WHOLE-BODY PROTEIN TURNOVER IN GOATS ENHANCED BY SUPPLEMENTING A DIET WITH RUMEN PROTECTED METHIONINE

Muramatsu, T.,Hatano, T.,Ueda, Y.,Furuse, M.,Okumura, J. Asian Australasian Association of Animal Productio 1994 Animal Bioscience Vol.7 No.2

Three experiments were conducted with female Japanese Saanen goats to investigate the effects of rumen protected methionine (RPMet) on N utilization and whole-body protein turnover. Whole-body leucine flux from which whole-body protein turnover rates were derived was measured by primed- continuous infusion of L-[$^{15}N$] leucine in combination with gas chromatography-mass spectrometry. Throughout the experiments RPMet was added to a diet to supply 1.5 g DL-methionine per goat per day. Irrespective of the major N sources (i.e., protein or urea) in the diet, both N deposition and whole-body protein synthesis were increased (p<0.05), and urinary N excretion was decreased (p<0.05) by supplementing with RPMet, but not by supplementing with methionine. It was concluded, therefore, that under the present experimental conditions, the RPMet supplement was efficiently bypassed to result in enhanced body protein synthesis of the goat.

EFFECT OF BACTERIAL INOCULATION ON NEUTRAL DETERGENT FIBRE DIGESTION AND ENERGY AVAILABILITY IN GERM-FREE CHICKENS

Muramatsu, T.,Niwa, N.,Furuse, M.,Okumura, J.,Ohmiya, K. Asian Australasian Association of Animal Productio 1992 Animal Bioscience Vol.5 No.1

The present study was done to examine whether inoculated and established bacteria in the digestive tract of germ-free (GF) chickens affect growth performance, energy availability, nitrogen utilization and neutral detergent fibre (NDF) digestibility of the host bird fed a high-fibre diet. Gnotobiotic (GB) chicks were made from GF birds by co-inoculating with Ruminococcus albus, and Staphylococcus warneri, only the latter of which was established in the chicken gut. No difference was detected among conventional (CV), GF and GB birds in body weight gain, food intake or food efficiency from 7 to 21 d of age. The amount of nitrogen retained was larger in CV than in GF and GB chicks. DE and ME values of the diet and NDF digestibility were higher in CV birds than in GF and GB counterparts. It was concluded, therefore, that the established bacterium S. warneri did not give any beneficial effects on the host bird as judged by growth performance, energy availability, nitrogen utilization, and NDF digestibility.

STUDIES ON METHIONINE METABOLISM IN THE RUMEN BACTERIA OF GOATS

Muramatsu, T.,Numa, M.,Ueda, Y.,Furuse, M.,Okumura, J.,Samukawa, K. Asian Australasian Association of Animal Productio 1994 Animal Bioscience Vol.7 No.2

The metabolic fate of methionine in rumen bacteria was studied by intraruminal administration of $^{15}N$ and $1-^{13}C$ labeled methionine in goats. Time course changes in isotopic abundance of amino acids in the rumen bacteria were determined with a computer-controlled gas-chromatograph mass spectrometer. The results from the transition of peak isotopic abundance in amino acids indicated that in rumen bacteria the $^{15}N$ or $^{13}C$ isotope in the methionine molecule was transferred rapidly to into bacteria, methionine administered intraruminally may not be retained as it is, but would be converted quickly to other metabolites in the bacteria.

EFFECTS OF LYSINE OR RUMINALLY PROTECTED LYSINE ADMINISTRATION ON NITROGEN UTILIZATION IN GOATS FED A DIET SUPPLEMENTED WITH RUMINALLY PROTECTED METHIONINE

Muramatsu, T.,Tsutsumi, K.,Hatano, T.,Hattori, M.,Okumura, J. Asian Australasian Association of Animal Productio 1993 Animal Bioscience Vol.6 No.3

The objectives of the present study were to investigate whether or not dietary lysine addition could improve N balance of female Japanese Saanen goats at 15 to 32 months of age, weighing 31 to 40 kg, fed on a wheat bran-hay cube diet supplemented with methionine, and whether or not ruminally protected lysine supplementation could give as good an N balance performance as lysine in the presence of ruminally protected methionine when given orally to the goats. It was considered from changes in N balance and N utilization that the first-and second-limiting amino acids in the diet were methionine and lysine respectively, under the present experimental conditions. The ruminally protected lysine in addition to the ruminally protected methionine gave no improvement in N balance and N utilization compared with the ruminally protected methionine alone, suggesting that the ruminally protected lysine used in the present study was not effectively utilized by the goats.

