitch is independent of ATP concentration, except at low ATP concentrations where the pitch generated by both three-headed αβγ and one-headed α exhibited significantly longer pitch. In contrast, the pitch driven by two-headed βγ did not display this sensitivity. In the assays on lawns containing mixtures of α and βγ at various ratios, the corkscrewing pitch increased dramatically in a nonlinear fashion as the ratio of α in the mixture increased. Even small proportions of α-subparticle could significantly increase the corkscrewing pitch of the mixture. Our data show that torque generation does not require the three-headed outer-arm dynein (αβγ) but is an intrinsic property of the subparticles of axonemal dyneins and also suggest that each subparticle may have distinct mechanical properties., (Copyright © 2015 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

Background and Aim: Non-alcoholic steatohepatitis (NASH) is the hepatic manifestation of metabolic syndrome (MS). Monosodium glutamate (MSG)-treated ICR mice is a useful model of MS and NASH, but it shows the different patterns of steatosis from human NASH. Because inbred aged DIAR (ddY, Institute for Animal Reproduction) mice spontaneously show the similar pattern of steatosis as NASH, we analyzed their liver pathology after administering MSG., Methods: MSG-treated DIAR mice (DIAR-MSG) and untreated DIAR mice (DIAR-controls) were sacrificed and assessed histopathologically at 29, 32, 40, 48, and 54 weeks of age. The NASH activity score, body mass index, blood glucose level, and oral glucose tolerance test were also assessed., Results: The body mass index and blood glucose levels of DIAR-MSG were significantly higher than controls. The oral glucose tolerance test revealed a type 2 diabetes pattern in DIAR-MSG. The livers of DIAR-MSG mice showed macrovesicular steatosis, lobular inflammation with neutrophils, and ballooning degeneration after 29 weeks. At 54 weeks, mild fibrosis was observed in 5/6 DIAR-MSG and 2/5 DIAR-control mice. In imaging mass spectrometry analysis, cholesterol as well as triglyceride accumulated in the liver of DIAR-MSG mice. Atypical liver nodules were also observed after 32 weeks in DIAR-MSG, some with cellular and structural atypia mimicking human hepatocellular carcinoma. The NASH activity score of DIAR-MSG after 29 weeks was higher than that of control mice, suggesting the development of NASH., Conclusions: DIAR-MSG had NASH-like liver pathology and liver nodules typically associated with MS symptoms. DIAR-MSG provides a valuable animal model to analyze NASH pathogenesis and carcinogenesis., (© 2014 Journal of Gastroenterology and Hepatology Foundation and Wiley Publishing Asia Pty Ltd.)

Objective: The operative strategies for retrograde acute type A aortic dissection with a primary intimal tear remain controversial because resection of an intimal tear via a median sternotomy is difficult. We evaluated the frozen elephant trunk technique for treating this type of aortic dissection., Methods: The frozen elephant trunk technique was used for acute retrograde type A aortic dissection with a primary intimal tear in the descending aorta in 25 consecutive patients (16 men, 9 women; median age, 64 years) seen between 1997 and 2011 at the Hiroshima City Asa Hospital. Three patients had Marfan syndrome. Fourteen patients had ischemia of the lower half of the body, 2 had stroke, 3 had coronary ischemia, and 10 had pericardial tamponade., Results: There were no hospital deaths. Postoperative complications occurred in 2 patients, including stroke and mediastinitis. Two late deaths occurred over a median follow-up period of 58 months (12-169 months), one caused by stroke and the other by colon cancer. Two patients needed additional operations: one required a second operation for descending aorta replacement because of ulceration caused by the stent graft, and the other required an endovascular stent graft because of dilatation of the descending aorta. Computed tomography follow-up scans revealed that the false lumen was completely thrombosed and obliterated in all 25 patients with stent grafts., Conclusions: The frozen elephant trunk technique for retrograde type A aortic dissection yielded acceptable outcomes, despite the high preoperative morbidity, and might improve the prognosis of dissection., (Copyright © 2014 The American Association for Thoracic Surgery. Published by Mosby, Inc. All rights reserved.)

