Biomedical subjects

Minoru Fukuda

Publications and source records attributed to Minoru Fukuda.

At least 37 records · Page 2Linked to original sources

PubMed2006

Cellular and molecular analysis of neural development of glycosyltransferase gene knockout mice.

Recent studies demonstrate that carbohydrates synthesized by specific glycosyltransferases play important roles in the development of the central nervous system. Among these carbohydrates, polysialic acid is a unique glycan that modulates functions of the neural cell adhesion molecule (NCAM) by attenuating NCAM-mediated interaction between neural cells. During brain development, polysialic acid is synthesized in a specific spatiotemporal pattern by two polysialyltransferases, ST8SiaII and ST8SiaIV. To study in vivo the roles of polysialic acid synthesized by each respective enzyme, we generated ST8SiaII and ST8SiaIV knockout mice. Single knockout ST8SiaII or ST8SiaIV mice show polysialic acid expression patterns differing from wild type, and those patterns indicate different roles of each gene during neural development. In this chapter, we discuss methods used to analyze polysialyltransferase knockout mice using immunohistochemistry of brain and primary cultures of neurons.

Animals↗

PubMed2006

Mutation of luxS affects motility and infectivity of Helicobacter pylori in gastric mucosa of a Mongolian gerbil model.

Helicobacter pylori is associated with gastric disorders in humans and some experimental animals, and possesses the luxS/type 2 autoinducer (AI-2) system. The effects of a specific luxS mutation on the characteristics of H. pylori were examined. On 0.3 % agar medium, motility of H. pylori HPKY08 (luxS : : cat) was significantly lower than that of wild-type H. pylori TK1402. The luxS-complemented strain HPKY21 exhibited motility comparable to that of H. pylori TK1402. It was shown that the luxS/AI-2 system plays an important role in H. pylori motility. The luxS mutant exhibited a reduced infection rate relative to the wild-type parent strain TK1402 in a Mongolian gerbil model. At 3 months after oral inoculation, lower numbers of H. pylori were detected by semi-quantitative real-time reverse transcription PCR (qRT-PCR) in luxS(-) mutant-infected gerbils than in TK1402-infected gerbils. Gastric inflammation and increased antibody titre for H. pylori were observed in TK1402-infected gerbils only.

Animals↗

PubMed2006

Clinical utility of quantitative RT-PCR targeted to alpha1,4-N-acetylglucosaminyltransferase mRNA for detection of pancreatic cancer.

alpha1,4-N-Acetylglucosaminyltransferase (alpha4GnT) is a glycosyltransferase responsible for the biosynthesis of alpha1,4-GlcNAc-capped O-glycans, and is frequently expressed in pancreatic cancer cells but not peripheral blood cells. In the present study, we tested the clinical utility of alpha4GnT mRNA expressed in the mononuclear cell fraction of peripheral blood as a biomarker of pancreatic cancer. Total RNA isolated from the peripheral blood mononuclear cells from 55 pancreatic cancer patients, 10 chronic pancreatitis patients, and 70 cancer-free volunteers was analyzed quantitatively by reverse transcription-polymerase chain reaction with primers specific for alpha4GnT, and the expression level of alpha4GnT mRNA relative to that of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was measured. When the ratio of alpha4GnT to GAPDH transcripts exceeded a defined cut-off value, patients were considered to have pancreatic cancer. By these standards, 76.4% of the pancreatic cancer patients were detected by this assay. A strong correlation was obtained between positivity in this assay and the expression of alpha4GnT protein detected immunohistochemically in pancreatic cancer tissues resected subsequently, suggesting that alpha4GnT mRNA detected in the peripheral blood is derived from circulating pancreatic cancer cells. Although increased levels of alpha4GnT mRNA was detected in 40.0% of chronic pancreatitis patients and 17.1% of cancer-free volunteers, the expression levels were significantly lower than those seen in pancreatic cancer patients. These results suggest that quantitative analysis of alpha4GnT mRNA expressed in the mononuclear cell fraction of peripheral blood will contribute to the detection of pancreatic cancer.

Adult↗

PubMed2006

Four cases of pulmonary Mycobacterium avium intracellulare complex presenting as a solitary pulmonary nodule and a review of other cases in Japan.

