bluuebunny/crossref_metadata_2025_split huggingface dataset:
Twenty‐eight new microsatellite loci in chicken and their cross‐species amplification in Japanese quail and helmeted guinea fowl
Authors: KAYANG Boniface Baboreka , INOUE‐MURAYAMA Miho , TAKAHASHI Hideaki , MINEZAWA Mitsuru , TSUDZUKI Masaoki , MIZUTANI Makoto , ITO Shin’ichi 
Published: July 2003

ABSTRACTTwenty‐eight original chicken microsatellite markers were isolated and characterized to determine their utility as cross‐reactive markers for comparative genetic mapping in the order Galliformes. Primer pairs were typed in 12 unrelated chickens and also tested on Japanese quail and helmeted guinea fowl deoxyribonucleic acid (DNA). Polymorphism was observed in 23 (82.1%) of the markers and the average number of alleles per locus was 2.9 while the mean heterozygosity was 0.19. Eleven (39.3%) of the chicken markers cross‐reacted with Japanese quail DNA and 2 (7.1%) with helmeted guinea fowl DNA. The cross‐reactive markers described would serve as useful resources for comparative genetic mapping in poultry species belonging to the order Galliformes.
Development of transferred xenogeneic vole embryos in mouse uteri
Authors: WIDAYATI Diah Tri , OHMORI Yasushige , WAKITA Tsuyoshi , FUKUTA Katsuhiro 
Published: July 2003

ABSTRACTAn experimental model to study interspecific pregnancy using voles,Microtus arvalis, and green fluorescent protein gene‐induced transgenic mice is presented. Xenogeneic blastocysts from the vole were transferred into the uteri of pseudopregnant mice along with allogeneic blastocysts from green fluorescent protein gene‐induced transgenic mice. The uteri containing xeno‐allo combined transfers were examined from day 6 to 13 of gestation. Although the vole embryos implanted, the uteri containing vole embryos were smaller compared with those having allogeneic mouse embryos. On day 8, the uteri containing vole embryos hemorrhaged internally and no vole embryo was found in the pregnant uterus after day 11. Allogeneic mouse embryos developed normally despite the presence and abortion of the vole embryos. In uteri implanted with vole embryos, decidua were formed and numerous blood vessels were distributed around the embryo. Maternal blood cells infiltrated into the celomic cavity of the vole embryo through the discontinuous region of trophoblast. Periodic acid‐Schiff‐positive granulated metrial gland cells were remarkably increased in the decidual sites. These findings suggest that a disorder of embryo–maternal interaction might induce the appearance of numerous granulated metrial gland cells and rejection of the embryos.
Caprine somatic cell nuclear transfer using in vivo matured oocytes collected by laparoscopic follicular aspiration
Authors: OHKOSHI Katsuhiro , TAKAHASHI Seiya , KOYAMA Shin‐ichiro , AKAGI Satoshi , ADACHI Noritaka , FURUSAWA Tadashi , FUJIMOTO Jun‐ichiro , IZAIKE Yoshiaki , TOKUNAGA Tomoyuki 
Published: July 2003

