Biomedical subjects

Y Takeuchi

Publications and source records attributed to Y Takeuchi.

At least 325 records · Page 18Linked to original sources

PubMed1998

Transport of storage proteins to protein storage vacuoles is mediated by large precursor-accumulating vesicles

Novel vesicles that accumulate large amounts of proprotein precursors of storage proteins were purified from maturing pumpkin seeds. These vesicles were designated precursor-accumulating (PAC) vesicles and had diameters of 200 to 400 nm. They contained an electron-dense core of storage proteins surrounded by an electron-translucent layer, and some vesicles also contained small vesicle-like structures. Immunocytochemical analysis revealed numerous electron-dense aggregates of storage proteins within the endoplasmic reticulum. It is likely that these aggregates develop into the electron-dense cores of the PAC vesicles and then leave the endoplasmic reticulum. Immunocytochemical analysis also showed that complex glycans are associated with the peripheral region of PAC vesicles but not the electron-dense cores, indicating that Golgi-derived glycoproteins are incorporated into the PAC vesicles. These results suggest that the unique PAC vesicles might mediate a transport pathway for insoluble aggregates of storage proteins directly to protein storage vacuoles.

Journal Article↗

PubMed1998

Host range and interference studies of three classes of pig endogenous retrovirus.

Recent interest in the use of porcine organs, tissues, and cells for xenotransplantation to humans has highlighted the need to characterize the properties of pig endogenous retroviruses (PERVs). Analysis of a variety of pig cells allowed us to isolate and identify three classes of infectious type C endogenous retrovirus (PERV-A, PERV-B, and PERV-C) which have distinct env genes but have highly homologous sequences in the rest of the genome. To study the properties of these env genes, expression plasmids for the three env genes were constructed and used to generate retrovirus vectors bearing corresponding Env proteins. Host range analyses by the vector transduction assay showed that PERV-A and PERV-B Envs have wider host ranges, including several human cell lines, compared with PERV-C Env, which infected only two pig cell lines and one human cell line. All PERVs could infect pig cells, indicating that the PERVs have a potential to replicate in pig transplants in immunosuppressed patients. Receptors for PERV-A and PERV-B were present on cells of some other species, including mink, rat, mouse, and dog, suggesting that such species may provide useful model systems to study infection and pathogenicity of PERV. In contrast, no vector transduction was observed on nonhuman primate cell lines, casting doubt on the utility of nonhuman primates as models for PERV zoonosis. Interference studies showed that the three PERV strains use receptors distinct from each other and from a number of other type C mammalian retroviruses.

Animals↗

PubMed1998

A new system for stringent, high-titer vesicular stomatitis virus G protein-pseudotyped retrovirus vector induction by introduction of Cre recombinase into stable prepackaging cell lines.

We report here on stable prepackaging cell lines which can be converted into packaging cell lines for high-titer vesicular stomatitis virus G protein (VSV-G)-pseudotyped retrovirus vectors by the introduction of Cre recombinase-expressing adenovirus. The generated prepackaging cell lines constitutively express the gag-pol genes and contain an inducible transcriptional unit for the VSV-G gene. From this unit, the introduced Cre recombinase excised both a neomycin resistance (Neo(r)) gene and a poly(A) signal flanked by a tandem pair of loxP sequences and induced transcription of the VSV-G gene from the same promoter as had been used for Neo(r) expression. By inserting an mRNA-destabilizing signal into the 3' untranslated region of the Neo(r) gene to reduce the amount of Neo(r) transcript, we were able efficiently to select the clones capable of inducing VSV-G at high levels. Without the introduction of Cre recombinase, these cell lines produce neither VSV-G nor any detectable infectious virus at all, even after the transduction of a murine leukemia virus-based retrovirus vector encoding beta-galactosidase. They reproducibly produced high-titer virus stocks of VSV-G-pseudotyped retrovirus (1.0 x 10(6) infectious units/ml) from 3 days after the introduction of Cre recombinase. We also present evidence that VSV-G-producing cells are still fully susceptible to transduction by VSV-G pseudotypes. However, in this vector-producing system, which regulates VSV-G pseudotype production in an all-or-none manner, the integration of vector DNA into packaging cell lines would be minimized. We further show that heparin significantly inhibits retransduction of VSV-G pseudotypes in the culture fluids of packaging cell lines, leading to a two- to fourfold increase in the yield of the pseudotypes after induction. This vector-producing system was very stable and should be advantageous in human gene therapy.