OBJECTIVE: To evaluate clinical findings of patients with a solitary pulmonary nodule in Japan caused by pulmonary Mycobacterium avium complex (MAC) disease. METHODS: The authors investigated the clinical features of 12 patients diagnosed as having pulmonary MAC disease who had presented with a solitary pulmonary nodule. RESULTS: The causative microorganisms were M. avium in seven patients, Mycobacterium intracellulare in two and MAC in three. The diagnostic methods were bronchoscopic biopsy or percutaneous lung biopsy in three patients and surgical operations in the remaining nine. Eleven patients had a complete surgical resection of the nodule and antituberculous drugs were administered to eight. On X-ray, there was an absence of calcification, satellite lesions, cavities, or bronchoectasis that are often thought to be characteristic of pulmonary mycobacterial disease. Differentiation from lung cancer was thought necessary in five patients. There was no microbiological or radiological relapse in those who underwent complete surgical resection. CONCLUSIONS: Because treatment is often poorly effective for patients with pulmonary non-tuberculous mycobacterial disease, it is important to identify the causative microorganisms by performing a culture examination of resected lung tissue especially if there is a solitary pulmonary nodule.

Adult↗

PubMed2006

Chronic necrotizing pulmonary aspergillosis as a complication of pulmonary Mycobacterium avium complex disease.

OBJECTIVE AND BACKGROUND: To investigate the characteristic clinical features of chronic necrotizing pulmonary aspergillosis (CNPA) as a complication of pulmonary Mycobacterium avium complex (MAC) disease. METHODS: Clinical analysis of nine cases without a history of old pulmonary tuberculosis in whom CNPA was found to be a complication during the follow-up period for MAC disease. RESULTS: The average duration from the diagnosis of pulmonary MAC disease to the diagnosis of CNPA was 36.0 months. Five patients received antituberculous therapy including clarithromycin for pulmonary MAC disease, but this treatment was ineffective in most. A positive culture for Aspergillus spp. from sputum and a bronchoscopic specimen and clinical evidence of a chronic infective process were recognized in all cases at the time of detection of CNPA. Serological fungal examinations for anti-Aspergillus IgG antibody were initially negative and became positive in all cases during the follow-up period of pulmonary MAC disease. The presence of CNPA surrounding the cavity previously caused by MAC was characterized by local thickening of the cavity with a fungus ball and the appearance of an infiltration shadow surrounding the cavity. In most of the cases, CNPA was at first treated with oral itriconazole and then with i.v. infusion of micafungin, but the clinical efficacy was generally poor. CONCLUSION: The results of this study showed that during the long follow-up period of patients with pulmonary MAC disease it is important to not only carry out serological examinations, but also perform radiological examinations using chest CT.

Aged↗

PubMed2006

[Clinical usefulness of the (1-->3)-beta-D-glucan measurement kit using the improved alkaline pretreatment method--comparison with conventional method].

We had reported that the rate of non-specific reaction in measurement of (1-->3)-beta-D-glucan was decreased by improvement of the alkaline pretreatment reagent of Fungitec G-test MK (MK assay). To compare the clinical usefulness between conventional MK assay and new MK assay using improved alkaline pretreatment reagent, 121 plasma samples were tested. The sensitivity, specificity, positive predictive value (PPV), negative predictive values (NPV) in conventional MK assay were 91.7%, 85.3%, 44.0% and 98.8%, respectively. Those in new MK assay were 75.0%, 91.6%, 52.9% and 96.7%, respectively. On the other hand, area under the curve (AUC) of receiver operating characteristic (ROC) analysis in conventional and new MK assay was 0.9175 and 0.9123 without a significant difference. It has been recognized that the sensitivity in conventional MK assay is higher than those in other beta-glucan assays. Then, the specificity of new MK assay was improved by using improved alkaline pretreatment reagent, without decreasing the sensitivity. Thus, the present findings indicate that the new MK assay is clinically quite useful.

Adolescent↗

PubMed2006

[Effects of carbon dioxide insufflation during endoscopic conduit harvesting in coronary artery bypass surgery].