ABSTRACT In vivo matured oocytes collected by laparoscopic follicular aspiration (LFA) from hormone treated female goats were used as recipient ooplasts for somatic cell nuclear transfer (SCNT). Japanese native (Shiba) goats were used as donor females and some donor females were used repeatedly (two or three times) at intervals of a few months. To induce synchronization of estrus, a sponge containing 0.5 g of progesterone was inserted into the vagina of each goat for 14 days. These animals were also treated with follicle stimulating hormone (FSH) in a series of 8 injections over 4 days. The first FSH injection was administered on the morning of day 9 of sponge insertion. On the morning of day 13, 50 µg of gonadotropin‐releasing hormone (GnRH) was injected into each animal. Twenty‐nine hours after GnRH injection, LFA was performed. After removal of cumulus cells, collected oocytes with the first polar body were selected and enucleated for nuclear transfer. Anterior pituitary cells isolated from an adult male Shiba goat were transfected with a DNA fragment containing the enhanced green flourescent protein gene and the puromycin resistance gene. A single donor cell was inserted into the perivitelline space of each enucleated oocyte and fusion was induced with one electric pulse of 20 V for 10 µs. The SCNT goat eggs were cultured in chemically defined medium at 38.5°C in 5% CO2, 5% O2, 90% N2 for 9 days. By LFA, 396 oocytes were collected from a total of 30 females. After removal of cumulus cells, 64% of them extruded the first polar body. The percentage of SCNT goat eggs produced using in vivo matured oocytes which developed to the blastocyst stage (20–21%) was significantly higher (P < 0.05) than that produced with in vitro matured oocytes (3–8%).
Molecular cloning of insulin‐like growth factor‐I complimentary DNA and promoter region in the domestic duck (Anas platyrhynchos)*
Authors: KANSAKU Norio , YAGI Eriko , NAKADA Asako , OKABAYASHI Hisato , GUEMENE Daniel , ZADWORNY David 
Published: July 2003

ABSTRACTComplementary DNA (cDNA) and the flanking region of insulin‐like growth factor‐I (IGF‐I) of domesticated duck were cloned. The nucleotide sequence analysis of the cDNA showed seven and eight bases different, respectively, from chicken and turkey IGF‐I cDNA within the coding region. The amino acid sequence of prepro IGF‐I differed by one and two amino acids from those observed in chicken and turkey, respectively. However, no amino acid substitution was observed in the mature IGF‐I region. Sequence analysis of the promoter region and exon 1 of the duck IGF‐I revealed a high degree of similarity to that of the chicken IGF‐I gene. These results suggest that the mechanisms which regulate expression of the IGF‐I gene may be widely conserved in avian species.
Synthesis of citrulline from ornithine by the small intestinal mucosa of cattle
Authors: SULTANA Halima , KITANO Ayumi , WADUD Shaila , TAKAHASHI Toshihiro , MORITA Tetsuo , ONODERA Ryoji 
Published: July 2003

ABSTRACTThree segments of cattle small intestine (duodenum, upper jejuno‐ileum and lower jejuno‐ileum) were examined in an in vitro system for activity of ornithine carbamoyltransferase (OCT; EC 2.1.3.3) which is involved in the synthesis of citrulline (Cit) from ornithine (Orn). The mucosa of the three segments of small intestine was collected from Japanese black cattle, homogenized and then centrifuged. The supernatant fraction was used as the crude OCT enzyme solution. The OCT activity was assayed by the production of Cit from Orn determined directly by HPLC. The optimal pH and temperature for OCT activities in the duodenum, upper jejuno‐ileum and lower jejuno‐ileum of cattle small intestine were 7.47 and 39°C. Little difference was observed between the three segments. The OCT activity in cattle kidney was also examined for comparison, and almost no activity was found. The OCT activities in crude enzyme solutions of the three segments of small intestine were stable for up to one month of storage at −20°C in Tris HCl buffer solution. Finally, the role of the small intestine in supplying Cit as a precursor for arginine synthesis in cattle kidney was discussed.
A quantitative study on arginine synthesis from argininosuccinic acid and citrulline by crude enzymes of cattle kidney
Authors: SULTANA Halima , INADA Mikiko , WADUD Shaila , TAKAHASHI Toshihiro , MORITA Tetsuo , ONODERA Ryoji 
Published: July 2003