EFFECT OF RUMEN-PROTECTED METHIONINE ON WHOLE-BODY PROTEIN SYNTHESIS IN GOATS

Muramatsu, T.,Hatano, T.,Ueda, Y.,Furuse, M.,Okumura, J. Asian Australasian Association of Animal Productio 1989 Animal Bioscience Vol.2 No.3

COMPARISON OF UTILIZATION OF CELLULOSE AND CORN DIETARY FIBER AS AN ENERGY SOURCE IN CHICKS

Muramatsu, T.,Morishita, T.,Furuse, M.,Okumura, J. Asian Australasian Association of Animal Productio 1992 Animal Bioscience Vol.5 No.1

An experiment was conducted to investigate effects of fiber source on growth performance, N and neutral detergent fiber (NDF) digestibility, and utilization of energy in chicks fed an isocaloric low-energy diet from 7 to 21 days of age. Two fiber sources, cellulose and corn dietary fiber (CDF), were included in a diet at 10, 20 and 30% at the expense of kaolin, an inert diluent. The CDF contained 76.5% NDF consisting mainly of hemicellulose. The results showed that growth performance, N and NDF digestibility, dietary DE and ME values, energy deposition, and NE for production in birds fed CDF were inferior to those in birds fed cellulose. It can be concluded, from the present study, that chicks can utilize cellulose more efficiently than CDF up to a level of 30%.

Immunolocalization of laminin and integrin in regenerating junctional epithelium of mice after gingivectomy

Masaoka, T.,Hashimoto, S.,Kinumatsu, T.,Muramatsu, T.,Jung, H.-S.,Yamada, S.,Shimono, M. Blackwell Publishing Ltd 2009 Journal of periodontal research Vol.44 No.4

<P>Background and Objective: </P><P>The expression patterns of adhesive proteins and extracellular matrix proteins in regenerating gingival epithelium after gingivectomy are unknown. The aim of this study was to examine the expression of laminin 1, laminin &ggr;<SUB>2</SUB> (a specific component of laminin 5), integrin &bgr;<SUB>4</SUB> and integrin &agr;<SUB>3</SUB> in the regenerating gingival epithelium in order to understand the mechanism of wound healing during reconstitution of the sulcular environment.</P><P>Material and Methods: </P><P>The palatal gingivae of the maxillary molars of Institute of Cancer Research mice were excised, and the regenerating tissues were examined 1, 3, 5, 7 and 14 days later. Fresh, non-fixed and non-decalcified frozen sections were prepared and stained using immunofluorescence.</P><P>Results: </P><P>At 1 day post-surgery, intense expression of laminin &ggr;<SUB>2</SUB>, integrin &bgr;<SUB>4</SUB> and integrin &agr;<SUB>3</SUB> was distinct in the frontal margin of the regenerating oral epithelium. Laminin &ggr;<SUB>2</SUB> was diffusely detected on the root surface and in connective tissues beneath the regenerating oral epithelium at 3 and 5 days. At 7 days, laminin &ggr;<SUB>2</SUB> was intermittently recognizable in the internal basal lamina (IBL) close to tooth-facing cells, while laminin &ggr;<SUB>2</SUB>, integrin &bgr;<SUB>4</SUB> and integrin &agr;<SUB>3</SUB> were observed in the IBL and in the external basal lamina (EBL) of the regenerating junctional epithelium at 14 days.</P><P>Conclusion: </P><P>These results suggest that secretion of laminin 5 in the connective tissue may induce epithelial cell migration, and that binding of laminin 5 to integrin &agr;<SUB>6</SUB>&bgr;<SUB>4</SUB> and integrin &agr;<SUB>3</SUB>&bgr;<SUB>1</SUB> in the IBL may provoke cell adhesion and migration of cells facing the tooth on the enamel surface of the regenerating junctional epithelium.</P>

Involvement of laminin and integrins in adhesion and migration of junctional epithelium cells

Kinumatsu, T.,Hashimoto, S.,Muramatsu, T.,Sasaki, H.,Jung, H-S,Yamada, S.,Shimono, M. Blackwell Publishing Ltd 2009 Journal of periodontal research Vol.44 No.1