Animals↗

PubMed1998

Packaging of endogenous retroviral sequences in retroviral vectors produced by murine and human packaging cells.

Interaction of retrovirus vectors and endogenous retroviruses present in packaging cell lines and target cells may result in unwanted events, such as the formation of recombinant viruses and the mobilization of therapeutic vectors. Using sensitive reverse transcriptase PCR assays, we investigated human and murine gene therapy packaging cell lines for incorporation of endogenous retrovirus transcripts into murine leukemia virus (MLV) vector particles and, conversely, whether vector genomes are incorporated into human endogenous retrovirus (HERV) particles. VL30 endogenous retrovirus sequences were efficiently packaged in particles produced by the murine AM12 packaging system. For every seven MLV-derived beta-galactosidase (beta-Gal) vector genomes present in the particles, one copy of VL30 was also packaged. Although human FLY packaging cells expressed several classes of HERV transcripts (HERV-K, HuRT, type C, and RTVL-H), none was detectable in the MLV vector particles released from the cells. Nonspecific packaging of the MLV Gag-Pol expression vector transcripts was detected in the FLY virions at a low level (1 in 17,000 sequences). These findings indicate that human packaging cells produce retrovirus particles far less contaminated by endogenous viral sequences than murine packaging cells. Human teratocarcinoma cells (GH cells), which produce HERV-K particles, were transduced with an MLV-derived beta-Gal vector. Although both HERV-K and RTVL-H sequences were found in association with the particles, beta-Gal transcripts were not detected, indicating that HERV Gag proteins do not efficiently package MLV-based vectors.

Animals↗

PubMed1998

Cationic liposomes enhance the rate of transduction by a recombinant retroviral vector in vitro and in vivo.

Cationic liposomes enhanced the rate of transduction of target cells with retroviral vectors. The greatest effect was seen with the formulation DC-Chol/DOPE, which gave a 20-fold increase in initial transduction rate. This allowed an efficiency of transduction after brief exposure of target cells to virus plus liposome that could be achieved only after extensive exposure to virus alone. Enhancement with DC-Chol/DOPE was optimal when stable virion-liposome complexes were preformed. The transduction rate for complexed virus, as for virus used alone or with the polycation Polybrene, showed first-order dependence on virus concentration. Cationic liposomes, but not Polybrene, were able to mediate envelope-independent transduction, but optimal efficiency required envelope-receptor interaction. When virus complexed with DC-Chol/DOPE was used to transduce human mesothelioma xenografts, transduction was enhanced four- to fivefold compared to that for virus alone. Since the efficacy of gene therapy is dependent on the number of cells modified, which is in turn dependent upon the balance between transduction and biological clearance of the vector, the ability of cationic liposomes to form stable complexes with retroviral vectors and enhance their rate of infection is likely to be important for in vivo application.

3T3 Cells↗

PubMed1998

Epicondylitis among cooks in nursery schools.

