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Biomedical subjects

K Kitajima

Publications and source records attributed to K Kitajima.

At least 37 records · Page 2Linked to original sources
PubMed2001

In vivo function of a differentiation inhibitor, Id2.

Cell differentiation is an essential process for the development of various cell types that constitute multicellular organisms. During development, the large family of factors bearing a helix-loop-helix (HLH) motif participates profoundly in this process and these factors serve as good experimental tools for investigating mechanisms underlying tissue-specific differentiation. The HLH family includes both positive and negative regulators of cell differentiation: basic HLH (bHLH)-type transcription factors and Id proteins, respectively. Following an exciting decade focusing on bHLH factors, advances achieved in studies of the inhibitory factors in the last couple of years have placed them in the front line of the research on differentiation and proliferation control. Here, we present and discuss recent results obtained using Id2-deficient mice, which manifest intriguing phenotypes in various systems.

Animals↗
PubMed2001

Molecular cloning of a unique CMP-sialic acid synthetase that effectively utilizes both deaminoneuraminic acid (KDN) and N-acetylneuraminic acid (Neu5Ac) as substrates.

2-keto-3-deoxy-D-glycero-D-galacto-nononic acid (KDN) is a sialic acid (Sia) that is ubiquitously expressed in vertebrates during normal development and tumorigenesis. Its expression is thought to be regulated by multiple biosynthetic steps catalyzed by several enzymes, including CMP-Sia synthetase. Using crude enzyme preparations, it was shown that mammalian CMP-Sia synthetases had very low activity to synthesize CMP-KDN from KDN and CTP, and the corresponding enzyme from rainbow trout testis had high activity to synthesize both CMP-KDN and CMP-N-acetylneuraminic acid (Neu5Ac) (Terada et al. [1993] J. Biol. Chem., 268, 2640-2648). To demonstrate if the unique substrate specificity found in the crude trout enzyme is conveyed by a single enzyme, cDNA cloning of trout CMP-Sia synthetase was carried out by PCR-based strategy. The trout enzyme was shown to consist of 432 amino acids with two potential nuclear localization signals, and the cDNA sequence displayed 53.8% identity to that of the murine enzyme. Based on the Vmax/Km values, the recombinant trout enzyme had high activity toward both KDN and Neu5Ac (1.1 versus 0.68 min(-1)). In contrast, the recombinant murine enzyme had 15 times lower activity toward KDN than Neu5Ac (0.23 versus 3.5 min(-1)). Northern blot analysis suggested that several sizes of the mRNA are expressed in testis, ovary, and liver in a tissue-specific manner. These results indicate that at least one cloned enzyme has the ability to utilize both KDN and Neu5Ac as substrates efficiently and is useful for the production of CMP-KDN.

Amino Acid Sequence↗
PubMed2001

Morphological and biochemical changes of isolated chicken egg-envelope during sperm penetration: degradation of the 97-kilodalton glycoprotein is involved in sperm-driven hole formation on the egg-envelope.

The chicken egg-envelope is made of two major glycoprotein components, which are designated as gp97 and gp42 (after their molecular masses). To elucidate how these two components are involved in macromolecular organization of the chicken egg-envelope, the isolated egg-envelope was characterized by immunochemical and biochemical methods. The gp97 was suggested to be a homologue of mouse ZPB based on the similarities of N-terminal and internal sequences. Immunoblotting using anti-gp97 monoclonal antibodies and two-dimensional gel electrophoresis with or without mercaptoethanol treatment revealed that gp97 formed a homodimer through disulfide bonds, whereas gp42 did not. Under indirect immunofluorescence microscopy, the anti-gp97 antibody visualized indistinct, small spots on the egg-envelope, whereas the anti-gp42 antibody showed a meshwork of blurry, fibrous structures. The hole formation on the egg-envelope by in vitro sperm penetration was completely inhibited by two anti-gp97 monoclonal antibodies. Interestingly, the anti-gp97 monoclonal antibodies blocked the proteolysis not only of gp97 but also of gp42 during incubation of the egg-envelope with either sperm or the crude chicken acrosin. Taken together, these results indicate that gp97 may play pivotal roles not only in constitution of the macromolecular organization of the egg-envelope but also in triggering hydrolysis of the egg-envelope during sperm penetration.