ABSTRACT In vitro experiments were conducted to examine the synthesis of arginine (Arg) from argininosuccinic acid (ASA) and citrulline (Cit) by crude enzymes of cattle kidney cortex. Kidney samples, collected from Japanese black cattle, were homogenized in KCl solution (ice‐cold), and centrifuged at 27 000 × g for 20 min at 4°C, and the supernatant fluid was used as a crude enzyme solution. The enzyme solution was incubated at 39°C in Tris HCl buffer with 15 mmol/L ASA or with 10 mmol/L Cit in the presence of 10 mmol/L aspartic acid (Asp), 10 mmol/L ATP and 5 mmol/L MgCl2 to examine the activities of two enzymes, argininosuccinate lyase and argininosuccinate synthetase, which work at the terminal steps of Arg biosynthesis. The production of Arg from ASA, or ASA and Arg from Cit by argininosuccinate lyase and argininosuccinate synthetase activities, respectively, were determined directly by the HPLC method. The optimum pH for argininosuccinate lyase activity was 7.85. Unfortunately, the optimum pH for argininosuccinate synthetase activity could not be determined because no inhibitor of argininosuccinate lyase was used in the Cit incubation, so the ASA produced from Cit spontaneously converted to Arg during incubation with Cit. The maximum production of ASA from Cit was found at pH 6.45 under these conditions. We observed the optimum pH for the synthesis of Arg from Cit at 7.7. The production of Arg from ASA or Cit was quantitatively determined as 14.4 or 8.83 µmol/g kidney tissue/h, respectively, at the optimal pH values. This suggests that the daily production of Arg from ASA or Cit by the kidney might be sufficient to cover the daily requirement of Arg in cattle.
In vitro metabolism of tyrosine by rumen bacteria, protozoa and their mixture
Authors: KHAN Rokibul Islam , ANDO Sada , TAKAHASHI Toshihiro , MORITA Tetsuo , ONODERA Ryoji 
Published: July 2003

ABSTRACTAn anaerobic in vitro study of tyrosine (Tyr) metabolism by rumen bacteria (B), protozoa (P) and their mixture (BP) was conducted using an HPLC analytical method to obtain some basic and systematic information on aromatic amino acid metabolism in the rumen. Most of the Tyr (> 96%) disappeared after 12 h incubation in B and BP, but only 58% in P. Tyr was converted mainly to p‐hydroxyphenylacetic acid (HPA) in all microbial suspensions. About 45% of disappeared Tyr in B and P, and about 35% in BP were converted to HPA. An appreciable amount of phenylalanine (Phe), 13 and 3% of disappeared Tyr, and a small amount of tryptophan (Trp), 8 and 1% of disappeared Tyr, were also produced from Tyr by rumen bacteria and protozoa, respectively. Small amounts of p‐hydroxyphenylpyruvic acid (about 4 and 6% of disappeared Tyr) were produced from Tyr in B and P, respectively. A moderately large amount of phenylacetic acid (14% of disappeared Tyr) was produced from Tyr in P which was 1.9 times higher than that in B. Phenylpropionic acid and p‐hydroxybenzoic acid were produced only in B and BP. It was concluded that the Tyr degradation ability of rumen bacteria was about 1.5 times higher than that of rumen protozoa. Degraded Tyr mainly produces HPA and then two other aromatic amino acids, Phe and Trp, which are considered essential amino acids for ruminants. Therefore, it is speculated that the requirement for Phe and Trp in ruminants may be partially fulfilled if Tyr is sufficiently supplied in rations.
Effects of supplemental protein sources during the grazing period on performance, ruminal characteristics and blood constituents in early weaned growing Wagyu steer calves
Authors: SUGIMOTO Masahito , YAYOTA Cizuru , KUZUOKA Syuji , SATO Yukinobu 
Published: July 2003

