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Biomedical subjects

H Katayama

Publications and source records attributed to H Katayama.

At least 19 recordsLinked to original sources

Induction of apoptosis in an estrogen-responsive mouse Leydig tumor cell by leukotriene.

For estrogen-responsive B-1F cells, established from estrogen-responsive mouse Leydig cell tumor, it has been reported that the 5-lipoxygenase (5-LOX) metabolic pathway appears to be associated with cell growth. The addition of 5-LOX inhibitor 2-(12-hydroxydodeca-5,10-diyl)-3,5,6-trimethyl-1,4-benzoquinone (AA861) to the medium resulted in a dose-dependent increase in cell yield as described previously. When the growth of the palpable tumors was measured, AA861 had stimulated in vivo tumor growth in adult male mouse inoculated B-1F cells. The effects of AA861 and 17beta-estradiol (E2) on the contents of various arachidonic acid metabolites in B-1F cells and their conditioned medium were examined. Although AA861 and E2 decreased the contents of leukotrienes (LTs), the two did not significantly change those of prostaglandins, thromboxan, prostacyclin, 12-hydroxyeicosatetraenoic acid (HETE) and 15-HETE. In immunohistochemical study B-1F cells show positive staining for 5-LOX in the E2-depleted condition, while E2 decreased the expression of 5-LOX. The decrease of the intensities of 79-kDa 5-LOX protein and 403-bp RT-PCR product bands was observed. The growth of Morpholino-anti oligo delivered B-1F cells was higher than that of Standard control oligo delivered cells. The delivery of Morpholino-anti oligo into B-1F cells caused the decrease of contents of LTs and 5-HETE in the cells and medium, and the reduction of 5-LOX activity. When LTD4 was added in the culture medium, the increasing concentrations of LTD4 resulted in a significant inhibition of cell yields of E2-treated B-1F cells. Morphological changes such as nuclear condensation and fragmentation, and DNA ladder pattern were demonstrated in E2-stimulated B-1F cells treated with LTD4 as well as in control cells cultured in the basal medium. These results implicate that 5-LOX at least plays an important role in the growth of B-1F cells and LD4 induces the apoptosis of B-1F cells.

Animals↗

Calcium carbonate deposition in a cell wall sac formed in mulberry idioblasts.

Although calcium carbonate is known to be a common biomineral in plants, very little attention has been given to the biological control of calcium carbonate deposition. In mulberry leaves, a subcellular structure is involved in mineral deposition and is described here by a variety of cytological techniques. Calcium carbonate was deposited in large, rounded idioblast cells located in the upper epidermal layer of mulberry leaves. Next to the outmost region ("cap") of young idioblasts, we found that the inner cell wall layer expanded to form a peculiar outgrowth, named cell wall sac in this report. This sac grew and eventually occupied the entire apoplastic space of the idioblast. Inside the mature cell wall sac, various cellulosic membranes developed and became the major site of Ca carbonate deposition. Concentrated Ca2+ was pooled in the peripheral zone, where small Ca carbonate globules were present in large numbers. Large globules were tightly packed among multiple membranes in the central zone, especially in compartments formed by cellulosic membranes and in their neighboring membranes. The maximum Ca sink capacity of a single cell wall sac was quantified using enzymatically isolated idioblasts as approximately 48 ng. The newly formed outgrowth in idioblasts is not a pure calcareous body but a complex cell wall structure filled with substantial amounts of Ca carbonate crystals.

Calcium↗

Canine distemper virus induces apoptosis through caspase-3 and -8 activation in vero cells.