Amino Acid Sequence↗
PubMed2001

Effects of transglottal pressure on fundamental frequency of phonation: study with a rubber model.

The fundamental frequency of phonation (F0) change per unit change in transglottal pressure (dF/dP) was studied at different extension lengths and masses of the vibrating part of a rubber model. The dF/dP was decreased when the vibrating part was extended, and also when the mass of the vibrating part was increased. In the former, the F0 was increased, and in the latter, the F0 was decreased. In humans, we have noticed that when the F0 was increased, the dF/dP first decreased and then increased, taking a V-shaped curve as a function of the F0. The present study reveals the mechanism underlying the V-shaped curve observed in humans.

Glottis↗
PubMed2001

Effects of length and depth of vibration of the vocal folds on the relationship between transglottal pressure and fundamental frequency of phonation in canine larynges.

The relationship between fundamental frequency of phonation (F0) and the change in F0 per unit change of transglottal pressure (dF/dP) was investigated in 3 excised canine larynges. The effects of the length and depth of vocal fold vibration on the relationship between dF/dP and F0 were evaluated. A positive correlation between dF/dP and F0 was observed when an increase in F0 was accompanied by a decrease in the depth of vibration while the length of vibration reached a plateau. Although a negative correlation between dF/dP and F0 was observed in only 1 case, this relationship seemed relevant to the length of vibration. These results help to clarify the adjustments of length and depth of vibration involved in regulating F0.

Animals↗
PubMed2001

Relationship between transglottal pressure and fundamental frequency of phonation, with effects of dehydration produced by atropine, in healthy volunteers.

We investigated the effects of the intravenous injection of atropine sulfate on the relationship between the fundamental frequency of phonation (F0) and the magnitude of F0 change per unit change in transglottal pressure (dF/dP) in 4 healthy male volunteers. Before the injection of atropine, we observed a nonlinear or V-shaped relationship between dF/dP and F0. After the injection, the values for dF/dP were reduced at a lower F0, whereas the values were unchanged at a higher F0. The results support our hypothesis that the relationship between dF/dP and F0 reflects adjustments in the length and depth of the vocal folds in controlling F0.

Adult↗
PubMed2001

Deficiency of phospholipase C-gamma1 impairs renal development and hematopoiesis.

Phospholipase C-gamma1 (PLC-gamma1) is involved in a variety of intracellular signaling via many growth factor receptors and T-cell receptor. To explore the role of PLC-gamma1 in vivo, we generated the PLC-gamma1-deficient (plc-gamma1(-/-)) mice, which died of growth retardation at embryonic day 8.5-9.5 in utero. Therefore, we examined plc-gamma1(-/-) chimeric mice generated with plc-gamma1(-/-) embryonic stem (ES) cells for further study. Pathologically, plc-gamma1(-/-) chimeras showed multicystic kidney due to severe renal dysplasia and renal tube dilation. Flow cytometric analysis and glucose phosphate isomerase assay revealed very few hematopoietic cells derived from the plc-gamma1(-/-) ES cells in the mutant chimeras. However, differentiation of plc-gamma1(-/-) ES cells into erythrocytes and monocytes/macrophages in vitro was observed to a lesser extent compared with control wild-type ES cells. These data suggest that PLC-gamma1 plays an essential role in the renal development and hematopoiesis in vivo.

Animals↗
PubMed2000

Endoscopic surgery for lateral cervical cysts. A report of three cases.

Recently, endoscopic surgery has been applied to cervical exploration. We have developed new techniques for endoscopic neck surgery, and in this paper report our experience with three patients with lateral cervical cysts. A 5- or 10-mm midline trocar for the endoscope and two 5- or 10-mm lateral trocars were inserted from the anterior chest wall and/or both axillary fossae to avoid neck scars. There were no intraoperative complications. Slight subcutaneus emphysema was present postoperatively, but it was limited to the neck and disappeared in a few days. The incisions were completely covered by the patients' undergarments. This is the first report of endoscopic lateral cervical cystectomy.

Adolescent↗
PubMed2000

Molecular cloning and expression of the mouse N-acetylneuraminic acid 9-phosphate synthase which does not have deaminoneuraminic acid (KDN) 9-phosphate synthase activity.

