Thang, Tran Van;Sunagawa, Katsunori;Nagamine, Itsuki;Kishi, Tetsuya;Ogura, Go
Asian-Australasian Journal of Animal Sciences
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v.25
no.3
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pp.341-352
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2012
In large-type goats that were fed on dry forage twice daily, dry forage intake was markedly suppressed after 40 min of feeding had elapsed. The objective of this study was to determine whether or not marked decreases in dry forage intake after 40 min of feeding are mainly caused by the two factors, that is, ruminal distension and increased plasma osmolality induced thirst produced by dry forage feeding. Six large-type male esophageal- and ruminal-fistulated goats (crossbred Japanese Saanen/Nubian, aged 2 to 6 years, weighing $85.1{\pm}4.89kg$) were used in two experiments. The animals were fed ad libitum a diet of roughly crushed alfalfa hay cubes for 2 h from 10:00 to 12:00 am during two experiments. Water was withheld during feeding in both experiments but was available for a period of 30 min after completion of the 2 h feeding period. In experiment 1, saliva lost via the esophageal fistula was replenished by an intraruminal infusion of artificial parotid saliva (RIAPS) in sham feeding conditions (SFC) control, and the treatment was maintained under normal feeding conditions (NFC). In experiment 2, a RIAPS and non-insertion of a balloon (RIAPS-NB) control was conducted in the same manner as the SFC control of experiment 1. The intraruminal infusion of hypertonic solution and insertion of a balloon (RIHS-IB) treatment was carried out simultaneously to reproduce the effects of changing salt content and ruminal distension due to feed entering the rumen. The results of experiment 1 showed that due to the effects of multiple dry forage suppressing factors when feed boluses entered the rumen, eating rates in the NFC treatment decreased (p<0.05) after 40 min of feeding and cumulative dry forage intake for the 2 h feeding period reduced to 43.8% of the SFC control (p<0.01). The results of experiment 2 indicated that due to the two suppressing factors of ruminal distension and increased plasma osmolality induced thirst, eating rates in the RIHS-IB treatment were, as observed under NFC, reduced (p<0.05) and cumulative dry forage intake for the 2 h feeding period decreased to 34.0% of the RIAPS-NB control (p<0.01). The combined effects of ruminal distension and increased plasma osmolality accounted for 77.5% of the suppression of dry forage intake 40 min after the start of dry forage feeding. The results indicate that ruminal distension and increased plasma osmolality induced thirst are the main factors in the suppression of dry forage intake in large-type goats.
Thang, Tran Van;Sunagawa, Katsunori;Nagamine, Itsuki;Ogura, Go
Asian-Australasian Journal of Animal Sciences
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v.24
no.8
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pp.1069-1085
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2011
In large-type goats that were fed on dry forage twice daily, dry forage intake was markedly suppressed after 40 min of feeding had elapsed. The objective of this study was to clarify whether or not increases in plasma osmolality and subsequent thirst sensations produced by dry forage feeding suppress dry forage intake. Eight large-type male esophageal- and ruminal-fistulated goats (crossbred Japanese Saanen/Nubian, aged 3 to 6 years, weighing $72.3{\pm}2.74$ kg) were used in two experiments conducted under sham feeding conditions. The animals were fed ad libitum a diet of roughly crushed alfalfa hay cubes for 2 h from 10:00 to 12:00 h during two experiments. Water was withheld during feeding in both experiments but was available for a period of 30 min after completion of the 2 h feeding period. In experiment 1, an intraruminal infusion of artificial parotid saliva (RIAPS) in the control replenished saliva lost via the esophageal fistula and an intraruminal infusion of hypertonic solution (RIHS) in the treatment was carried out in order to reproduce the effects of changing salt content due to feed entering the rumen. In experiment 2, the RIHS control was conducted in the same manner as the RIHS treatment of experiment 1. The treatment group consisted of RIHS-with an intravenous infusion of artificial mixed saliva (VIAMS) treatment that was carried out for 3 h to prevent increases in plasma osmolality during feeding. The results of the RIHS treatment in experiment 1 showed that ruminal fluid osmolality increased and then an increase in plasma osmolality was observed. This resulted in the production of thirst sensations and the reduction of cumulative dry forage intake to 43.3% (p<0.05) of the RIAPS control. The results of the RIHS-VIAMS treatment in experiment 2 indicated that ruminal fluid osmolality was the same as the RIHS control but plasma osmolality significantly decreased, and thirst level was markedly reduced. This caused a significant increase of 31.4% (p<0.05) in cumulative dry forage intake in the RIHS-VIAMS treatment compared to the RIHS control. These results indicate that increases in ruminal fluid osmolality during dry forage feeding indirectly suppresses dry forage intake by causing an increase in plasma osmolality and subsequently inducing thirst sensations. The results of the present study suggest that marked decreases in dry forage intake after 40 min of feeding are caused by increases in plasma osmolality and subsequent thirst sensations produced by dry forage feeding.