ABSTRACTWe investigated the effects of grazing and supplemental protein sources during the grazing period on growth performance, ruminal characteristics and blood constituents in 15 growing Wagyu steer calves weaned at 3 months of age. Each animal was allotted one of three treatments: (1) GSBM, soybean meal (SBM) supplemented as a protein source during a grazing period; (2) GCGM, corn gluten meal (CGM) supplemented as a protein source during a grazing period; or (3) NG, no grazing, in which steers were fed hay and GSBM concentrate only. The estimated rumen undegradable protein (RUP) concentration of CGM was higher than that of SBM. High RUP supplementation during the grazing period did not influence feed intake or growth performance. Compared with the NG treatment, the average daily gain in GSBM and GCGM calves was significantly depressed (P < 0.05). Bodyweight/withers height ratio in the NG calves tended to be greater than in the GSBM and GCGM calves. Final paunch girth/heart girth ratio in the GSBM and GCGM calves was significantly greater (P < 0.05) than in the NG calves. Supplementation with CGM did not influence ruminal ammonia nitrogen and blood constituents. These results indicate that CGM supplementation during the grazing period did not enhance the growth of Wagyu calves because the CGM diet probably did not elevate the metabolizable protein supply to the grazing calves above that of the SBM diet in our conditions.
Production of aromatic amino acids and related compounds from p‐hydroxyphenylacetic acid by rumen microorganisms in vitro
Authors: KHAN Rokibul Islam , ANDO Sada , TAKAHASHI Toshihiro , MORITA Tetsuo , ONODERA Ryoji 
Published: July 2003

ABSTRACTSuspensions of mixed rumen bacteria (B), protozoa (P), and mixed rumen microorganisms (BP) prepared from rumen contents of fistulated goats were anaerobically incubated with 1 mM p‐hydroxyphenylacetic acid (HPA) at 39°C for 24 h. Tyrosine (Tyr), phenylalanine (Phe), tryptophan (Trp) and other related compounds in both supernatants and hydrolyzates of microbial cells in all incubations were analyzed by HPLC. Large amounts of Tyr (32.1, 42.7 and 36.1% of disappeared HPA in B, P and BP, respectively) were produced from HPA during a 12 h incubation period. The formation of Tyr in P (178.6 µmol/g MN) was 1.5 and 2 times higher than in B and BP, respectively. Phe (7–11% of the disappeared HPA) and Trp (3–6% of the disappeared HPA) were also synthesized from HPA in B, P, and BP. Phe synthesis in P (46.3 µmol/g MN) was 1.7 times higher than in B but, in contrast, Trp synthesis in B, was 1.6 times higher than in P. The metabolites p‐hydroxyphenylpyruvic acid (in the range of 5–14% of disappeared HPA), phenylacetic acid (1–11%), p‐hydroxybenzoic acid (3–7%) and benzoic acid (1–6%) were produced from HPA in B, P and BP. Phenylpropionic acid (6% of the disappeared HPA) was produced only in B and BP.
Effect of α‐cyclodextrin‐allyl isothiocyanate on ruminal microbial methane production in vitro
Authors: LILA Zeenat Ara , MOHAMMED Nazimuddin , KANDA Shuhei , KAMADA Toshihiko , ITABASHI Hisao 
Published: July 2003

ABSTRACTThe objective of the present study was to investigate the effects of α‐cyclodextrin‐allyl isothiocyanate (CD‐AI) on ruminal microbial methane production and rumen fermentation of corn starch, soluble potato starch or hay plus concentrate (1.5:1) by mixed rumen microorganisms. Diluted rumen fluid (30 mL) was incubated anaerobically at 38°C for 6 and 24 h with or without CD‐AI (0, 0.4, 0.8, 1.6 and 3.2 g/L). The pH of the medium was unchanged by CD‐AI in all substrates. The molar proportion of acetate was decreased and propionate was increased with a corresponding decrease in acetate : propionate ratio (P < 0.05). Total volatile fatty acids and butyrate were increased (P < 0.05). Ammonia‐N was decreased (P < 0.05). Except with soluble potato starch, numbers of protozoa were unchanged after 6 h. As concentration of CD‐AI increased from 0 to 3.2 g/L, fermentation of corn starch, soluble potato starch and hay plus concentrate resulted in decreased (P < 0.05) methane production of 49–100% (6 h) and 14–100% (24 h); 39–100% (6 h) and 16–100% (24 h); and 45–100% (6 h) and 17–100% (24 h), respectively. When hay plus concentrate was used as substrate, methanogenic bacteria were decreased (P < 0.05) with 0.8 g/L of CD‐AI after 6 h. Excluding the lower dose level (0.4 g/L) of CD‐AI, digestibility of neutral detergent fiber of hay plus concentrate was decreased (P < 0.05) after 24 h. A suitable level of CD‐AI could therefore be used as a supplement to inhibit methane production and improve rumen fermentation without detrimental effects on fiber digestion.
Fine needle aspiration (FNA) cyt_logy in tuberculous lymphadenitis
Authors: ERSÖZ  , POLAT  , SERIN  , SOYLU  , DEMIRCAN  
Published: July 2003