To clarify endocrine responses to psychological stressors in cattle, the effects of isolation from familiar peers on plasma prolactin (PRL) and cortisol (CORT) concentrations, and the effect of 3,4-dihydroxy-L-phenylalanine (L-DOPA), a precursor of dopamine (DA), on stress-induced PRL secretion were determined in Holstein steers. First, the potency of peripheral L-DOPA administration on attenuation of central DA levels was confirmed. Cerebrospinal fluid (CSF) collected from a chronic cannula in the third ventricle and plasma were sampled 1 h before and 3 h after intravenous injection of L-DOPA (100 mg/head). DA concentrations in CSF increased just after L-DOPA injection with subsequent decrease in PRL secretion. Injection of L-DOPA increased CORT secretion. Second, one experimental steer was isolated in its stall by removing its peers for 2 h with or without- pre-injection of L-DOPA. The concentration of PRL was elevated by isolation treatment, whereas the effect of isolation on CORT concentration could not be detected. The increase in PRL concentration after isolation was abolished by pre-injection of L-DOPA. These results suggest that PRL responds to isolation and that DA neurons in the central nervous system may regulate stress-induced PRL secretion in steers., (© 2013 Japanese Society of Animal Science.)

This study was conducted to investigate physiological and behavioral effects of the exposure to plant-derived odorants in Holstein steers (8 month of age) under the acute stress of social isolation in a novel environment. Each steer was tethered in a new room alone and exposed to one of three odorants: a mixture of equal amounts of trans-2-hexenal and cis-3-hexenol (green odor, GO; n = 5), essential oil of grapefruits (EOG; n = 5) and solvent (SOL; n = 6). Behaviors were recorded and blood samples were taken at intervals of 10-15 min for 6 h. Compared with SOL, exposure to GO or EOG decreased the frequency of vocalization (P < 0.05), shortened the latency period before the onset of rumination (P < 0.05) and increased the duration of rumination (P < 0.05). Plasma cortisol concentrations in steers exposed to plant-derived odorants (GO and EOG) during 90-130 min from commencement of sampling were significantly lower (P < 0.05) compared with SOL. These results suggest that appeasing effects of plant-derived odorants on behavioral and physiological stress responses previously reported in laboratory rodents could be also be expected in steers., (© 2012 The Authors. Animal Science Journal © 2012 Japanese Society of Animal Science.)

To shed light on the role of central oxytocin (OXT) in regulating social behavior in cattle, the impact of intracerebroventricularly administered OXT agonist, carbetocin (CBT), on the social behavior of a group of familiar steers was investigated. In the first experiment, we determined the dose response of intracerebroventricularly administered CBT (0.5, 5 or 50 nmol) on plasma cortisol level and behavior using 7 steers aged from 6 to 10 months. Five of the steers were assigned to the second experiment. CBT (50 or 200 nmol/200 μl) in artificial cerebrospinal fluid (aCSF) or aCSF (200 μl) was injected into the third ventricle. Immediately after the injection, the animal and two peers were taken outside to the adjacent paddock. Thirty minutes later, maintenance and social behaviors of the animal were observed for 2 hr. CBT had no effect either on the basal cortisol level or on the maintenance and the abnormal behavior in steers with their movement restricted by a stanchion stall in the first experiment. However, in the same steers with no movement restrictions in the second experiment, CBT facilitated lying, probably because of its sedative effect via OXT receptor activation, which disturbed some aspects of social behavior. These results suggest that central OXT receptor activation might not affect social behavior itself among "familiar members", because the stimulation of the central OXT system by intracerebroventricular administration of CBT did not facilitate social behavior between familiar steers.

We reported simple partial root remodeling using a graft trimmed twin "U" shape for extensive dissection into the right/non-coronary sinuses with acute type A aortic dissection with cardiac tamponade, acute myocardial infarction on the right coronary artery and aortic regurgitation. After total arch replacement with frozen elephant trunk technique for distal dissection, partial root remodeling on the non and right coronary sinuses was performed. Postoperative computed tomography (CT) showed well shaped Valsalva sinuses and aortic regurgitation completely disappeared. This technique might become an alternative procedure for aortic dissection severely involving Valsalva sinuses without dilatation on the aortic root.