OBJECTIVES: To investigate the prevalence and risk factors of epicondylitis among cooks in nursery schools in a cross sectional study because they are suspected to have strenuous workloads on the hands and arms. METHODS: Prevalence of epicondylitis among 209 nursery school cooks and 366 control workers aged 40-59 were studied. Both groups consisted of women workers chosen from 1299 subjects who agreed to participate from 1329 social welfare employees in a city. All workers were interviewed with a questionnaire and had a clinical examination of the tenderness to palpation of epicondyles and epicondylar pain provoked by resisted extension and flexion of the wrist. RESULTS: Nursery school cooks had a significantly higher prevalence of epicondylitis (11.5%) than the controls (2.5%). In a logistic regression model, job title of the cook was also found to have a strong association with epicondylitis (odds ratio (OR) 5.4, 95% confidence interval (95% CI) 2.4 to 11.9) after adjustment for age, body length, and body mass index. Weaker associations were also found between epicondylitis and suspected job stress or workload scores for mechanical workload and psychosocial stressors based on factor analysis. CONCLUSIONS: This study supported the hypothesis that nursery school cooks had a higher prevalence of epicondylitis than other workers with less strenuous hand and arm tasks. It was suggested that risk factors of epicondylitis would be multifactorial, including mechanical workload and psychosocial factors.

Adult↗

PubMed1998

Extrahepatic arterial supply to the liver: observation with a unified CT and angiography system during temporary balloon occlusion of the proper hepatic artery.

PURPOSE: To evaluate routes of potential extrahepatic arterial supply to the liver. MATERIALS AND METHODS: Twenty-three patients with liver tumors underwent computed tomographic (CT) arteriography of extrahepatic arteries before and after temporary balloon occlusion of the proper hepatic artery. The right inferior phrenic artery (RIPA), left inferior phrenic artery (LIPA), superior mesenteric artery (SMA), celiac axis, and left gastric artery (LGA) were evaluated. RESULTS: During temporary balloon occlusion of the proper hepatic artery, extrahepatic arterial supply was immediately evident in 22 of 23 patients (96%). The liver was supplied by the RIPA in 17 of 20 patients (85%), by the LIPA in five of six (83%), by the SMA in eight of 16 (50%), by the celiac axis in two of 10 (20%), and by the LGA in one of six (17%). There was no apparent relationship between the enhanced zones supplied by extrahepatic arteries and the presence or absence of nearby tumors. CONCLUSION: Extrahepatic arterial supply to the liver was readily evident in a large proportion of patients during temporary balloon occlusion of the proper hepatic artery. This finding suggests a need for consideration of extrahepatic arterial supply when angiographic intervention for liver tumors is contemplated.

Aged↗

PubMed1998

Classification based on overnight simultaneous monitoring by electroencephalography and cystometry.

OBJECTIVES: To evaluate the features of the clinical findings in three types (types I, IIa, IIb) of enuresis, classified by overnight simultaneous monitoring by electroencephalography (EEG) and cystometry (CM). METHODS: We performed a test to evaluate bladder capacity, arousal response and the pattern of nighttime urination in sucklings and infants. RESULTS: As for the values of enuretic bladder capacity (EBC), the smallest were found in enuresis type IIb. As for arousal response, enuresis type I showed the best, while enuresis type IIa showed the worst response. With regard to the pattern of nighttime urination in sucklings and infants, all sucklings had a stable bladder in the sleep state and urinated in the pattern of type IIa. The arousal response to bladder distention indicating type I appeared at the age of about 2 or 3. CONCLUSIONS: The smallest EBC in patients with enuresis type IIb represented a bladder dysfunction in the sleep period. It was observed that patients with enuresis type I had the best arousal function while those with enuresis type IIa had the worst arousal function. All sucklings urinated unconsciously during sleep in the pattern of type IIa. The arousal response to bladder distention indicating type I, which was not observed below the age of 1 year, was thought to appear at the age of about 2 or 3 years. If this change has not appeared by the age of 3, bed-wetting might persist after the age of 5.

Child, Preschool↗

PubMed1998

Skin penetration enhancing action of cis-unsaturated fatty acids with omega-9, and omega-12-chain lengths.