A cDNA of the mouse homologue of Escherichia coli N-acetylneuraminic acid (Neu5Ac) synthase (neuB gene product) was cloned by the PCR-based method. The mouse homologue consists of 359 amino acids, and the cDNA sequence displays 33% identity to that of the E. coli Neu5Ac synthase. The recombinant mouse homologue which is transiently expressed in HeLa cells does not exhibit the Neu5Ac synthase activity, which catalyzes condensation of phosphoenolpyruvate (PEP) and N-acetylmannosamine (ManNAc) to synthesize Neu5Ac, but the Neu5Ac 9-phosphate (Neu5Ac-9-P) synthase activity, which catalyzes condensation of PEP and ManNAc 6-phosphate (ManNAc-6-P) to synthesize Neu5Ac-9-P. Thus, the mouse homologue of E. coli Neu5Ac synthase is the Neu5Ac-9-P synthase. The Neu5Ac-9-P synthase is a cytosolic enzyme and ubiquitously distributed in mouse various tissues. Notably, the Neu5Ac-9-P synthase can not catalyze the synthesis of deaminoneuraminic acid (KDN) or KDN-9-P from PEP and Man or ManNAc-6-P, thus suggesting that the enzyme is not involved in the synthesis of KDN. This is consistent with the previous observation that only a very low activity to synthesize KDN is found in mouse B16 cells [Angata, T., et al. (1999) Biochem. Biophys. Res. Commun. 261, 326-331].

Amino Acid Sequence↗
PubMed2000

Frequent occurrence of pre-existing alpha 2-->8-linked disialic and oligosialic acids with chain lengths up to 7 Sia residues in mammalian brain glycoproteins. Prevalence revealed by highly sensitive chemical methods and anti-di-, oligo-, and poly-Sia antibodies specific for defined chain lengths.

The pre-existence of alpha2-->8-linked disialic acid (di-Sia) and oligosialic acid (oligo-Sia) structures with up to 7 Sia residues was shown to occur on a large number of brain glycoproteins, including neural cell adhesion molecules (N-CAMs), by two highly sensitive chemical methods (Sato, C., Inoue, S., Matsuda, T., and Kitajima, K. (1998) Anal. Biochem. 261, 191-197; Sato, C., Inoue, S., Matsuda, T., and Kitajima, K. (1999) Anal. Biochem. 266, 102-109). This unexpected finding was also confirmed using a newly developed antibody prepared using a copolymer of alpha2-->8-linked N-acetylneuraminyl p-vinylbenzylamide and acrylamide as an immunogen and known antibodies whose immunospecificities were determined to be di- and oligo-Sia residues with defined chain lengths. The major significance of the new finding that di- and oligo-Sia chains exist on a large number of brain glycoproteins is 2-fold. First, it reveals a surprising diversity in the number and M(r) of proteins distinct from N-CAM that are covalently modified by these short sialyl glycotopes. Second, it suggests that synthesis of di- and/or oligo-Sia units may be catalyzed by alpha2-->8-sialyltransferase(s) that are distinct from the known polysialyltransferases, STX and PST, which are partially responsible for polysialylation of N-CAM.

Animals↗
PubMed2000

Spontaneous galactosylation of agalactoglycoproteins in colostrum.

We have found that spontaneous galactosylation of GlcNAc residues occurs in bovine colostrum, but not in dialyzed colostrum, without adding UDP-Gal as a donor substrate. UDP-Gal was shown to be present in bovine colostrum at a level ranging from 200 to 600 microM. When a tracer UDP-[(14)C]Gal was added to the dialyzed colostrum together with a Gal beta1,4-specific beta-galactosidase, remarkable incorporation of radioactivity into 24-28 kDa and 33 kDa RCA1-positive glycoproteins was demonstrated by SDS-PAGE/autoradiography. Some 100-140 kDa agalactoglycoproteins of a CHO mutant cell line were also galactosylated on a blotted membrane by the incubation in the colostrum.

Animals↗
PubMed2000

Factors affecting survival after living-related liver transplantation.