Surface membrane proteins of virulent RH strain and tissue cyst-forming Fukaya strain of Toxoplasma gondii were analysed by SDS-polyacrylamide gel electrophoresis after LPO-catalyzed surface iodination and lectin blotting, then identified the zoite-specific antigens. Prior to the analyses, purification of RH tachyzoites from mouse peritoneal exudate and of Fukaya bradyzoites from mouse brain tissues were performed by centrifugation - on the discontinuous Percoll density-gradient. Ta- chysoites were obtained at the interface of 50U and 60% Percoll solution and brain cysts were harvested at the interfaces of 40-50% and 50-60%, then bradyzoites were obtained by treating the cysts with hypertonic solution. The LPO-catalyzed iodination detected 15 KDa and 14 KDa proteins o( brady- zoites and 30 KDa protein of tachysoites as major bands with several other minor bands. But Con A blotting revealed some bands of 200 K∼50 KDa glycoproteins of bradyzoites and 52 KDa band as major and minor bands of 33 K∼20 KDa of tachyzoites. Phytohemagglutinin did not detect any band in the two forms. EITB with anti- Fukaya antibody and anti-RH antibody revealed cross-reactivities between the two forms. Despite the cross-reactivity, anti-Fukaya antibody reacted with 15 KDa band of bradyzoites specifically and, anti-RH antibody with 52 KDa, 30 KDa, and 25 KDa bands of tachyzoites, respectively. It was identified that 15 KDa protein in bradyzoite, which was not a glycoprotein, was a major membrane protein with sufficient antigenicity, and in the case of tacky- zoite, 52 KDa surface glycoprotein (gp52) with specific antigenicity might be added to the major surface protein, p30.
Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disorder that causes progressive paralysis. L-Citrulline is a nonessential neutral amino acid produced by L-arginine via nitric oxide synthase (NOS). According to previous studies, the pathogenesis of ALS entails glutamate toxicity, oxidative stress, protein misfolding, and neurofilament disruption. In addition, L-citrulline prevents neuronal cell death in brain ischemia; therefore, we investigated the change in the transport of L-citrulline under various pathological conditions in a cell line model of ALS. We examined the uptake of [14C]L-citrulline in wild-type (hSOD1wt/WT) and mutant NSC-34/ SOD1G93A (MT) cell lines. The cell viability was determined via MTT assay. A transport study was performed to determine the uptake of [14C]L-citrulline. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was performed to determine the expression levels of rat large neutral amino acid transported 1 (rLAT1) in ALS cell lines. Nitric oxide (NO) assay was performed using Griess reagent. L-Citrulline had a restorative effect on glutamate induced cell death, and increased [14C]L-citrulline uptake and mRNA levels of the large neutral amino acid transporter (LAT1) in the glutamate-treated ALS disease model (MT). NO levels increased significantly when MT cells were pretreated with glutamate for 24 h and restored by co-treatment with L-citrulline. Co-treatment of MT cells with L-arginine, an NO donor, increased NO levels. NSC-34 cells exposed to high glucose conditions showed a significant increase in [14C]L-citrulline uptake and LAT1 mRNA expression levels, which were restored to normal levels upon co-treatment with unlabeled L-citrulline. In contrast, exposure of the MT cell line to tumor necrosis factor alpha, lipopolysaccharides, and hypertonic condition decreased the uptake significantly which was restored to the normal level by co-treating with unlabeled L-citrulline. L-Citrulline can restore NO levels and cellular uptake in ALS-affected cells with glutamate cytotoxicity, pro-inflammatory cytokines, or other pathological states, suggesting that L-citrulline supplementation in ALS may play a key role in providing neuroprotection.
