[Influence of hypergravity on cell proliferation and autogamy immaturity (correction of imaturity) of Paramecium tetraurelia].
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Publications and source records attributed to Y Kato.
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The purpose of this study was to evaluate changes in the hard and soft tissues and to test a preliminary method for predicting the soft tissue profile changes in 14 Japanese retrognathic patients who underwent surgical mandibular advancement. A pair of pre- and postoperative lateral cephalograms was taken for each subject. A paired t test identified significant hard and soft tissue changes following surgery. A Pearson correlation analysis was used to identify soft tissue changes for hard tissue changes in the horizontal and vertical dimensions. Multiple regression models were generated to predict postoperative variables based on preoperative variables. Significant changes in the hard and soft tissues were found in the area inferior to the point Stomion in both the horizontal and vertical dimensions. The soft tissue changes demonstrated a significant correlation not only with the corresponding hard tissue, but also with noncorresponding anatomical points. Moreover, the tendency of changes in the soft tissue was different in the lower lip area from the chin area in both the horizontal and vertical dimensions. Significant R2 and higher contribution ratio were obtained from the multiple regression analysis for the chin area compared to the lower lip area.
The present study determined a suitable parthenogenetic activation procedure for rabbit oocytes and examined the developmental potential of enucleated oocytes receiving cultured cumulus cells. Unfertilized oocytes recovered from superovulated rabbits were activated with one or two sets of electrical pulses, with or without subsequent administration of 6-dimethylaminopurine (6-DMAP). The proportion of oocytes treated with one or two sets of electrical pulses and 6-DMAP that cleaved (87% and 98%, respectively) and developed into blastocysts (77% and 85%, respectively) was significantly higher (P < 0.05) than those activated with electrical pulses alone (30% and 42% for cleavage, 7% and 17% for blastocysts). Cumulus cells separated from ovulated oocytes obtained from mature rabbits were cultured for three to five passages and then induced to quiescence by serum starvation before nuclear transfer. The enucleated oocytes receiving cumulus cells were activated with electrical pulses followed by the addition of 6-DMAP, and cultured in vitro for 5 to 6 d or transferred to pseudopregnant recipient females 1 d after activation. Of 186 nuclear-transferred oocytes, 123 (66%) cleaved and 42 (23%) developed into blastocysts. After transfer of 174 nuclear-transferred oocytes to 8 recipient females, a total of 3 implantation sites were observed in 3 recipient females but no fetuses were obtained.
STUDY DESIGN: A prospective clinical study using venography to evaluate deep venous thrombosis after posterior spinal surgery. OBJECTIVES: To demonstrate the prevalence of deep venous thrombosis after posterior spinal surgery with no prophylaxis. SUMMARY OF BACKGROUND DATA: There have been few studies about the occurrence of deep venous thrombosis after spinal surgery in which venography was used for screening. METHODS: Of the enrolled 134 patients undergoing posterior spinal surgery, 110 (82.1%) were examined with complete surveillance for deep venous thrombosis by venography. There were 64 males and 46 females. The average age at operation was 59.0 years (range, 14-86 years). The levels of the operation were cervical in 54, thoracic in 7, and lumbar in 49. All procedures were performed with patients under general anesthesia. Neither mechanical methods nor anticoagulation medications were used for prophylaxis against thromboembolism. Bilateral ascending venography was performed within 14 days after surgery. RESULTS: There were no patients with clinical signs of deep venous thrombosis and pulmonary embolism. However, 17 patients (15.5%) showed venographic evidence of deep venous thrombosis, of whom 16 had distal thrombi, and only one had a proximal thrombus. Deep venous thrombosis was venographically evident in 3 (5.6%) of 54 patients who underwent cervical procedures, and it was evident in 13 (26.5%) of 49 patients who underwent lumbar procedures. This difference was statistical significant (chi2 test, P = 0.003). Statistical comparison between patients who did and did not have deep venous thrombosis showed that age was statistically significant (Mann-Whitney test; P < 0.05). CONCLUSIONS: The prevalence of deep venous thrombosis after posterior spinal surgery is higher than generally recognized. Therefore, further study is necessary to clarify the appropriate method for screening and the effect of prophylaxis against thromboembolism after spinal surgery.