The skin penetrative action of high purity cis-omega-12-octadecenoic acid (petroselinic acid, HP-PSA) on rat skin was compared with that of high purity cis-omega-9-octadecenoic acid (oleic acid, HP-OA), following treatment of rat intact skin surface with either 0.05 M HP-PSA or HP-OA in propylene glycol (PG), using Fourier transform/attenuated total reflection (FT-IR/ATR) analysis. Both HP-PSA and HP-OA disordered the lipid structures of the stratum corneum region to a similar extent. Removal of the extractable lipids of the stratum corneum region was marked with HP-PSA/PG but was very slight upon HP-OA/PG treatment. The spectra of the amide II region which originated from proteins suggests that HP-PSA/PG more rapidly disordered the protein structures of both the stratum corneum and the dermis than HP-OA/PG. However, the extent of disordering of the protein structures was presumed to be similar between these two skin penetration enhancers at the maximum level. Enhancement of PG flux in the dermis showed strong positive correlation with the degree of dermis-disordering action of HP-PSA/PG and HP-OA/PG. These results demonstrate that HP-PSA, which has a double bond at an even numbered position (omega-12), more rapidly affects the perturbation of the structures of both the stratum corneum and the dermis than HP-OA, which has the double bond at an odd numbered position (omega-9). Differences in the physicochemical properties of HP-PSA and HP-OA which originate from differences in the double bond position most likely determine the efficacy of these compounds as skin penetration enhancers.

Animals↗

PubMed1998

Focal fatty change in segment IV of the liver occurring after gastrectomy.

In some cases, focal fatty change of the liver parenchyma has been attributed to local systemic venous inflow replacing the portal venous supply. However, no reports of focal fatty change in the dorsal portion of segment IV of the liver caused by direct inflow of an aberrant pancreaticoduodenal vein (APDV) through the parabiliary venous system (PVS) are available. We report a 60-year-old woman with focal fatty change in segment IV of the liver which occurred after gastrectomy with APDV shown on CT performed during selective gastroduodenal arteriography.

Fatty Liver↗

PubMed1998

Effects of certain resorcinol derivatives on the tyrosinase activity and the growth of melanoma cells.

Effects of certain resorcinol derivatives on tyrosinase activity, melanin formation and some other biological activities were studied in order to develop a new, potent depigmentor and/or antimelanoma drug. NKO-09, having isopentyl group in position 6 of resorcinal, exhibited a more potent effect than the compounds which have methyl (NKO-10), ethyl (NKO-11), hexyl (KOM-14), octyl (NKO-14), decyl (NKO-19), dodecyl (NKO-15) and tetradecyl (NKO-16) group in inhibiting the tyrosinase activities (both tyrosine hydroxylation and dopa oxidation). NKO-09 was more potent than hydroquinone in inhibiting the tyrosine hydroxylation; furthermore, NKO-09 inhibited the dopa oxidation different from hydroquinone. In the studies on melanin formation, protein synthesis and the growth of the melanoma cells, NKO-09 caused the most potent effect among the test compounds, except for the growth of the melanoma cells. KOM-14 was more potent than NKO-09 in the antimelanoma activity, and its effect was superior than that of 5-fluorouracil. From these findings, it is suggested that NKO-09 and KOM-14 can be used as an efficacious depigmentor and antimelanoma drug, respectively.

Animals↗

PubMed1998

Reduced expression of interleukin-11 in bone marrow stromal cells of senescence-accelerated mice (SAMP6): relationship to osteopenia with enhanced adipogenesis.

Aging is associated with an increase in bone marrow adipose tissue and a reduction in bone turnover. The P6 strain of senescence-accelerated mice (SAM) exhibit an early decrease in bone mass with a reduction in bone remodeling. In the bone marrow, suppressed osteoblastogenesis and osteoclastogenesis with enhanced adipogenesis are observed. The present study was undertaken to clarify the mechanism of age-related changes in bone turnover using bone marrow cells from SAMP6 mice. Because interleukin (IL)-11 has been shown to potently inhibit adipogenesis and to stimulate osteoclast formation, the effect of IL-11 on the differentiation of bone marrow cells was examined. The impaired formation of both osteoblasts and osteoclasts was restored and the enhanced formation of adipocytes was suppressed by the addition of 10 pM recombinant human IL-11. Other cytokines that activate gp130 as a common signal transducer, IL-6 and leukemia inhibitory factor, did not have such effects. Sequence analysis of the entire coding region of IL-11 cDNA obtained from SAMP6 stromal cells revealed no mutations. Constitutively secreted IL-11 protein into culture media, and its mRNA expression stimulated by transforming growth factor beta were reduced in stromal cells from SAMP6 compared with those in control mice. These results demonstrate that the expression of IL-11 is reduced in bone marrow cells of SAMP6 and suggest that the reduction in IL-11 actions is involved in the impairment of both osteoblastogenesis and osteoclastogenesis in these mice. There is a possibility that alterations in IL-11 actions may be associated with the age-related impairment in bone metabolism.