Core 2 β-1,6-N-acetylglucosaminyltransferase (C2GnT) forms an N-acetylglucosamine branch in O-glycans (core 2 O-glycans) of cell surface glycoproteins. C2GnT-expressing bladder tumors acquire highly metastatic phenotypes by surviving longer in host blood circulation. However, the detailed mechanisms underlying this increased survival remain unclear. In this study, we report that the expression of C2GnT in bladder tumors positively correlates with tumor progression and that bladder tumor cell-surface mucin 1 (MUC1) carrying core 2 O-glycans plays an important role in the evasion from natural killer (NK) cell attack. In C2GnT-expressing bladder tumor cells, heavily core 2 O-glycosylated MUC1 carries poly-N-acetyllactosamine in its O-glycans and galectin-3 binds to MUC1 through this poly-N-acetyllactosamine. The binding of galectin-3 to poly-N-acetyllactosamine in MUC1 core 2 O-glycans attenuates the interaction of the tumor cells with NK cells and interferes with the access of tumor necrosis factor-related apoptosis-inducing ligand to the tumor cell surface. These effects of MUC1 carrying core 2 O-glycans on NK cell attack facilitate C2GnT-expressing tumor cells to evade NK cell immunity and survive longer in host blood circulation. We reveal that MUC1 carrying core 2 O-glycans thus functions as a molecular shield against NK cell attack, thereby promoting bladder tumor metastasis.

Growth hormone (GH) secretion regularity and the effects of lighting condition and GH-releasing hormone (GHRH) on GH release were determined in steers. First, steers were kept under 12:12 L : D conditions (light: 06.00-18.00 hours). The animals were then subjected to a 1-h advancement in lighting on/off conditions (05.00 and 17.00 hours, respectively). Blood was sampled for 24 h at 1-h interval on the seventh day of each condition. Second, GHRH was injected intravenously (IV) at 12.00 and 00.00 hours under 12:12 L : D and blood was sampled at 15-min interval for 4-h (1 h before and 3 h after the injection). Plasma GH concentrations were measured by a radioimmunoassay. Periodicity of GH secretory profile was calculated by power spectrum analysis using the maximum entropy method. Plasma GH concentrations showed a characteristic pattern consisting of four distinct peaks. Mean periodicity of GH secretory profile was 5.7 h, and it was not altered by any change in lighting conditions. IV injection of GHRH increased GH secretion during the day and night. The increase in GH secretory volume after GHRH injection during the night was equal to that during the day. The present results suggest that GH secreted from the anterior pituitary have regularity in steers., (© 2011 The Authors. Animal Science Journal © 2011 Japanese Society of Animal Science.)

Purpose: We have introduced a new surgical approach for extended thoracic aortic repair, anterolateral thoracotomy with partial sternotomy (ALPS)., Description: The surgical approach to the chest was made via left anterolateral thoracotomy and lower median sternotomy through the third or fourth intercostal space. All cannulations (arterial, venous, venting, and cardioplegia) could be easily performed using a retractor in this approach., Evaluation: From November 2005 to December 2010, we performed surgical treatment in 12 patients by employing the ALPS approach for a complex, extended thoracic aortic diseases with different pathologies, i.e., arteriosclerotic aneurysms in 5, acute type B dissection in 5, and chronic type B dissection in 2 patients. One patient died in the hospital, and 1 had temporary spinal cord injury., Conclusion: The ALPS approach might become an alternative for a complex, extended thoracic aortic replacement.