We show that anthocyanins can function as potent inhibitors of the formation of nitrated tyrosine in vitro, and clarified how pelargonidin (Pel), which has a mono-hydroxyl group on the B-ring, can scavenge peroxynitrite (ONOO(-)) by detection of the reaction products. Pel was reacted with ONOO(-), then the reaction mixture was analyzed using high-performance liquid chromatography (HPLC). The HPLC analyses showed two novel peaks assumed to be the reaction products. Based on the instrumental analyses, the reaction products were identified as p-hydroxybenzoic acid and 4-hydroxy-3-nitrobenzoic acid. Pel can protect tyrosine from undergoing nitration through the formation of p-hydroxybenzoic acid and 4-hydroxy-3-nitrobenzoic acid.
STUDY DESIGN: An in vitro biomechanical study of lumbar spine segments. OBJECTIVE: To study the characteristics of the stiffness test protocol. SUMMARY OF BACKGROUND DATA: In an in vitro study using a flexibility protocol, forces are applied and motions are measured; no center of rotation needs to be specified. In a study using a stiffness protocol, the forces are measured and the motions are applied. This does require the center of rotation to be specified. Many biomechanical studies of the spine are available, but there is lack of clarity concerning which of these two test protocols is appropriate to achieve a certain study goal. METHODS: Five-vertebrae lumbar spine specimens with burst fractures in the middle vertebrae (L1) were used. Specially designed apparatus applied flexion and extension rotations around five centers of rotations located on anteroposterior line through the middle of L1. Maximum moment of 4 Nm was applied. RESULTS: The authors found load-displacement curves, ranges of motion, and neutral zones obtained at the five centers of rotations to be markedly different. The center of rotation located at the posterior longitudinal ligament produced large range of motion and neutral zones in comparison to the centers of rotation located at the anterior longitudinal ligament and the spinous process tip (P<0.01). CONCLUSIONS: The stiffness protocol requires that a center of rotation be specified. Shown here is the significant variability in the load-displacement curves, depending on the choice of the location of the center of rotation. Certain center of rotation locations may block the natural motions of the spine and may result in tissue damage.
We investigated the effects of erythropoietin (EPO) in rat hippocampal and striatal slices, where EPO receptors have been known to exist. EPO stimulated dopamine release from rat striatal slices. Acetylcholine (ACh) release from rat hippocampal slices was not affected by EPO, but high K(+)-induced ACh release was considerably enhanced by EPO. Nitric oxide (NO) production from the hippocampus and the striatum was not affected by EPO. NO-synthase activity was not changed by EPO in the hippocampus or the striatum. These results suggest that EPO stimulates dopamine- and acetylcholine-release without affecting NO production.
The effects of dopamine and L-DOPA on survival were examined in differentiated PC12 cells. Addition of dopamine to the culture medium at 3-30 microM prevented cell death induced by depletion of serum and nerve growth factor (NGF). At 100 microM, dopamine induced cell death. The cell-protective effect of dopamine was not affected by nomifensine, an inhibitor of dopamine uptake, or pargyline, an inhibitor of monoamine oxidase, suggesting that dopamine is working outside the cell. The cell-protective effect of dopamine was blunted by SCH-23390, a D(1) antagonist, but not sulpiride, a D(2) antagonist, indicating that the cell protective effect of dopamine is mediated by D(1) receptors in PC12 cells. L-DOPA also protected PC12 cells from cell death induced by depletion of serum and NGF at low concentrations and showed toxicity at high concentration. The effect of L-DOPA was unchanged after inhibition of conversion of L-DOPA to dopamine by m-hydroxybenzylhydrazine (NSD-1015), an inhibitor of DOPA decarboxylase, suggesting that L-DOPA itself is working for cell protection. Intracellular Ca(2+) concentration and mitogen-activated protein (MAP) kinase activity were increased by both dopamine and L-DOPA. The effects of dopamine and L-DOPA on cell survival were blunted by nicardipine, a Ca(2+) channel blocker, and PD-98059, an inhibitor of MAP kinase kinase (MEK). These results taken together raised the possibility that dopamine and L-DOPA protect PC12 cells from cell death at low concentrations by activating MAP kinase activity via elevation of intracellular Ca(2+) concentration.