We investigated the signal-transduction pathway of canine distemper virus-Onderstepoort (CDV-Ond) vaccine strain-mediated apoptosis in Vero cells. Canine distemper virus-Onderstepoort at a multiplicity of infection (MOI) of 0.1 induced DNA fragmentation 48 h after infection. Immunofluorescence staining revealed that 57% +/- 4% of the CDV-N-protein-positive cells were terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive, and all TUNEL-positive cells were CDV-N-protein-positive, indicating that CDV-Ond induced apoptosis only in the infected cells. We also found that CDV-Ond infection induced activation of caspase-3 and caspase-8. In the semi-quantitative reverse transcription-polymerase chain reaction assay for apoptosis-related genes, the expression of mRNA of the death receptor, Fas, was also increased in CDV-Ond-infected cells. In contrast, the expressions of Bcl-2 and Bax, regulators for intrinsic apoptotic signaling through the mitochondria, did not change. These results suggest that CDV-Ond induced apoptosis by activating caspase-3, possibly through caspase-8 signaling rather than through p53/Bax-mediated, mitochondrial signaling in the infected cells.

Animals↗

Accumulation of copper and silver onto cell body and its effect on the inactivation of Pseudomonas aeruginosa.

Pseudomonas aeruginosa, a gram-negative rod bacterium, is a causative agent of waterborne pneumonia and presents high tolerance against conventional disinfectants. The inorganic biocidal reagents, copper and silver, were applied to inactivate P. aeruginosa inoculated in a synthetic drinking water (SDW). Additionally, the relationship of the specific amount of accumulated copper and silver reagents (Cs) on P. aeruginosa with inactivation profile was elucidated in this study. Flow cytometry (FCM) following staining with SYTO 9 and PI was used for detection of bacterial viability and density. Individual copper and silver reagents, and their combination, exhibited excellent biocidal abilities even at the concentration of 0.05 mgCu/L and 0.005 mgAg/L. The critical amounts of accumulated disinfectant (Cs) were calculated at 2.82 x 10(-7) microgCu/cells and 5.13 x 10(-8) microgAg/cells; at an incubation of 70 h. Consequently, the role of disinfectant on the inactivation of P. aeruginosa and the assessment of biocidal ability of copper, silver, and their combination were successfully explained by evaluating the terms Cs and Cc.

Copper↗

Quantification and genotyping of Cryptosporidium spp. in river water by quenching probe PCR and denaturing gradient gel electrophoresis.

A new detection method was developed for the simultaneous quantification and genotyping of Cryptosporidium spp. in river water. Several modifications made to the US EPA Method 1623 enabled high and stable recovery of Cryptosporidium from 40 L of river water (geometric mean = 35%, standard deviation = 8.7%). Quenching probe PCR (QProbe PCR) was used to quantify the 18S rRNA gene of Cryptosporidium spp. This method could successfully detect single oocysts in a sample, and the lower quantitation limit was as low as 2.5 oocysts/sample. In addition, denaturing gradient gel electrophoresis (DGGE) followed by DNA sequencing was used to identify the genotypes. These methods were applied to detect Cryptosporidium spp. in the Koyama River, Japan. The positive ratio was 69% (11/16) with the maximum concentration of 59 oocysts/100 L. Seven genotypes including two novel ones were identified. These results showed that this detection method could provide valuable information on Cryptosporidium in river water, both in the concentration and in the genotypes, which is essential for the precise assessment of waterborne risk to human health.

Animals↗

Monitoring of human enteric viruses and coliform bacteria in waters after urban flood in Jakarta, Indonesia.

Floodwaters in Kampung Melayu village, Jakarta, Indonesia, as well as river water and consumable water (including groundwater and tap water) samples in flooded and non-flooded areas, were quantitatively analysed to assess occurrence of viruses and total coliforms and E. coli as bacterial indicators after flooding event. High numbers of enterovirus, hepatitis A virus, norovirus (G1, G2) and adenovirus were detected at high concentration in floodwaters and waters sampled from Ciliwung River which runs across metropolitan Jakarta and is used widely for agriculture and domestic purposes by poor residents. One out of three groundwater wells in the flooded area was contaminated with all viruses tested while no viruses were found in groundwater samples in non-flooded areas and tap water samples. The results revealed that human enteric viruses, especially hepatitis A virus and adenovirus, were prevalent in Jakarta, Indonesia. This study suggested that flooding posed a higher risk of viral infection to the people through contamination of drinking water sources or direct contact with floodwaters.