Rodrigo Romero Correa;Rubens Peres Mendes;Diego Darley Velasquez Pineros;Aymara Eduarda De Lima;Andre Luis do Valle De Zoppa;Luis Claudio Lopes Correia da Silva;Ricardo de Francisco Strefezzi;Silvio Henrique de Freitas
Journal of Veterinary Science
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v.25
no.2
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pp.29.1-29.11
/
2024
Background: Preservation of biological tissues has been used since ancient times. Regardless of the method employed, tissue preservation is thought to be a vital step in veterinary surgery teaching and learning. Objectives: This study was designed to determine the usability of chemically preserved cadaveric equine heads for surgical teaching in veterinary medicine. Methods: Six cadaveric equine heads were collected immediately after death or euthanasia and frozen until fixation. Fixation was achieved by using a hypertonic solution consisting of sodium chloride, sodium nitrite and sodium nitrate, and an alcoholic solution containing ethanol and glycerin. Chemically preserved specimens were stored at low temperatures (2℃ to 6℃) in a conventional refrigerator. The specimens were submitted to gross and organoleptic assessment right after fixative solution injection (D0) and within 10, 20, and 30 days of fixation (D10, D20, and D30, respectively). Samples of tissue from skin, tongue, oral vestibule, and masseter muscle were collected for histological evaluation at the same time points. Results: Physical and organoleptic assessments revealed excellent specimen quality (mean scores higher than 4 on a 5-point scale) in most cases. In some specimens, lower scores (3) were assigned to the range of mouth opening, particularly on D0 and D10. A reduced the range of mouth opening may be a limiting factor in teaching activities involving structures located in the oral cavity. Conclusions: The excellent physical, histologic, and organoleptic characteristics of the specimens in this sample support their usability in teaching within the time frame considered. Appropriate physical and organoleptic characteristics (color, texture, odor, and flexibility) of the specimens in this study support the use of the method described for preparation of reusable anatomical specimens.
Kim, Ki-Up;Kim, Yang-Ki;Shin, Chan-Young;Kim, Do-Jin;Uh, Soo-Taek;Kim, Yong-Hoon;Ko, Kwang-Ho;Park, Choon-Sik
Tuberculosis and Respiratory Diseases
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v.49
no.1
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pp.82-92
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2000
Background : In chronic airway disease, mucus secretion is increased, but extraction of mucin, which is the main component of mucus secretion, is a very complicated and limited in clinical use. Recently, monoclonal antibody for mucin was developed for possible clinical use. In this study, cellular analysis and quantification of respiratory mucin in sputum of patients with chronic airway diseases were performed. Method : Sputum was collected from patients with asthma(n=33), bronchiectasis(n=8) or chronic bronchitis (n=13) by spontaneous expectoration or by hypertonic saline induction. Collected sputums was treated by 0.1% dithiotreitol to dissociate the disulfide bond of the mucus and filtered through a nylon gauze. Total cell count, viability and differential count were measured. For detection of mucin, collected samples were treated with sodium dodecyl sulfate polyacrylamide gel electrophoresis and then with monoclonal antibody(HMO2), as the primary antibody, and PAS stain. The amount of mucin was measured with ELISA by HMO2. Correlation with clinical information, cellular analysis, and amount of measured mucin were analyzed. Results : Total cell counts of sputum were significantly increased in patients with bronchiectasis but viability remained the same. Eosinophils were significantly increased in patients with asthma, neutrophils in bronchiectasis chronic bronchitis, respectively (p<0.05). The results of Western blotting and PAS staining confirmed the presence of glycoproteins and matched? with mucin. The amounts of mucin measured by ELISA were not significantly different among the disease groups. Significant correlation was identified between the amount of mucin and viability(r=-0.482, p<0.05). Conclusion : Inflammatory cells in the sputum of those with chronic airway disease were different for each disease type. Measurement of mucin by ELISA via monoclonal antibodies may be a simple method for the evaluation of chronic airway disease.