As a preliminary step to elucidate the involvement of central neurotransmitters in the dip in voluntary feed intake during the perinatal period in cows, we investigated the effect of intracerebroventricular (ICV) administration of oxytocin, prolactin-releasing peptide (PrRP), or corticotropin-releasing hormone (CRH), the central functions of all of which undergo drastic changes during the perinatal period, on feed intake in steers. Thirty minutes before the onset of feeding, the treatment solution was injected into the third ventricle through an implanted cannula, and feeding-related behaviors were observed for 1 h after the onset of feeding. Neither ICV oxytocin (5 and 50 μg) nor PrRP (2 and 20 nmol) reduced feed intake (n=6). Twenty nanomoles of bovine CRH noticeably inhibited feeding behavior compared with vehicle treatment (n=5, p<0.05). Fifty micrograms of oxytocin reduced latency to the first water access after feeding onset (p<0.1), which may be because of the stimulation of arginine vasopressin V1b receptor by the high dose of oxytocin. We conclude that CRH inhibits feeding behavior by its central action in this species, although this could also be an indirect effect due to the increased expression of abnormal behaviors caused by CRH. Central administration of neither oxytocin nor PrRP reduced feed intake in steers. Although the effects of sex steroids need to be examined, it appears that increased activity of oxytocin, and possibly PrRP, during the perinatal period does not contribute to the dip in voluntary feed intake in this species. On the other hand, it makes sense that suppressed central CRH activity during the perinatal period should act in the direction of maintaining or even increasing food intake to aid a steady supply of energy to the fetus or offspring. We thus speculate that CRH is not a prime candidate involved in the dip in voluntary feed intake during the perinatal period in cows., (Copyright © 2011 Elsevier Inc. All rights reserved.)

We reported successful distal repair using the frozen elephant trunk technique to treat a mycotic aneurysm extending from the ascending aorta to the descending aorta. A blood culture sample was positive for Escherichia coli, and total arch replacement with a rifampicin-bonded graft covered the omental pedicle flap.

Objective: The aimed to describe the frozen elephant trunk (FET) technique and partial remodeling (PR) for acute type A aortic dissection (ATAAD), considering the long-term prognosis on the basis of our 13 years of experience., Methods: There were 80 consecutive patients (mean age: 66.4 years) with an FET and PR technique for ATAAD between September 1997 and February 2010. We indicated a PR for all 80 patients without dilatation of the sinuses and a FET for 20 patients with a distal entry in the descending aorta, 14 patients with a dilatation more than 4cm on the distal arch and 46 patients with a narrow true lumen younger than 70 years with a narrow true lumen. During moderate hypothermic circulation with selective cerebral perfusion and distal perfusion from the femoral artery, a stent graft (mean diameter: 27.7mm, mean length: 9.9mm, mean distal depth: thoracic vertebra (Th) 6.0th) was inserted through the transected proximal aortic arch. The plication of the sinotubular junction (N=42) or partial remodeling for right and/or non-coronary cusp (N=38) was performed after total arch replacement with a four-branched prosthesis., Results: Four patients died in hospital. Early morbidity included two (2.5%) strokes but no spinal cord injury. In long-term follow-up (mean 94.6 months), five patients died of non-aortic events and two re-operations (Bentall and stent grafting to the descending aorta) were required. No patients had patent false lumen on the stent graft and residual aortic regurgitation, according to late follow-up computed tomography (CT) and echogram. The 10-year survival was 75% and the overall 10-year re-operation free rate on the thoracic aorta was 95%., Conclusion: FET and modified PR techniques could be effective for improving the long-term outcome on the distal and proximal aorta in an ATAAD., (Crown Copyright © 2011. Published by Elsevier B.V. All rights reserved.)

The O-glycan branching enzyme, core2 β-1,6-N-acetylglucosaminyltransferase (C2GnT), forms O-glycans containing an N-acetylglucosamine branch connected to N-acetylgalactosamine (core2 O-glycans) on cell-surface glycoproteins. Here, we report that upregulation of C2GnT is closely correlated with progression of bladder tumours and that C2GnT-expressing bladder tumours use a novel strategy to increase their metastatic potential. Our results showed that C2GnT-expressing bladder tumour cells are highly metastatic due to their high ability to evade NK cell immunity and revealed the molecular mechanism of the immune evasion by C2GnT expression. Engagement of an NK-activating receptor, NKG2D, by its tumour-associated ligand, Major histocompatibility complex class I-related chain A (MICA), is critical to tumour rejection by NK cells. In C2GnT-expressing bladder tumour cells, poly-N-acetyllactosamine was present on core2 O-glycans on MICA, and galectin-3 bound the NKG2D-binding site of MICA through this poly-N-acetyllactosamine. Galectin-3 reduced the affinity of MICA for NKG2D, thereby severely impairing NK cell activation and silencing the NK cells. This new mode of NK cell silencing promotes immune evasion of C2GnT-expressing bladder tumour cells, resulting in tumour metastasis.