The Wnt family of secreted signalling molecules are essential in embryo development and tumour formation. The Frizzled (Fz) family of serpentine receptors function as Wnt receptors, but how Fz proteins transduce signalling is not understood. In Drosophila, arrow phenocopies the wingless (DWnt-1) phenotype, and encodes a transmembrane protein that is homologous to two members of the mammalian low-density lipoprotein receptor (LDLR)-related protein (LRP) family, LRP5 and LRP6 (refs 12-15). Here we report that LRP6 functions as a co-receptor for Wnt signal transduction. In Xenopus embryos, LRP6 activated Wnt-Fz signalling, and induced Wnt responsive genes, dorsal axis duplication and neural crest formation. An LRP6 mutant lacking the carboxyl intracellular domain blocked signalling by Wnt or Wnt-Fz, but not by Dishevelled or beta-catenin, and inhibited neural crest development. The extracellular domain of LRP6 bound Wnt-1 and associated with Fz in a Wnt-dependent manner. Our results indicate that LRP6 may be a component of the Wnt receptor complex.
BACKGROUND: The search for new tumor markers for breast carcinoma has been an area of vigorous study; nonetheless, to the authors' knowledge little new information has emerged beyond the clinical usefulness of CA 15-3. The authors studied serum CYFRA 21-1 in breast carcinoma based on evidence that breast carcinoma expresses cytokeratin 19 fragments and that CYFRA 21-1 is a specific antigen for cytokeratin 19 fragments. METHODS: The serum samples of 86 patients with primary breast carcinoma, 14 patients with recurrent breast carcinoma, 22 patients with benign mammary disease, and 25 healthy controls were provided for measurements of CYFRA 21-1, carcinoembryonic antigen (CEA), and CA 15-3. The relation between clinicopathologic features, prognosis, and disease free survival with serum CYFRA 21-1 titers was studied. RESULTS: There was no difference between the serum CYFRA 21-1 titers from patients with benign mammary disease and those from healthy controls. The sensitivities of CYFRA 21-1 for patients with International Union Against Cancer Stage IV and recurrent tumors were 60% and 64.2%, respectively, which were as high as those for CA 15-3 and superior to those for CEA. The hematogenous recurrence showed a very high sensitivity of 89%. According to the increments of T, N, and M factor numbers, the serum CYFRA 21-1 titers were elevated. No correlation between CYFRA 21-1 and CEA was observed and the correlation between CYFRA 21-1 and CA 15-3 was weak. The univariate and multivariate analyses for survival revealed that serum CYFRA 21-1 levels were an independent indicator of prognosis. CONCLUSIONS: The measurement of the serum CYFRA 21-1 titer in patients with breast carcinoma may be useful in monitoring for recurrence and evaluating the therapeutic effect in patients with advanced disease.
Dityrosine is one of the specific biomarkers for proteinoxidation. We prepared an antibody specific for proteindityrosine using a dimer of 3-p-(hydroxyphenyl)propionic acid (di-HP) as a hapten. Three clones (A8, G6, and 1C3) were obtained, and the antibody from the A8 clone reacted with the di-HP-conjugated protein but not with a dityrosine conjugate. The others (G6 and 1C3 clones) recognized both the di-HP and dityrosine conjugates. The antibodies reacted with peptidyl dityrosine, derived from Thr-Tyr-Ser, rather than the free dityrosine. The reactivity of the latter two antibodies with lens proteins oxidized by incubation with H(2)O(2)/Cu was in good accordance with the formation of the dityrosine-like fluorescence. Using the obtained monoclonal antibody, the immunopositive staining of atherosclerotic lesions in Apo E-deficient mice was confirmed by an immunohistochemical technique.