Park, Young-Hoon;Ahn, Soo-Ho;Shin, Son-Moon;Hah, Jeong-Ok
Journal of Yeungnam Medical Science
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v.8
no.2
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pp.128-137
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1991
Peritoneal dialysis has been widely considered to be the dialytic treatment of choice for acute renal failure in infants and young children, because the technique is simple, safe and easily adapted for these patients. Also peritoneal dialysis in infants might have more effective ultrafiltration and clearance than in adults. In certain circumstances associated with hemodynamic instability, ordinary volume peritoneal dialysis(30-50 ml/kg body weight per exchange) or hemodialysis may not be suitable unfortunately. But frequent cycled, low volume, high concentration peritoneal dialysis may be more available to manage the hemodynamically untable acute renal failure of newborns and infants. Seven infants underwent peritoneal dialysis for hemodynamically unstable acute renal failure with low exchange volume($14.2{\pm}4.2ml/kg$), short exchange time(30 to 45 minutes) and hypertonic glucose solution(4.25% dextrose). Age was $1.9{\pm}1.3$ months and body weight was $4.6{\pm}1.6kg $. Etiology of acute renal failure was secondary to sepsis with or without shock(5 cases) and postcardiac operation(2 cases). Catheter was inserted percutaneously with pigtail catheter or Tenkhoff catheter by Seldinger method. Dialysate was commercially obtained Peritosol which contained sodium, chloride, potassium, magnesium, lactate and calcium. Net ultrafiltration(ml/min) showed no difference between low volume dialysis and control($0.27{\pm}0.09$ versus $0.29{\pm}0.09$) Blood BUN decreased from $95.7{\pm}37.5$ to $75.7{\pm}25.9mg/dl$ and blood pH increased from $7.122{\pm}0.048$ to $7.326{\pm}0.063$ after 24 hours of peritoneal dialysis. We experienced hyperglycemia which were controlled by insulin(2 episodes), leakage at the exit site(2), mild hyponatremia(1) and Escherichia coli peritonitis(1). Two children of low volume dialysis died despite the treatment. In our experience, low volume and high concentration peritoneal dialysis with frequent exchange may have sufficient ultrafiltration and clearance without significant complications in the certain risked acute renal failure of infants.
Background: Bioprosthetic materials have been made using glutaraldehyde fixation of porcine or bovine pericardium during cardiovascular surgery. But these bioprostheses have the problems of calcification and mechanical failure. We determined changes in tensile strength and elasticity of pericardium after glutaraldehyde, solvent, decellularization and detoxification. Material and Method: Tissues were allocated to four groups: glutaraldehyde with and without solvent, decellularization, and detoxification. We studied tensile strength and strain on tissues. We measured the tensile strength of fresh pericardium stretched in six directions (with 5 mm width), and % strain, which we calculated from the breaking point when we pulled the pericardium in two directions. Result: Tensile strength was reduced when we used the usual concentrated glutaraldehyde fixation (n=83, $MPa=11.47{\pm}5.40$, p=0.006), but there was no change when we used solvent. Elasticity was increased after glutaraldehyde fixation (n=83, strain $(%)=24.55{\pm}9.81$, p=0.00), but there was no change after solvent. After decellularization of pericardium, the tensile strength was generally reduced. The decrease in tensile strength after concentrated glutaraldehyde fixation for a long time was significantly greater less than after concentrated solvent (p=0.01, p=0.00). After detoxification, the differences in strength and strain were not significant. Conclusion: After glutaraldehyde treatment of pericardium there is no loss in tensile strength (even though we did the glutaraldehyde, solvent and detoxification treatments LOGIC IS UNCLEAR). Also, these treatments had a tendency to increase elasticity. Although post-treatment decellularization led to a significant loss in strength, this effect could be attenuated using a low concentration of solvent or hypertonic solution.