Adipose Tissue↗

PubMed1998

Morphological variations in transplanted tumors developed by inoculation of spontaneous mesothelioma cell lines derived from F344 rats.

Morphological and immunohistochemical features of the abdominal mesotheliomas that were developed by inoculation of 3 cell lines (MeET-4, -5 and -6) established from spontaneous abdominal mesotheliomas in male F344 rats. Although the original tumors of three cell lines showed signs of epithelioid growth with a predominantly simple papillary pattern, transplanted tumors revealed a variety of morphologic features including epithelioid with glandular structures, sarcomatous, and a mixture of these components. All tumor cells of transplanted tumors were positive for alpha-smooth muscle actin (ASMA) but almost negative for desmin as were epithelioid cells of the original tumors, and the cell lines were positive for desmin but not for ASMA. These results suggested that mesothelioma in the F344 rat had the potential for wide spectrum differentiation under in vitro conditions. The microenvironmental factors obtained in vivo can modify their potential ability and their morphological aspects. These factors may be related to tumor cell reexpression of ASMA of tumor cells that were masked under in vitro culture conditions.

Abdominal Neoplasms↗

PubMed1998

Distribution of F4/80-positive cells in developing ovaries in the mouse.

The mature ovary contains a large number of macrophages. In the present study, the distribution of macrophages in murine ovaries at various developmental stages was immunohistochemically studied using a monoclonal antibody against F4/80, a highly specific antigen of murine macrophages. The results showed that definite F4/80-positive stains were hardly detectable in ovaries on day 0 after birth. On day 7, a few F4/ 80-positive cells could be identified between the developing follicles. The positive stains were irregular in shape and showed little physical contact with the primordial or primary follicles. By days 14 and 21, when the theca cell layers of growing follicles were developing, the positive cells had extended or elongated to surround the cell layer. On day 28, besides the presence of elongating positive cells surrounding the growing follicles, irregularly shaped F4/80-positive cells became apparent in the interstitium between the growing follicles and also in the capsular tissues. Thereafter, positive cells with stellate appearance were detected in the corpora lutea, which first developed around 6 weeks of age. Although the positive cells were homogenously distributed in the corpora lutea in virgin adults, only a few sporadic positive cells were found there in pregnant mice. However, the positive cells infiltrated into the corpora lutea again in the postpartum period. These results show that ovarian macrophages exhibit dramatical changes in their distribution from neonatal to postpartum periods.

Animals↗

PubMed1998

Relationship of occupation to blood pressure among middle-aged Japanese men--the significance of the differences in body mass index and alcohol consumption.

To clarify how and why blood pressure differs between occupations, the proportions of hypertensives, and the measures of blood pressure, body mass index (BMI) and alcohol consumption among the individuals not taking antihypertensive drugs were compared between the eight occupational categories using the data from a health check-up for 589 middle-aged Japanese males, mostly randomly selected from five areas in Japan. After adjusting for age, the relationships of occupation to the proportion of hypertensives and the mean systolic and diastolic blood pressure substantially differed among the five areas. However, after further adjustment for residence, these blood pressure levels (the proportion of hypertensives, and the mean systolic and diastolic blood pressure) were found to be higher for the "Personnel in transport and communications", the "Clerical personnel", the "Managerial and civil personnel" and the "Professional and technical personnel", whereas these values were consistently lowest in the "Service personnel". Age and residence-adjusted mean BMI was also higher for the four occupational categories with the increased blood pressure levels. According to a weighted multiple regression analysis across the eight occupations, the age and residence-adjusted mean BMI was a significant predictor of the age and residence-adjusted mean systolic and diastolic blood pressure (p = 0.068 and 0.018, respectively). These results suggest that the occupation-related changes in BMI may largely contribute to the occupation-related changes in blood pressure.