Base Sequence↗

Effects of rainfall on the occurrence of human adenoviruses, total coliforms, and Escherichia coli in seawater.

A two-month survey was conducted in order to evaluate the effects of rainfall on the fate of microorganisms in seawater in the Tokyo Bay, Japan. The seawater sample (1,000 mL) was applied to a method to concentrate virus, followed by a quantification of human adenoviruses using the real-time PCR. Total coliforms and E. coli, which were determined by the colony forming method, were detected in all 47 seawater samples, while human adenoviruses were detected in 38 (81%) of the samples. The concentration of tested microorganisms showed 1-2 log units increase after rainfall events, followed by the gradual decrease to the level before the rainfall within a few days.

Adenoviridae↗

Structural analysis of chloroplast DNA in Prunus (Rosaceae): evolution, genetic diversity and unequal mutations.

In order to understand the evolutionary aspects of the chloroplast DNA (cpDNA) structures in Rosaceous plants, a physical map of peach (Prunus persica cv. Hakuhou) cpDNA was constructed. Fourteen lambda phage clones which covered the entire sequence of the peach cpDNA were digested by restriction enzymes (SalI, XhoI, BamHI, SacI, and PstI) used singly or in combination. The molecular size of peach cpDNA was estimated to be about 152 kb. The gene order and contents were revealed to be equivalent to those of standard type of angiosperms by the localization of 31 genes on the physical map. Eighteen accessions from 14 Prunus species (P. persica, P. mira, P. davidiana, P. cerasis, P. cerasifera, P. domestica, P. insititia, P. spinosa, P. salicina, P. maritima, P. armeniaca, P. mume, P. tomentosa, P. zippeliana, and P. salicifolia) and one interspecific hybrid were used for the structural analysis of cpDNAs. Seventeen mutations (16 recognition site changes and one length mutation) were found in the cpDNA of these 18 accessions by RFLP analysis allowing a classification into 11 genome types. Although the base substitution rate in the recognition site (100p = 0.72) of cpDNA in Prunus was similar to that of other plants, i.e., Triticum-Aegilops, Brassica, and Pisum, it differed from Pyrus (100p = 0.15) in Rosaceae. Seven mutations including one length mutation were densely located within a region of about 9.1 kb which includes psbA and atpA in the left border of a large single-copy region of Prunus cpDNAs. The length mutation was detected only in P. persica and consisted of a 277 bp deletion which occurred in a spacer region between the trnS and trnG genes within the 9.1 kb region. Additional fragment length mutations (insertion/deletion), which were not detected by RFLP analysis, were revealed by PCR and sequence analyses in P. zippeliana and P. salicifolia. All of these length mutations occurred within the 9.1 kb region between psbA and atpA. This region could be an intra-molecular recombinational hotspot in Prunus species.

Base Sequence↗

Adoptive transfer of T-helper cell type 1 clones attenuates an asthmatic phenotype in mice.

T-helper cell type 1 (Th1) cells have been postulated to have a significant role in protective immunity against allergic diseases. However, recent studies using polarised Th1 cells showed conflicting effects on both airway responsiveness and eosinophilic inflammation in a mouse asthma model. The current study explored the effects of adoptive transfer of established Th1 clones on a murine model of atopic asthma. Mice (BALB/c) were sensitised with ovalbumin (OVA) and challenged with aerosolised OVA (5%, 20 min) for 5 days. Just before starting the first challenge, Th1 clones (5x10(6) x body(-1)) or PBS alone were injected via the tail vein. After assessment of airway responsiveness to methacholine, bronchoalveolar lavage fluid (BALF) was obtained. Histological examination, including morphometric analysis, measurement of cytokines in the BALF and Northern blotting of lung chemokines, was also performed. Adoptive transfer of Th1 clones showed a significantly increased total number of cells, whereas significantly decreased eosinophils were found in the BALF, when compared with mice with injection of vehicle alone or splenic mononuclear cells. Administration of Th1 clones significantly decreased the infiltration of eosinophils but increased mononuclear cells in the peribronchial area. Goblet cell hyperplasia and peribronchial fibrosis were also suppressed by Th1 clones. The transfer of Th1 cells significantly decreased airway responsiveness. Th1 injection significantly increased interferon gamma in the BALF, but significantly decreased interleukin (IL)-5 and IL-13. Eotaxin mRNA was predominantly expressed in the lungs of asthma model mice, whereas RANTES (regulated on activation, normal T-cell expressed and secreted) predominates in such mice with Th1 transfer. In conclusion, results suggest that the adoptive transfer of T-helper cell type 1 clones can suppress both lung eosinophilia and airway responsiveness, but increase noneosinophilic inflammation in a mouse model of asthma.