BACKGROUND: Endoscopic vessel harvesting is becoming common for the patients undergoing coronary artery bypass grafting. Insufflation of carbon dioxide (CO2) during the procedures has been reported elsewhere to affect arterial carbon dioxide tension, but the occurrence of hypercapnia is still controversial. METHODS: We investigated the effects of CO2 insufflation during endoscopic harvesting of the saphenous vein (SV, n = 34), radial artery (RA, n = 14), or internal mammary artery (IMA, n = 7) for coronary artery bypass surgery. The conduit harvesting was performed using Vasoview Dissecting Cannula with insufflation of CO2 maintaining a cavity pressure of 8-10 mmHg. RESULTS: After insufflation of CO2, significant elevation of partial pressure of CO2 in arterial blood (PaCO2) was found during harvesting of SV (35.4 +/- 3.8 to 49.2 +/- 7.5 mmHg, P < 0.01) and of IMA (38.0 +/- 2.3 to 44.2 +/- 3.2 mmHg, P < 0.05), but no significant elevation of PaCO2 occurred during RA harvesting using a tourniquet. The extent of PaCO2 elevation in SV harvesting showed negative correlation with patient's body weight, body mass index, and body surface area. CONCLUSIONS: Significant hypercarbia occurs during endoscopic harvesting of SV or IMA. It is recommended that PaCO2 should be carefully monitored during endoscopic conduit harvesting for coronary artery bypass surgery.

Aged↗

PubMed2005

Suppression of tumor formation in lymph nodes by L-selectin-mediated natural killer cell recruitment.

Natural killer (NK) cells are known to reject certain tumors in vivo; however, the ability of NK cells to prevent metastasis of tumors into secondary lymphoid organs has not been addressed. Here, we report that in tumor-bearing hosts, NK cells are recruited to regional lymph nodes in wild-type mice, but not in mice deficient for L-selectin or L-selectin ligands. By adoptive transfer and complete Freund's adjuvant stimulation experiments, we demonstrated that L-selectin on NK cells and L-selectin ligands on endothelial cells are essential for NK cell recruitment to lymph nodes. Furthermore, freshly isolated resident lymph node NK cells lysed tumors efficiently, and metastasis of B16 melanoma cells to draining lymph nodes was suppressed in wild-type or Rag-1-deficient mice, but not when NK cells were depleted. Although L-selectin-deficient NK cells efficiently lysed tumor cells in vitro, NK cell-dependent suppression of tumor metastasis was diminished in mice deficient for L-selectin or L-selectin ligands because of insufficient NK cell recruitment to lymph nodes. Moreover, tumor metastasis was substantially inhibited in L-selectin-deficient mice reconstituted with wild-type NK cells. These findings indicate that L-selectin-mediated NK cell recruitment plays a crucial role in the control of tumor metastasis into secondary lymphoid organs.

Adoptive Transfer↗

PubMed2005

N-glycans of core2 beta(1,6)-N-acetylglucosaminyltransferase-I (C2GnT-I) but not those of alpha(1,3)-fucosyltransferase-VII (FucT-VII) are required for the synthesis of functional P-selectin glycoprotein ligand-1 (PSGL-1): effects on P-, L- and E-selectin binding.

C2GnT-I [core2 beta(1,6)-N-acetyglucosaminyltransferase-I] and FucT-VII [alpha(1,3)-fucosyltransferase-VII] are the key enzymes for the biosynthesis of sialyl-Lewis x determinants on selectin ligands and therefore they represent good drug targets for the treatment of inflammatory disorders and other pathologies involving selectins. In the present study, we examined the importance of N-glycosylation for the ability of C2GnT-I and FucT-VII to generate functional selectin ligands, particularly the PSGL-1 (P-selectin glycoprotein ligand-1). We found that (i) both enzymes have their two N-glycosylation sites occupied, (ii) for C2GnT-I, the N-glycan chain linked to Asn-95 significantly contributes to the synthesis of functional PSGL-1 and is required to localize the enzyme to the cis/medial-Golgi compartment, (iii) all N-glycosylation-deficient proteins of FucT-VII displayr a dramatic impairment of their in vitro enzymatic activities, but retain their ability to fucosylate the core2-modified PSGL-I and to generate P- and L-selectin binding, and (iv) the glycomutants of FucT-VII fail to synthesize sialyl-Lewis x or to generate E-selectin binding unless core2-modified PSGL-1 is present. All combined, our results show a differential functional impact of N-glycosylation on C2GnT-1 and FucT-VII and disclose that a strongly reduced FucT-VII activity retains the ability to fucosylate PSGL-1 on the core2-based binding site(s) for the three selectins.