Alcohol Drinking↗

PubMed1998

Brainstem projections of sensory fibers of the lung: a horseradish peroxidase and c-fos-like immunohistochemical study in the rat.

The brainstem projections of sensory fibers of the lung were determined in the rat by using the horseradish peroxidase (HRP) and c-fos immunohistochemical methods. Wheat germ agglutinin conjugated HRP (WGA-HRP) was injected into the parenchyma of the upper lobe of the left lung. This injection resulted in anterograde labeling in the nucleus of the tractus solitarius (NTS), area postrema (AP) and external cuneate nucleus (ECu) with slightly ipsilateral predominance. It was of interest that these labeled sensory fibers are heavily accumulated in the medial subnucleus at the rostral pole of the NTS and in the commissural subnucleus at the caudal pole. In particular, labeled fibers in the medial subnucleus were characterized by division into the ventral and dorsal portions. After formalin was injected into the parenchyma of the upper lobe of the left lung, the expression of c-fos-like immunoreactivity (FOS-LI) was observed in three nuclei of the brainstem mentioned above. In addition, this experiment resulted in the expression in the ventrolateral medulla, nucleus raphe pallidus and dorsal motor nucleus of vagus nerve bilaterally. With respect to the number of the immunoreactive cells, we could draw the conclusion that the most optimum time to induce the expression of FOS-LI is between 1.5 h and 2.0 h after noxious stimuli.

Animals↗

PubMed1998

Occlusion of the femoral artery induced fos-like immunoreactive neurons in the lateral habenular nucleus projecting to the midbrain periaqueductal gray in the rat.

Wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP) injection into the midbrain periaqueductal gray (PAG) resulted in heavy accumulation of retrogradely labeled neurons in the lateral habenular nucleus (LHb) bilaterally. When the left femoral artery was persistently ligated for 2 h, the expression of Fos-like immunoreactivity (FOS-LI) was also found bilaterally in the LHb. In the present study, by combining the retrograde labeling method of injecting WGA-HRP into the PAG and the immunohistochemical staining of the FOS-LI neurons in the LHb induced by occlusion of the femoral artery, it was demonstrated that there are neurons containing both HRP labeling and FOS-LI. These neurons appeared to be located mainly in the medial part and posterior half of the LHb, and constitute about 24% of the total number of all labeled cells.

Animals↗

PubMed1998

Three-dimensional power Doppler sonography of tumor vascularity.

The purpose of this study was to assess the value of routine clinical examination using three-dimensional power Doppler sonography of intratumoral blood flow. Twenty-two hepatocellular carcinomas, seven cases of hepatic metastasis, four hepatic hemangiomas, six renal cell carcinomas, two cases of hepatic focal nodular hyperplasia, and one case of splenic metastasis were included in the study. Three-dimensional images were reconstructed by maximum intensity projection method using cine-loop data on a built-in computer in a LOGIQ 500 and a LOGIQ 700 from GE Yokogawa Medical Systems. The three-dimensional images obtained were viewed multidirectionally on a monitor screen. Three-dimensional representations of intratumoral blood flow became available for all tumors approximately 5 s to 30 s after scanning. In every case, the entire vasculature of the tumor was appreciated more easily from three-dimensional images than from cross-sectional two-dimensional images. These three-dimensional images of intratumoral blood flows corresponded to the tumor vessels that could be visualized by angiography at the early arterial phase. Differential diagnosis of hepatic tumors based on distinct difference in their intratumoral vascular structures was performed. Our results suggest that three-dimensional power Doppler sonography can be used for routine clinical examination of tumor vascularity and may provide improved diagnostic information.

Carcinoma, Hepatocellular↗