The purpose of this study was to determine the perioperative factors that influence patient and graft outcome in living-related liver transplantation (LRLT). Between April 1995 and October 1998, we performed a series of 46 LRLT procedures, including 11 adult cased, at our institute. Mean age and weight of the recipients were 12.0 +/- 2.3 years and 23.7 +/- 2.6 kg, respectively. Seven out of the 46 patients had renal failure and received hemodialysis therapy before and after LRLT or kidney transplantation. The recipients were divided into two groups: those who survived for 7-48 months after LRLT (group 1, n = 36), and those who died within 4 months after surgery (group 2, n = 10). Factors analyzed included recipient age and weight, graft/recipient body weight ratio (G/R ratio), emergent vs elective surgery, United Network for Organ Sharing (UNOS) status, presence of preoperative plasmapheresis (PEX) and renal failure, and so on. Recipients in group 1 compared with group 2 had less advanced liver disease (i. e., a lower rate of emergent surgery, 14% vs 50%, and fewer patients with UNOS status 1, 14% versus 70%; P < 0.05 and P < 0.001, respectively). Group 1 recipients also had a lower percentage of preoperative treatment with plasmapheresis (22% vs 70%, P < 0.01). However, neither the G/R ratio nor the presence of renal failure affected the patient survival rate. In conclusion, factors independently associated with reduced patient survival after LRLT include emergent surgery, Child-Pugh class, UNOS status 1, and preoperative plasmapheresis.

Adolescent↗
PubMed2000

Effects of endolymphatic-perilymphatic fistula on endolymphatic hydrops in guinea pig.

The endolymphatic-perilymphatic shunt operation between the scala media and scala tympani was performed in 22 guinea pigs with endolymphatic hydrops induced by the silver nitrate injection method 2 months before the operation. Two (n = 10), 7 (n = 6), and 28 (n = 6) days after the operation, the condition of the fistula and the inner ear pathology were studied histopathologically. In half of the animals, the fistula was open, whereas in the other half, the fistula was closed. In the group examined after 2 postoperative days, the animals with open fistulas showed a slight collapse of Reissner's membrane, which might indicate that excess endolymph escaped through the fistula into the scala tympani. In the groups examined later, especially in the group examined after 28 postoperative days, the animals showed distention of hydrops despite preservation of the fistula. Moderate-to-severe degenerative changes of the organ of Corti, macula sacculi, and stria vascularis were observed in the vicinity of the fistula. We conclude from this study that the endolymphatic-perilymphatic fistula created inside the cochlea apparently was not sufficient to reduce the hydrops. Rather, it caused degenerative changes of the inner ear structure. This type of surgery is not suitable for Meniere's disease.

Animals↗
PubMed2000

C-type natriuretic peptide-like immunoreactivity in the rat inner ear.

C-type natriuretic peptide (CNP) is a member of the atrial natriuretic peptide family (ANP family). The family also includes ANP and brain natriuretic peptide (BNP). These peptides regulate the homeostasis of body fluid and blood pressure as a neuropeptide in the central nervous system as well as a cardiac hormone in the periphery. We have recently reported the expression of CNP mRNA in the inner ear. To assess the possible physiological role of CNP in the inner ear, we investigated the localization of CNP peptide in the rat inner ear by immunohistochemistry at the light and electron microscopic level. CNP-like immunoreactivity was widely distributed in the secretory and the neuronal portion of the inner ear, i.e. the spiral ligament, the dark cell region of the utriculus, the epithelium of the endolymphatic sac, the spiral ganglion cells and the vestibular ganglion cells. The results suggest that CNP may play a role in the homeostasis of the perilymph and endolymph and may also influence nerve activities in the inner ear.

Animals↗
PubMed2000

Laryngeal closure pressure during phonation in humans.

We investigated the laryngeal closure pressure during sustained phonation in five healthy adult men with no pathological lesions in the glottis using a round pressure transducer with a diameter of 10 mm and a thickness of 1 mm. The transducer was placed between the vocal processes through the mouth. Subjects were asked to utter a sustained phonation of the vowel sound /eh/ at varying pitch and intensity in modal register. The laryngeal closure pressure ranged from 0 to 200 cmH2O. At a comfortable pitch and intensity, the pressure was below 50 cmH2O. The pressure was correlated with pitch, but not with intensity.

Adult↗