ersöz c., polat a., serin m. s., soylu l. and demircan o. (1998) Cyt_pathology9, 201–207Fine needle aspiration (FNA) cyt_logy in tuberculous lymphadenitisSixty‐three lymph node aspirates were screened and 32 aspirates revealing granulomatous lymphadenitis with or without caseation necrosis were re‐evaluated. The most characteristic morphological features among these cases were epithelioid cell clusters with or without caseation necrosis. When clusters were thick, careful observation of the periphery of the clusters helped to find epithelioid cells. Caseation necrosis revealed a typical macroscopic and microscopic appearance. Ziehl–Neelsen staining was negative in all smears and histological sections. Polymerase chain reaction (PCR) amplification technique was applied to 23 of the cases in which the cyt_logical diagnoses were consistent with tuberculosis. Mycobacterium tuberculosis was demonstrated in 19 (82.60%) cases. In conclusion: (i) it is necessary to perform several aspirations from different sites of the enlarged lymph node; (ii) the diagnosis of ‘granulomatous lymphadenitis, consistent with tuberculosis’ can be given, even though the acid‐fast stains are negative; (iii) additional techniques such as PCR give supportive information; (iv) an open biopsy is recommended if there is a discrepancy with the clinical impression.
The phosphatidylinositol‐3 kinase (PI3K)‐Akt pathway suppresses neurite branch formation in NGF‐treated PC12 cells
Authors: Higuchi Maiko , Onishi Keisuke , Masuyama Norihisa , Gotoh Yukiko 
Published: July 2003

AbstractBackground:  Previous studies have shown that phosphatidylinositol‐3 kinase (PI3K) plays an important role in NGF (nerve growth factor)‐induced neurite elongation. However, the roles of the PI3K pathway in neurite branch formation were not fully understood. Also, it was not clear where the PI3K pathway is activated during branch formation.Results:  We found that the treatment of PC12 cells with the PI3K inhibitor LY294002 resulted in a marked increase in the number of neurite branch points, suggesting a suppressive role of PI3K in neurite branch formation. Expression of a constitutively active form of Akt, a downstream effector of PI3K, decreased the number of branch points, whereas that of a dominant‐negative form of Akt increased it. In contrast, inhibition of neither Rac, mTOR nor GSK3, other effectors of PI3K, promoted branch formation. Importantly, the phosphorylated form of endogenous Akt was localized at the tips of growth cones, but devoid of small branches in NGF‐treated PC12 cells. A GFP‐fusion protein of the plekstrin‐homology (PH) domain of Akt was also localized at the tips of growth cones.Conclusions:  The PI3K‐Akt pathway thus plays a key role in suppression of neurite branch formation in NGF‐treated PC12 cells. working model for the regulation of neuritogenesis in PC12 cells. PI3K may mediate NGF regulation of neuritogenesis via two pathways. Rac induces neurite elongation and branch formation. Akt induces neurite elongation, but prevents branch formation.image
A tumour‐associated DEAD‐box protein, rck/p54 exhibits RNA unwinding activity toward c‐myc RNAs in vitro
Authors: Akao Yukihiro , Yoshida Hitoshi , Matsumoto Kenji , Matsui Tsutomu , Hogetu Keita , Tanaka Nobuo , Usukura Jiro 
Published: July 2003