Animals↗

PubMed2005

N-acetylglucosamine-6-O-sulfotransferases 1 and 2 cooperatively control lymphocyte homing through L-selectin ligand biosynthesis in high endothelial venules.

Lymphocyte homing is mediated by specific interactions between L-selectin on lymphocytes and sulfated carbohydrates restricted to high endothelial venules in lymph nodes. Here we generated mice deficient in both N-acetylglucosamine-6-O-sulfotransferase 1 (GlcNAc6ST-1) and GlcNAc6ST-2 and found that mutant mice had approximately 75% less homing of lymphocytes to the peripheral lymph nodes than did wild-type mice. Consequently, these mice had lower contact hypersensitivity responses than those of wild-type mice. Carbohydrate structural analysis showed that 6-sulfo sialyl Lewis X, a dominant ligand for L-selectin, was almost completely absent from the high endothelial venules of these mutant mice, whereas the amount of unsulfated sialyl Lewis X was much greater. These results demonstrate the essential function of GlcNAc6ST-1 and GlcNAc6ST-2 in L-selectin ligand biosynthesis in high endothelial venules and their importance in immune surveillance.

Animals↗

PubMed2005

An A/G polymorphism of core 2 branching enzyme gene is associated with prostate cancer.

The expression of core 2 beta1,6-N-acetylglucosaminyltransferase-1 (C2GnT) is associated with development and progression of malignancy. Sequence analysis showed that the codon 152 of C2GnT has a polymorphism having GTT encoding valine or ATT encoding isoleucine. By examining the polymorphism in prostate cancer and benign prostatic hyperplasia patients, we found that the C2GnT G allele was more frequently observed in the prostate cancer group (p=0.015) than the control group. Men with the GG genotype had a 3.60-fold increased risk of prostate cancer, and men with the AG genotype had a 1.58-fold increased risk of prostate cancer compared with those with the AA genotype. The G allele was found to have a gene dosage effect for prostate cancer risk. No such risk was associated for benign prostatic hyperplasia. These results demonstrate that C2GnT A/G polymorphism is associated with the susceptibility to prostate cancer in a Japanese population.

1,4-alpha-Glucan Branching Enzyme↗

PubMed2005

Expression of core 2 beta1,6-N-acetylglucosaminyltransferase facilitates prostate cancer progression.

Cell surface carbohydrates expressed on epithelial cells are thought to play an important role in tumor progression. Previously, we have shown that expression of core 2-branched O-glycans is closely correlated with vessel invasion and depth of invasion in colon and lung carcinomas. In this study, we found that expression of core 2 beta1,6-N-acetylglucosaminyltransferase-1, Core2GnT, is positively correlated with the progression of prostate cancer in human patients. Statistical analysis demonstrated that Core2GnT is an independent predictor for progressed pathological stage (pT3) and for prostate-specific antigen (PSA) relapse. To determine directly the roles of Core2GnT in prostate cancer progression, we set up an experimental tumor model using the LNCaP prostate cancer cell line. Because this line does not express Core2GnT, we established an LNCaP line stably expressing Core2GnT, LNCap-Core2GnT, by transfecting cDNA encoding Core2GnT. When mock-transfected LNCaP cells and LNCaP-Core2GnT were inoculated in the prostate of nude mice, LNCaP-Core2GnT cells produced three times heavier prostate tumors than mock-transfected LNCaP cells. Furthermore, we found that LNCaP-Core2GnT cells adhered more strongly to prostate stromal cells, type IV collagen and laminin than did LNCaP-mock cells, but LNCaP and LNCaP-Core2GnT cells grew almost at the same rate on plates coated with type IV collagen or laminin. These results indicate that Core2GnT is an extremely useful prognostic marker for prostate cancer progression. The results also suggest that acquiring Core2GnT in prostate carcinoma cells facilitates adhesion to type IV collagen and laminin, and this increased adhesion may be a cause for aggressive tumor formation by prostate cancer cells expressing Core2GnT.