Purpose: Invadopodia (protrusions of the plasma membrane formed by invasive tumor cells) have an essential role in bladder tumor invasion. To understand the process of bladder tumor invasion it is crucial to investigate the molecular mechanisms of invadopodia formation. We found that invasive bladder tumor cells express FBP17. In this study we examined the role of FBP17 in bladder tumor cell invadopodia formation and invasion., Materials and Methods: We used the 3 bladder tumor cell lines YTS-1, T24 and RT4 (ATCC®), and primary culture of bladder tumors from patients. Cells were stained with phalloidin for invadopodia formation. FBP17 knockdown cells were tested for invadopodia formation and subjected to invasion assay using a Transwell® cell culture chamber. We also examined the role of the extended FER-CIP4 homology and Src homology 3 domains of FBP17 in invadopodia formation in FBP17 mutant constructs., Results: Invadopodia formation was observed in invasive bladder tumor cells and FBP17 was localized to invadopodia in invasive cells. FBP17 knockdown decreased invadopodia formation in invasive cells to 13% to 14% (p <0.0005) and decreased their invasive capacity to 14% to 16% (p <0.001). The extended FER-CIP4 homology and Src homology 3 domains of FBP17 were necessary for invadopodia formation and invasion., Conclusions: Invadopodia formation requires membrane deformation activity and recruitment of dynamin-2 mediated by FBP17. FBP17 has a critical role in the process of bladder tumor cell invasion by mediating invadopodia formation., (Copyright © 2011 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.)

Purpose: We describe the long-term efficacy of early entry closure for acute type B aortic dissection by open stent grafting based on long-term results and changes in the false lumen on enhanced computed tomography (CT)., Methods: We performed open stent grafting for acute type B aortic dissection in 28 28 patients between 1998 a 2008. They included 14 patients with complicated type B aortic dissection (11 patients with limb and/or visceral ischemia and 4 patients with impending rupture). Uncomplicated type B aortic dissection was present in 14 patients, including 4 patients with an ulcer-like projection and 10 patients who had stable dissection with a patent false lumen., Results: Two patients who had the ischemic type died within 30 days. Postoperative complications occurred in two patients, including one case of paraplegia and one of mediastinitis. Two more deaths occurred during a median follow-up period of 45 months (range 6-114 months), with one related to open stent-graft infection and one due to pneumonia. The overall survival rate 5 years after open stent grafting was 77%. According to follow-up by CT, the false lumen was completely thrombosed and obliterated in all 26 patients with stent grafts., Conclusion: Early entry closure for both complicated and uncomplicated acute type B aortic dissection with an open stent graft has an acceptable early outcome and provides a high aortic remodeling rate.

Some evidence suggests that there might be a species difference in the effect of intracerebroventricularly administered (ICV) prolactin-releasing peptide (PrRP) between rodents and sheep. We compared the levels of cortisol (CORT) and prolactin (PRL), rectal temperature (RT) and behavioral responses to ICV bovine PrRP (bPrRP) in steers. ICV bPrRP (0.2, 2 and 20 nmol/200 µL) tended to evoke a dose-related increase in CORT concentrations and 0.2 nmol of bPrRP induced transient increase in PRL concentrations. A significant time-treatment interaction was observed for the percent change of CORT (P<0.05) and PRL (P<0.05) from pre-injection value. The time-treatment interaction for changes in RT was not significant (P=0.50). There tended to be a difference among the four treatments in terms of maximum change in RT from the pre-injection value between 0 and 90 min (P<0.1). Stress-related behavioral signs were not observed in the present experiment. These findings indicate that ICV bPrRP increased CORT and PRL levels, suggesting that central PrRP might participate in controlling the hypothalamo-pituitary-adrenal axis and PRL release in cattle, unlike sheep. In contrast, central PrRP is unlikely to be involved in controlling the behavior of this species because ICV bPrRP did not induce marked changes in their behavior., (© 2011 The Authors. Journal compilation © 2011 Japanese Society of Animal Science.)