The crystal structure of glycosyltrehalose trehalohydrolase from the hyperthermophilic archaeum Sulfolobus solfataricus KM1 has been solved by multiple isomorphous replacement. The enzyme is an alpha-amylase (family 13) with unique exo-amylolytic activity for glycosyltrehalosides. It cleaves the alpha-1,4 glycosidic bond adjacent to the trehalose moiety to release trehalose and maltooligo saccharide. Unlike most other family 13 glycosidases, the enzyme does not require Ca(2+) for activity, and it contains an N-terminal extension of approximately 100 amino acid residues that is homologous to N-terminal domains found in many glycosidases that recognize branched oligosaccharides. Crystallography revealed the enzyme to exist as a homodimer covalently linked by an intermolecular disulfide bond at residue C298. The existence of the intermolecular disulfide bond was confirmed by biochemical analysis and mutagenesis. The N-terminal extension forms an independent domain connected to the catalytic domain by an extended linker. The functionally essential Ca(2+) binding site found in the B domain of alpha-amylases and many other family 13 glycosidases was found to be replaced by hydrophobic packing interactions. The enzyme also contains a very unusual excursion in the (beta/alpha)(8) barrel structure of the catalytic domain. This excursion originates from the bottom of the (beta/alpha)(8) barrel between helix 6 and strand 7, but folds upward in a distorted alpha-hairpin structure to form a part of the substrate binding cleft wall that is possibly critical for the enzyme's unique substrate selectivity. Participation of an alpha-beta loop in the formation of the substrate binding cleft is a novel feature that is not observed in other known (beta/alpha)(8) enzymes.
The monoclonal antibody to N(epsilon)-(hexanonyl)lysine (HEL), a novel adduct formed by the reaction of linoleic acid hydroperoxide and lysine, has been prepared and characterized. The obtained antibody specifically recognized the HEL moiety. Using the monoclonal antibody, we evaluated the protective effects of feeding eriocitrin, which is one of flavonoids in lemon fruit, on oxidative modification induced by exercise in rats. The supplementation of eriocitrin significantly suppressed the increase in HEL in the skeletal muscle by exercise. The result suggests that the determination of HEL may be a good method for evaluation of the protective effect of beneficial food factors against oxidative stress.
STUDY DESIGN: The accuracy and safety of atlantoaxial transarticular screw insertion were evaluated in clinical cases. OBJECTIVES: To evaluate the accuracy and safety of atlantoaxial transarticular screw insertion under lateral fluoroscopic monitoring without opening the joint. SUMMARY OF BACKGROUND DATA: Atlantoaxial transarticular screw fixation has been reported to be biomechanically superior to posterior atlantoaxial wiring techniques. Several clinical series have been reported in the literature. In some reports, the risk of screw insertion in this technique has been pointed out. MATERIALS AND METHODS: Fifty-six consecutive patients with atlantoaxial instability were treated by transarticular screw fixation. One hundred twelve screw insertions in these 56 patients were assessed by surgical record and computed tomographic examination. One screw could not be inserted because of the difficulty of adequate placement during operation; 111 screws were therefore inserted. Adequate position was defined as when the screw perforated the lateral atlantoaxial joint. RESULTS: In this series, neither vertebral artery injury nor spinal cord injury was experienced clinically. One guide wire was broken during drilling with a cannulated drill. Computed tomographic examination demonstrated that 106 screws perforated the atlantoaxial joint. Therefore, 95.5% of screws were adequately positioned. There were two screws positioned lateral to the joint, two medially, and one anteroinferiorly to the joint. CONCLUSIONS: Atlantoaxial transarticular screw insertion using image intensifier without opening the lateral joint was performed safely, but not accurately, in all cases.
To investigate the molecular mechanisms of muscle atrophy under microgravity, the paraspinal muscles of rats after 14 days spaceflight and those of ground-based controls were examined. In the microgravitational environment, expressions of 42 genes changed, and the expressions of heat shock protein 70 and t complex polypeptide 1 increased. In Northern blotting, myocyte-specific enhancer binding factor 2C (MEF2C) and MEF2C-related genes including aldolase A and muscle ankyrin decreased. After 9 days ground recovery, expression of MEF2C increased and it was located mainly on the satellite cells in the muscle regeneration state. MEF2C could be a key transcriptional factor for skeletal muscle atrophy and regeneration under microgravity.