Aged↗

PubMed2005

Polysialic acid facilitates tumor invasion by glioma cells.

Polysialic acid (PSA) is thought to attenuate neural cell adhesion molecule (NCAM) adhesion, thereby facilitating neural cell migration and regeneration. Although the expression of PSA has been shown to correlate with the progression of certain tumors such as small cell lung carcinoma, there have been no studies to determine the roles of PSA in gliomas, the most common type of primary brain tumor in humans. In this study, we first revealed that among patients with glioma, PSA was detected more frequently in diffuse astrocytoma cells, which spread extensively. To determine directly the role of PSA in glioma cell invasion, we transfected C6 glioma cells with polysialyltransferases to express PSA. In those transfected cells, PSA is attached mainly to NCAM-140, whereas the mock-transfected C6 cells express equivalent amounts of PSA-free NCAM-140. Both PSA negative and positive C6 cell lines exhibited almost identical growth rates measured in vitro. However, PSA positive C6 cells exhibited increased invasion to the corpus callosum, where the mock-transfected C6 glioma cells rarely invaded when inoculated into the brain. By contrast, the invasion to the corpus callosum by both the mock-transfected and PSA positive C6 cells was observed in NCAM-deficient mice. These results combined indicate that PSA facilitates tumor invasion of glioma in the brain, and that NCAM-NCAM interaction is likely attenuated in the PSA-mediated tumor invasion.

Astrocytoma↗

PubMed2005

Coexistence of metastatic lung cancer and pulmonary tuberculosis diagnosed in the same cavity.

A 79-year-old woman who had a past history of chronic renal failure 10 years earlier, tongue cancer (T2N2M0) 3 years earlier, and tuberculosis of the cervical lymph nodes 6 months earlier was suddenly admitted with the complaint of right chest pain on April 6, 2004. Right pneumothorax and mild pleural effusion were observed on a chest radiograph. There was no improvement in the patients collapsed lung despite the insertion of a chest drainage tube into the pleural cavity. Three thin-walled cavitary lesions were noted in the right lobe of segment 1 on computed tomography, and the cause of her pneumothorax was thought to be air leakage from the largest cavitary lesion adjacent to the visceral pleura. Partial resection of the right lung by video-assisted thoracoscopic surgery (VATS) was performed at the Department of Thoracic Surgery. Subsequently, it was determined that metastatic squamous cell carcinoma of the lung, corresponding to her tongue cancer, had invaded the visceral pleura adjacent to the largest cavitary lesion. Simultaneously, an epitheloid granuloma with caseating necrosis was observed adjacent to a partially thickened portion of this cavitary lesion. The epitheloid granuloma was found to be acid-fast bacilli-positive and a diagnosis of Mycobacterium tuberculosis pulmonary tuberculosis was made. We report a rare case of the coexistence of metastatic lung cancer originating from tongue cancer and active pulmonary tuberculosis diagnosed in the same large cavitary lesion.

Aged↗

PubMed2005

Community-acquired pneumonia in Japan: a prospective ambulatory and hospitalized patient study.

In this study the aetiology of community-acquired pneumonia (CAP) in Japan was investigated and the incidence of causative pathogens in ambulatory and hospitalized patients was compared. In addition, the roles of Chlamydophila felis and Chlamydophila pecorum as causes of CAP were investigated. Five hundred and six patients with CAP who visited an outpatient clinic or were admitted to one of three different hospitals were enrolled in this study; 106 of them were outpatients and 400 were hospitalized patients. Among the 506 CAP cases, Mycoplasma pneumoniae was the most common pathogen found in the outpatients and Streptococcus pneumoniae was the most common in the hospitalized patients. No cases of Chlamydophila pecorum pneumonia were observed and only one patient had an antibody titre suggestive of recent infection with the feline strain of Chlamydophila. The incidence of infection with M. pneumoniae and Chlamydophila pneumoniae was higher among the outpatients than among hospitalized patients, whereas the incidence of infection with S. pneumoniae and Haemophilus influenzae was higher among the hospitalized patients. Recognition of these results will allow prompt and appropriate antimicrobial therapy to be provided using Japanese CAP guidelines.