Adoptive Transfer↗

Factors causing PAC cake fouling in PAC-MF (powdered activated carbon-microfiltration) water treatment systems.

Two pilot-scale powdered activated carbon-microfiltration (PAC-MF) reactors were operated using river water pretreated by a biofilter. A high permeate flux (4 m/d) was maintained in two reactors with different particle sizes of PAC. High concentration (20 g/L) in the PAC adsorption zone demonstrated 60-80% of organic removal rates. Analysis on the PAC cake fouling demonstrated that attached metal ions play more important role than organic matter attached on PAC to the increase of PAC cake resistance. Effects of factors which may cause PAC cake fouling in PAC-MF process were investigated and evaluated by batch experiments, further revealing that small particulates and metal ions in raw water impose prominent influence on the PAC cake layer formation. Fe (II) precipitates after being oxidized to Fe (III) during PAC adsorption and thus Fe(ll) colloids display more significant effect than other metal ions. At a high flux, PAC cake layer demonstrated a higher resistance with larger PAC due to association among colloids, metals and PAC particles, and easy migration of small particles in raw water into the void space in the PAC cake layer. Larger PAC possesses much more non-uniform particle size distribution and larger void space, making it easier for small colloids to migrate into the voids and for metal ions to associate with PAC particles by bridge effect, hence speeding up and intensifying the of PAC cake fouling on membrane surface.

Adsorption↗

Preoperative concurrent chemoradiotherapy with cisplatin and docetaxel in patients with locally advanced non-small-cell lung cancer.

The objective of this study was to assess the feasibility and effectiveness of an induction chemoradiotherapy regimen followed by surgery in patients with locally advanced non-small-cell lung cancer (LA-NSCLC). A total of 22 patients with LA-NSCLC were treated with induction chemoradiotherapy consisting of cisplatin (40 mg m(-2)) and docetaxel (40 mg m(-2)) given on days 1, 8, 29 and 36 plus concurrent thoracic irradiation at a dose of 40-60 Gy (2 Gy fraction(-1) day(-1)). Surgical resection was performed within 6 weeks after completion of induction therapy. Objective response to the induction therapy was obtained in 16 patients (73%). In all, 20 patients (91%) underwent surgery and complete resection was achieved in 19 patients (86%). Pathological downstaging and pathological complete response were obtained in 14 (64%) and five (23%) patients, respectively. With a median follow-up period of 32 months, the calculated 3-year overall and progression-free survival rates were 66 and 61%, respectively. It is noteworthy that the 3-year overall survival rate in 14 patients achieving pathological downstaging was extremely high (93%). Toxicity was manageable with standard approaches. No treatment-related deaths occurred. This combined modality treatment is feasible and highly effective in patients with LA-NSCLC. The results warrant further large-scale study to confirm the effectiveness of this regimen.

Adult↗

Repressive effects of yeast extract on photoreactivation of Escherichia coli.