The rice variety, Kwanok, was reared in the water, land and salty beds and transplanted to the reclaimed soil area having an average of 0.48% salt concentration 0.67% at the end of April Two levels of $NH_3$-N and urea-N with 6 treatments were used. The effect of each treatment was observed. The plant height of the land bed seedlings at transplanting stage was short but the dry-weight/plant-height ratio was large, and the rooting ability vigorous remarkably after transplantation in the salty area. The total carbohydrate content of the stem part was markedly larger in the land bed seedlings than the others and the C/N ratio was accordingly greater. This tendency was observed through the last rooting stage. In the salty nutrient solution, the roots of salty bed seedlings showed high respiratory activity. The activity of the land bed seedlings did not decreased in the hypertonic solution as much as the water bed seedlings. There was no difference in the effect of fertilization on the rough rice yield between ammonium sulphate and urea. The cultural practices with the land and salty bed seedlings increased the rough rice yields by 33% and 22% respectively, compared with the yields of the water bed Seedlings. The number of panicles, panicle weight and the number of grains per panicle were much greater from the rice plant grown by the land bed seedlings than from the other bed seedlings.
Background : It has been well known that bronchia1 asthma is a chronic airway inflammatory disorder. Recently, sputum induced with hypertonic saline was introduced as a simple and useful nonivasive medium to investigate airway inflammation and symptom severity in patients with asthma. We examined the eosinophil, eosinophil cationic protein (ECP), interleukin(IL)-3, IL-5, granulocyte-macrophage colony-stimulating facta (GM-CSF), and nitric oxide (NO) derivatives in induced sputum from patients with bronchia1 asthma in order to determine the role of NO and various inflammatory cytokines as a useful markers of airway inflammation or changes in pulmonary function tests and symptoms. Methods : A total 30 patients with bronchia1 asthma received oral prednisolone 30 mg daily for 2 weeks. Forced expiratory volume in one second ($FEV_1$), total blood eosinophil count and induced sputum eosinophil count, ECP, IL-3, IL-5, GM-CSF, and NO derivatives were determined before and after the administration of prednisolone. Results : Of the 30 patients, 13 (43.3%) were male and 17 (56.7%) were female. The mean age of patients was 41.8 years (range 19-64 years). Two patients could not produce sputum at the second study and 3 could not be followed up after their first visit. Two weeks after the prednisolone administration, there was a significant increase in $FEV_1$ (% of predicted value) from 78.1$\pm$20.6 % to 90.3$\pm$ 18.3 % (P<0.001). The eosinophil percentages in induced sputum were significantly decreased after treatment with prednisolone, with values of 56.1$\pm$27.2 % versus 29.6$\pm$21.3 % (P<0.001), and ECP were $134.5\pm68.1\;{\mu}g/L$ versus $41.5\pm42.4\;{\mu}g/L$ (P<0.001) respectively. After the prednisolone treatments, the eotaxin concentration also showed a decreasing tendency from 26.7$\pm$12.8 pg/ml to 21.7$\pm$8.7 pg/ml. There was a decreasing tendency but no significant differences in total blood eosinophil count (425.7$\pm$265.9 vs 287.7$\pm$294.7) and in the concentration of NO derivatives ($70.4\pm44.6{\mu}mol/L$ vs $91.5\pm48.3\;{\mu}mol/L$) after the prednisolone treatments. IL-3, IL-5, GM-CSF were undetectable in the sputum of most subjects either before the prednisolone treatments or after the treatments. Before the prednisolone treatments, a significant inverse correlation was observed between FEV1 and sputum ECP (r=-D.364, P<0.05) and there was a significant correlation between sputum eosinophils and eotaxin (r=0.369, P<0.05) Conclusion : The eotaxin and ECP concentration in induced sputum may be used as markers of airway inflammation after treatments in bronchia1 asthma. In addition, the measurement of sputum eosinophil percent ages is believed to be a simple method displaying the degree of airway inflammation and airway obstruction before and after the prednisolone treatment in bronchia1 asthma. However, unlike exhaled NO, the examination of NO derivatives with Griess reaction in induced sputum is considered an ineffective marker of changing airway inflammation and obstructing symptoms.
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