Objectives: This study describes the long-term safety and effectiveness of extended aortic arch replacement with the frozen elephant trunk technique from our 12 years of experience., Methods: Between September 1997 and September 2008, 156 patients (mean age 67.9 years) with different pathologies from the aortic arch to the extended descending aorta in 100 dissections (acute A/acute B/chronic B=66/26/8) and 56 thoracic arteriosclerotic aneurysms (TAAs) had the frozen elephant technique performed upon them. During moderate hypothermic circulation with selective cerebral perfusion, the stent graft was inserted through the transected proximal aortic arch with trans-oesophageal echo guidance., Results: Six patients (3.2%) (acute A/acute B/chronic B/TAA=3/2/0/1) died in hospital. Postoperative morbidity induced four (2.6%) strokes (acute A/acute B/chronic B/TAA=2/0/0/2) and three (2.0%) spinal injuries (paraplegia in two and transient paraparesis in one) (acute A/acute B/chronic B/TAA=0/0/1/2). In the long-term follow-up (mean 63.3 + or - 39.2 months, maximum 144 months) 16 patients died. The survival rate was 99.3%, 86.5% and 74.9% at 1, 5 and 10 years, respectively. An additional operation was performed in 15 (9.4%) (ascending aorta/aortic root/descending aorta/abdominal aorta=1/2/5/7) including three stent-graft-related events (2.1%), and the additional repair proved successful. A follow-up computed tomography (CT) image was available for 96.0% (143/149) of patients who survived longer than 12 months. The size of false lumen or aneurysm increased in four patients, was unchanged in 20 patients (14.0%), shrank in 66 (46.2%) and was completely obliterated in 55 (37.1%)., Conclusions: The frozen elephant technique could be an attractive treatment for extended aortic arch disease to the extended descending aorta for acute aortic dissection as well as arteriosclerotic aneurysm., (Crown Copyright 2010. Published by Elsevier B.V. All rights reserved.)

To clarify the role of serotonin (5-HT) in the regulatory mechanism of L-tryptophan (TRP)--induced growth hormone (GH) secretion in cattle, changes in 5-HT concentrations in the cerebrospinal fluid (CSF) in the third ventricle (3V) and GH in plasma before and after the peripheral infusion of TRP were determined simultaneously. The direct effect of TRP on GH release from the dispersed anterior pituitary cells was also assessed. A chronic cannula was placed in 3V by stereotaxic surgery, then CSF and blood were withdrawn under physiological conditions. TRP (38.5 mg/kg BW) was infused through an intravenous catheter from 12.00 to 14.00 hours and CSF and blood sampling were performed from 11.00 to 18.00 hours at 1-h intervals. The concentration of 5-HT in CSF was determined by high-performance liquid chromatography with electrochemical detection. GH, melatonin (MEL), and cortisol (CORT) concentrations were measured by radio-immunoassay and enzyme-immunoassay. Concentrations of 5-HT were increased by TRP infusion. The TRP infusion significantly increased GH release. On the other hand, TRP did not stimulate GH release from the bovine pituitary cells. MEL and CORT concentrations were not altered by TRP infusion. These results suggest that TRP induced GH release via the activation of serotonergic neurons in cattle.

Objectives: The objective of this report is to elucidate the feasibility of the frozen elephant trunk technique as a one-stage operation for extensive arteriosclerotic aneurysms and to investigate the long-term durability and efficacy of this procedure from our 11 years of experience., Methods: The subjects were 58 consecutive patients who electively received the frozen elephant trunk technique for arteriosclerotic aneurysms involving the aortic arch and the descending aorta between September 1997 and September 2008. Concomitant procedures included 15 coronary artery bypass grafts, 2 aortic valve replacements, 1 aortic root replacement, and 3 maze procedures. The stent graft was delivered to the seventh thoracic vertebra level (Th7) in 22, Th8 in 26, Th9 in 8, and Th10 in 2 patients. Cerebrospinal fluid drainage was administered preoperatively in 10 (17.2%) patients., Results: Operative mortality within 30 days was 0 of 58. There was 1 in-hospital death. Perioperative morbidity included strokes and spinal cord injury in 2 (3.4%) patients each. During the mean follow-up period of 54.2 +/- 36.9 months, there were 9 (15.5%) late deaths, and 7 (12.1%) patients required additional intervention. The 8-year survival was 65.5%, the overall 8-year aortic event free survival was 72.8%, and the 8-year event free survival on the site of the stent graft was 94.8%. A follow-up computed tomographic image was available for 86.2% (50/58) of patients who survived longer than 6 months. The size of the aneurysm increased in 1 (2.0%) patient, was not changed in 6 (12.0%) patients, shrank in 34 (68.0%) patients, and was obliterated in 9 (18.0%) patients., Conclusions: The frozen elephant trunk for extensive aortic aneurysms had long-term durability and efficacy and might become the alternative treatment for extended aortic replacement., (Copyright 2010 The American Association for Thoracic Surgery. Published by Mosby, Inc. All rights reserved.)