Adult↗

PubMed2005

Chlamydia pneumoniae growth inhibition in human monocytic THP-1 cells and human epithelial HEp-2 cells by a novel phenoxazine derivative.

In this study the effects of 2-amino-phenoxazine-3-one (phenoxazine derivate, Phx-3) on Chlamydia (Chlamydophila) pneumoniae growth in human monocytic THP-1 cells as well as human epithelial HEp-2 cells were examined. Cells were infected with bacteria at an m.o.i. of 10 by centrifugation. After washing to remove any remaining bacteria, the cells were incubated with or without Phx-3 in the presence or absence of tryptophan for 72 h. The bacteria in cells were assessed by staining of chlamydial inclusions with FITC-labelled anti-chlamydial antibody, electron microscopic analysis, real-time RT-PCR specific for C. pneumoniae 16S rRNA and propagation on HEp-2 cells. Treatment with Phx-3 significantly inhibited growth of C. pneumoniae in THP-1 and HEp-2 cells. A decrease in the number of bacterial 16S rRNA transcripts was also confirmed in both cell lines by real-time RT-PCR. Electron microscopic studies revealed that treatment with Phx-3 induces bacterial destruction in most of the inclusion bodies in these cells. Addition of tryptophan to the culture slightly blocked the growth inhibition of C. pneumoniae by Phx-3. The reagents did not show any cytotoxicity to the cells at the concentrations used. The results suggest that Phx-3 inhibits C. pneumoniae replication in human monocytic cells as well as epithelial cells, partially depending on the tryptophan-metabolic pathway of host cells. Thus, Phx-3 might be a useful compound for controlling C. pneumoniae growth in cells and may be an alternative conventional therapy.

Aromatase Inhibitors↗

PubMed2005

Outbreak of Chlamydophila pneumoniae infection in long-term care facilities and an affiliated hospital.

This paper reports an outbreak of Chlamydophila pneumoniae infection in long-term care facilities and an affiliated hospital. The outbreak involved rapid spread of infection, and was inconsistent with several outbreaks experienced among younger populations. In addition, there were differences in the incidences among facilities and the affiliated hospital in relation to mean age. Our findings indicate that it is possible that elderly residents may be more susceptible to acquiring this infection. Physicians and other health care providers in long-term care facilities should consider C. pneumoniae in the differential diagnosis of an outbreak of respiratory infection.

Adult↗

PubMed2005

[Non-specific reactions in four methods measuring (1-->3)-beta-D-glucan levels in plasma].

We detected non-specific reactions in the measurement of (1-->3)-beta-D-glucan levels (beta-glucan) in plasma, and the influences of the non-specific reactions on sensitivity and specificity of measurement methods were examined. In this study, 460 plasma samples from 174 patients at Kawasaki Medical School Hospital were used, and the plasma beta-glucan levels were measured by different four methods. The methods included the dilution-heating-endpoint (DHE), dilution-heating-turbidimetric (DHT), alkaline-kinetic (AK), and alkaline-endpoint method (AE) with and without 4-amidinophenyl benzoate hydrochloride (APB) of a protease inhibitor blocking the Limulus reaction. Non-specific reactions were detected from the calculated value under conditions with APB. Therefore, both of the actual values and the values equivalent to non-specific reactions were calculated. The incidence of non-specific reactions was 2.4% in DHE method, 0% in DHT, 53.3% in AK, and 99.3% in AE. The sensitivity and specificity in the methods were 35.7% and 96.0%, 28.6% and 96.0%, 78.6% and 80.1%, and 57.1% and 84.1%, respectively. When subtracted the non-specific reaction values from the actual values in AK and AE method, the specificity was increased by 91.4% and 94.0%, respectively. In these two methods, the non-specific reaction was considered to be a major cause of the low specificity. Finally, to measure plasma beta-glucan levels accurately, non-specific reactions should be excluded as possible by further improvement of measurement methods.

Alkalies↗