<P>Background and Objective: </P><P>The junctional epithelium attaches to the enamel surface with hemidesmosomes (of which laminin-5 and integrin-&agr;<SUB>6</SUB>&bgr;<SUB>4</SUB> are the main components) in the internal basal lamina. Laminin-5 is also involved in cell motility with integrin-&agr;<SUB>3</SUB>&bgr;<SUB>1</SUB>, although their functions have not yet been clarified. The purpose of this study was to determine the functions of those adhesive components between the tooth and the junctional epithelium during cell migration. Because an idea has been proposed that directly attached to tooth cells (DAT cells) may not contribute to cell migration, 5-bromo-2-deoxyuridine staining was performed to confirm cell migration.</P><P>Material and Methods: </P><P>We investigated laminin-&ggr;<SUB>2</SUB> (contained only in laminin-5), integrin-&bgr;<SUB>4</SUB> (involved in cell–extracellular matrix contact) and integrin-&agr;<SUB>3</SUB> (inducing cell migration) in the junctional epithelium, oral gingival epithelium and gingival sulcus epithelium of 6-wk-old ICR mice using laser microdissection, quantitative real-time reverse transcription-polymerase chain reaction, immunofluorescence and 5-bromo-2-deoxyuridine staining.</P><P>Results: </P><P>Laminin and integrins were clearly immunolocalized in the basal lamina of all epithelium. Quantitative analysis of laminin and integrin mRNAs by laser microdissection showed that they were more highly expressed in DAT cells than in basal cells in the oral gingival epithelium. In particular, a 12-fold higher expression of laminin-5 was observed in the junctional epithelium compared with the oral gingival epithelium. 5-Bromo-2-deoxyuridine staining showed rapid coronal migration of DAT cells.</P><P>Conclusion: </P><P>These results suggest that the abundant expression of laminin-5 and integrin-&agr;<SUB>6</SUB>&bgr;<SUB>4</SUB> is involved in the attachment of DAT cells to teeth by hemidesmosomes. Abundant expression of laminin-5 and integrin-&agr;<SUB>3</SUB>&bgr;<SUB>1</SUB> might assist in DAT cell migration, confirmed by 5-bromo-2-deoxyuridine staining during the turnover of junctional epithelium.</P>

Wide Area Distribution of Nitrogen Concentrations in Mountain Streams of Hyogo Prefecture, Japan

K. Muramatsu,Y. Komai,S. Umemoto,T. Inoue 대한환경공학회 2010 Environmental Engineering Research Vol.15 No.2

To study the relationship between the concentrations of nitrogen in mountain streams, and anthropologic and natural factors, the water chemistry of the mountain streams in the entire Hyogo Prefecture, Japan, were investigated. A thousand mountain streams were investigated between 1998 and 2001. The concentrations of nitrate nitrogen ranged from 2.92 to 0.1 mg/L, with an arithmetic mean value of 0.45 mg/L. A number of streams showing more than 1.0 mg/L of nitrate nitrogen accounted for 8% of the mountain streams investigated. These results indicated that the concentrations of nitrate nitrogen in the mountain streams were low in the entire Hyogo Prefecture. In general, the mountain stream water in Hyogo Prefecture appears to not have been affected by wet and dry deposition originating from anthropologic sources in mountain streams and Japan. On the other hand, sites with more than 0.8 mg/L nitrate nitrogen were distributed over the entire Hyogo Prefecture, which were classified into five groups. Each group showed unique geographical, geological and anthropological characteristics. No common characteristic among five groups were discover. These results suggest that the cause of high concentrations of nitrogen in mountain streams is not from a uniform set of conditions.

EARLY SCREENING OF EXPRESSION OF SV40 DRIVEN LACZ INTRODUCED INTO BOVINE EMBRYOS

Nakamura, A.,Okumura, J.,Muramatsu, T. Asian Australasian Association of Animal Productio 1995 Animal Bioscience Vol.8 No.5

The present study was conducted to assess gene expression of bacterial lacZ driven by the SV40 promoter at early developmental stages of bovine embryos. The lacZ gene was linearized with BamHI digestion and introduced into the pronucleus by microinjection at 20 hrs after the commencement of in vitro fertilization. Intact bovine blastocysts were not stained with X-Gal, suggesting that there is no endogenous beta-galactosidase activity in these blastocysts. In contrast, the bovine blastocyst cells microinjected with the lacZ gene exerted a characteristic greenish-blue color originating from the bacterial beta-galactosidase activity, albeit at a low rate, i.e. 2.1% of the total fertilized oocytes injected. It was concluded, therefore, that the lacZ gene driven by the SV40 promoter could be used for an indirect screening method in which the presence of transgene is evaluated from the product of transgene expression.