A major cause of death in patients with bladder tumors is recurrence with metastasis. Bladder tumor metastasis is largely dependent upon the invasive capacity of tumor cells. Tumor cell invasion is mainly mediated by actin-rich protrusive membrane structures called invadopodia. The formation of invadopodia was observed in various types of invasive tumors such as breast cancer and melanomas. However, invadopodia formation so far has not been described in bladder tumor cells. We here report that human bladder tumor cells form functionally active invadopodia. By using a confocal laser scanning microscope, we demonstrated that invasive bladder tumor cell lines, YTS-1 and T24, with high Matrigel degradation activity form invadopodia but that noninvasive bladder tumor cell lines, RT4 and KK-47, form no detectable invadopodia. Invadopodia formed by YTS-1 cells had the ability to secrete matrix metalloproteases and degrade extracellular matrix to invade surrounding areas. Moreover, we observed that primary tumor cells obtained from patients with invasive bladder tumors also form invadopodia, validating the results from bladder tumor cell lines. Our results provide evidence that invasive human bladder tumor cells form invadopodia for tumor invasion.

We present a case of hepatoid carcinoma of the abdominal skin in a male Wistar rat. Histopathologically, this carcinoma resembled human hepatocellular carcinoma with respect to trabecular-sinusoidal structures. Carcinoma tissues contain numerous eosinophilic globules and crystals, and in this case, we found the characteristic eosinophilic globules in the hepatoid carcinoma cells and the crystals in the extracellular portions. Vivid carcinoma cells full of eosinophilic globules were present near the necrotic areas in tumor tissue, wherein quadrate crystals unstained with eosin were observed. PAS staining after diastase digestion revealed that the globules were PAS positive and diastase resistant. In addition, we found that the hepatoid carcinoma cells were immunoreactive for alpha-1-antitrypsin (anti-A1AT) antibody with the globules and crystals staining peripherally, and a central unstained region. Ultrastructural study of intracytoplasmic globules and extracellular crystals revealed that the fringe of each globule and crystal had no limiting membrane and showed the same level of electron density. These findings suggest that the characteristic crystals in this tumor may have originated from the globules that were emitted from the carcinoma cells after their death as a result of saturation with intracytoplasmic globules.

Background: This report compares long-term results with total arch replacement with frozen elephant trunk (FET) to ascending aortic or hemiarch replacement (AHR) for acute type A aortic dissection., Methods: The subjects were 120 consecutive patients, including 65 who received FET and 55 who had AHR for acute type A aortic dissection from 1997 to 2008. The late results after surgery were retrospectively compared between the FET and ARH groups., Results: Three patients in the FET group died, and 2 patients in the AHR group died. In long-term follow-up (mean, 67 months), the survival rate after 5 years was 95.3% for the FET group and 69.0% for the AHR group (p = 0.03). The event rate for the thoracic aorta after 5 years showed a significant difference between the FET and AHR groups (95.7% versus 73.0%, p = 0.01). A false lumen at the proximal descending aorta was patent in 16 patients (29%) in the AHR group, but it was thrombosed in all in the FET group., Conclusions: In patients with acute type A aortic dissection, it is possible to perform extensive primary repair using the FET technique with relative safety. FET may reduce the necessity for further operations to manage a residual false lumen.