Big mitogen-activated protein (MAP) kinase (BMK1), a member of the mammalian MAP kinase family, is activated by growth factors. The activation of BMK1 is required for growth factor-induced cell proliferation and cell cycle progression. We have previously shown that BMK1 regulates c-jun gene expression through direct phosphorylation and activation of transcription factor MEF2C. MEF2C belongs to the myocyte enhancer factor 2 (MEF2) protein family, a four-membered family of transcription factors denoted MEF2A, -2B, -2C, and -2D. Here, we demonstrate that, in addition to MEF2C, BMK1 phosphorylates and activates MEF2A and MEF2D but not MEF2B. The blocking of BMK1 signaling inhibits the epidermal growth factor-dependent activation of these three MEF2 transcription factors. The sites phosphorylated by activated BMK1 were mapped to Ser-355, Thr-312, and Thr-319 of MEF2A and Ser-179 of MEF2D both in vitro and in vivo. Site-directed mutagenesis reveals that the phosphorylation of these sites in MEF2A and MEF2D are necessary for the induction of MEF2A and 2D transactivating activity by either BMK1 or by epidermal growth factor. Taken together, these data demonstrate that, upon growth factor induction, BMK1 directly phosphorylates and activates three members of the MEF2 family of transcription factors thereby inducing MEF2-dependent gene expression.
Colorectal (CRC) and gastric cancers (GC), the most common gastrointestinal malignancies, have been known to develop occasionally in a same patient. Previous studies have focused on the etiology of patients with multiple primary gastric and colorectal cancer (MPGCC); however, the carcinogenic process of MPGCC remains unclear. In this study, we have examined the genetic alterations in MPGCC in order to clarify the carcinogenic pathway. Twenty patients with sporadic MPGCC were examined for microsatellite instability (MSI) and frameshift mutations of target genes such as TGFbetaRII, BAX and IGFIIR. In 10 (50%) of 20 patients with MPGCC, MSI was present at least at 1 lesion of GC or CRC. Four (50%) of 8 cases with synchronous MPGCC displayed MSI in both GC and CRC, while only 1 (8%) of 12 cases of metachronous MPGCC exhibited MSI in both organs. Carcinogenic process of MPGCC was fairly associated with the MSI pathway, particularly in cases of synchronous MPGCC. MSI was found in 5 (25%) of 20 GCs and in 10 (50%) of 20 CRCs. MSI was involved more closely in CRC than in GC among MPGCC. Although most frameshift mutations at target genes were found in the MSI-positive MPGCC, infrequent mutations were observed in the genes. Frameshift mutation was found in only 1 of 5 cases of MSI-positive GC at TGFbetaRII. Only 2 of 10 cases of CRC with MSI showed mutation at TGFbetaRII, and 1 case also showed mutation at BAX and IGFIIR. Our findings suggest that TGFbetaRII, BAX and IGFIIR are not the main target genes for carcinogenesis in MSI-positive MPGCC.
Human Aurora2 was originally identified by its close homology to yeast IPL1 and fly aurora, which are key regulators of chromosome segregation and a family of serine/threonine kinases. Here we demonstrate that the Aurora2 protein is degraded rapidly after G2/M phase release in mammalian cells. Aurora2 protein has a rapid turnover rate with a half-life of approximately 2 h. In eukaryotic cells, the ubiquitin-proteasome pathway is the major mechanism for the targeted degradation of unstable proteins. The treatment of mammalian cells with proteasome inhibitors blocks Aurora2 degradation. Furthermore, Aurora2 is polyubiquitinated in vivo and in vitro using anaphase-promoting complex (APC). These results demonstrate that Aurora2 protein is turned over through the APC-ubiquitin-proteasome pathway. Oncogene (2000) 19, 2812 - 2819