AbstractBackground:  The rck/p54 protein of 473 amino acids belongs to the family of DEAD‐box/putative RNA helicase proteins. DEAD‐box proteins have been implicated in a wide variety of cellular processes ranging from the initiation of protein synthesis and ribosome biosynthesis to premRNA splicing by means of modifying the RNA structure. Our previous data suggested that rck/p54 positively affected the translation initiation of c‐myc mRNA.Results:  The data obtained from morphological studies and surface plasmon resonance assays clearly indicated that the protein specifically bound to c‐myc RNA transcripts (RNAs) and exhibited RNA unwinding activity toward c‐myc RNAs in the presence of ATP in vitro. Experiments using a deletion mutant of rck/p54 retaining only its N‐terminal 289 amino acids demonstrated that the deleted C‐terminal 184 amino acid domain is involved in the RNA unwinding activity.Conclusion:  These findings strongly suggest that rck/p54 may play an important role in translation initiation by restructuring mRNAs even in the cell and contribute to carcinogenesis.
Negative regulation of Wnt signalling by HMG2L1, a novel NLK‐binding protein
Authors: Yamada Misato , Ohkawara Bisei , Ichimura Naoya , Hyodo‐Miura Junko , Urushiyama Seiichi , Shirakabe Kyoko , Shibuya Hiroshi 
Published: July 2003

AbstractBackground:  Wnt signalling plays a critical role in many developmental processes and tumorigenesis. Wnt/β‐catenin signalling induces the stabilization of cytosolic β‐catenin, which interacts with TCF/LEF‐1 transcription factors, thereby inducing expression of Wnt‐target genes. Recent evidence suggests that a specific MAP kinase pathway involving the MAP kinase kinase kinase TAK1 and the MAP kinase NLK counteract Wnt signalling.Results:  To identify NLK‐interacting proteins, we performed yeast two‐hybrid screening. We isolated the gene HMG2L1 and showed that injection of Xenopus HMG2L1 (xHMG2L1) mRNA into Xenopus embryos inhibited Wnt/β‐catenin‐induced axis duplication and expression of Wnt/β‐catenin target genes. Moreover, xHMG2L1 inhibited β‐catenin‐stimulated transcriptional activity in mammalian cells.Conclusions:  Our findings indicate that xHMG2L1 may negatively regulate Wnt/β‐catenin signalling, and that xHMG2L1 may play a role in early Xenopus development together with NLK.
Sphingosine 1‐phosphate promotes cell migration through the activation of Cdc42 in Edg‐6/S1P4‐expressing cells
Authors: Kohno Takayuki , Matsuyuki Hirofumi , Inagaki Yuichi , Igarashi Yasuyuki 
Published: July 2003

AbstractsBackground:  Sphingosine 1‐phosphate (Sph‐1‐P) is a bioactive lipid mediator released from activated platelets, which regulates diverse signal transduction pathways via cell surface receptors. Recent studies have revealed that the seven‐transmembrane‐spanning receptors, Edg‐1, Edg‐3, Edg‐5, Edg‐6 and Edg‐8 are specific Sph‐1‐P receptors. Northern blot analysis has demonstrated that Edg‐6 is expressed in lymphocyte‐containing tissues such as spleen and lung. Little is known about the molecular mechanisms of Edg‐6 functions, probably because of the difficulties in expressing Edg‐6 on the cell surface.Results:  Here, our studies revealed that N‐terminal FLAG‐tagged Edg‐6 or Edg‐6‐GFP fusion protein was expressed in the endoplasmic reticulum, but was not expressed on the cell surface. On the other hand, C‐terminally tagged Edg‐6 or both N‐terminally and C‐terminally tagged Edg‐6 was able to localize to the cell surface. Using these cells, we found that Sph‐1‐P induced cell migration through cell surface‐expressed Edg‐6 in a pertussis toxin‐sensitive manner. This motility was mediated through the activation of a member of the Rho family of small GTPases, Cdc42.Conclusion:  These results support a role for Sph‐1‐P signalling via Edg‐6 in the pathways involved in cell motility.