Photoreactivation of Escherichia coli K12 (IFO 3301), in the presence or absence of yeast extract (YE), was investigated after inactivation by low-pressure UV lamp. An endonuclease sensitive site (ESS) assay was used to determine the UV-induced pyrimidine dimers in the genome of E. coli, while a colony-forming ability (CFA) test was also used to examine the survival ratio of E. coli. The YE solution reduced the CFA recovery at a final concentration of 125 mg/L. A dialysis of the YE solution indicated that the YE fraction (with nominal molecular weight >1,000 and <3,500) was effective at repressing the CFA recovery. Interestingly, the repair of ESS was equivalent regardless of the presence of the YE dialysate, while the CFA recovery was significantly repressed in the presence of YE. It was, therefore, suggested that YE components, probably with molecular weights of 1,000-3,500, were effective at repressing the CFA recovery of E. coli without affecting the ESS repair during photoreactivation.

Colony Count, Microbial↗

Series of surveys for enteric viruses and indicator organisms in Tokyo Bay after an event of combined sewer overflow.

Combined sewer overflows (CSOs) have been recognised as one of the serious sources of pollution to the water environment during rain events, although field surveys to investigate the effect of their magnitude and duration on receiving waters have been very limited. The fates of enteric viruses (norovirus G1, G2, enteroviruses) and coliforms were determined in the wastewater treatment plant on a fine day and on a rainy day. Not all microorganisms were reduced in the primary treatment, but were reduced in the secondary treatment. Occurrences of enteric viruses and levels of coliforms were surveyed in the receiving coastal area after a CSO event, with the profiles of the enteric viruses in the coastal seawater being almost at the same positive ratio for 4 d after the CSO event.

Data Collection↗

Cryptosporidium monitoring system at a water treatment plant, based on waterborne risk assessment.

The water volume required for daily monitoring of Cryptosporidium (which can statistically ensure an annual risk of infection below 10(-4)), was assessed by evaluating the applicability of the Poisson lognormal (PLN) distribution in microbial risk assessment. PLN showed as good a fit to the observed data as to the negative binomial distribution. From the estimated PLN distributions for the source and finished water, the efficacy of the oocyst removal by the conventional water treatment process was estimated to follow log-normal distribution (median = 3.16 log10, 95% CI = 4.27-2.05 log10). The 365 consecutive negative results of daily monitoring for 180 L of finished water were found to be statistically equivalent to the annual risk of infection below 10(-4). This research also suggested the possibility of applying a qualitative detection method, such as CC-PCR, as a routine monitoring method for the quantitative risk management.

Animals↗

Comparative analysis of chloroplast DNA in Pyrus species: physical map and gene localization.

A physical map of chloroplast DNA (cpDNA) of pear [Pyrus ussuriensis var. hondoensis (Nakai et Kikuchi) Rehder] was constructed using five restriction enzymes, SalI, XhoI, BamHI, SacI and PstI. This information will make it possible to investigate the phylogenetic relationships between Pyrus species. Pear cpDNA was found to be a circular molecule with a total size of about 156 kb in which two inverted repeats of 24.8 kb divide the molecule into small (17 kb) and large (90 kb) single-copy regions. The endonuclease recognition sites in the physical map were determined by single and double digestion of 13 lambda phage clones which covered the entire sequence of the pear cpDNA. Twenty nine genes were localized on the physical map of the pear cpDNA. The structure of pear cpDNA was almost the same in terms of genome size and gene order as that of tobacco cpDNA. RFLP analysis was carried out on cpDNAs from five Pyrus species (Pyrus pyrifolia, Pyrus ussuriensis, Pyrus calleryana, Pyrus elaeagrifolia and Pyrus communis). Two mutations, a recognition-site mutation and a length mutation (deletion), were found only in the cpDNA of P. pyrifolia cultivars. These mutations were localized on the physical map of pear cpDNA. The number of mutations of cpDNA in Pyrus species are small in comparison with those of other angiosperms, suggesting a high degree of genome conservatism in Pyrus species.

Chloroplasts↗

Bacteriophage-host interaction in the enhanced biological phosphate removing activated sludge system.

Bacteriophages were isolated from a laboratory scale enhanced biological phosphate removing (EBPR) activated sludge process, and their host range was examined. Bacterial isolates to host the bacteriophages were isolated from the EBPR activated sludge process. Bacteriophages were eluted from the EBPR activated sludge, enriched by incubation with the bacterial isolates, and then tested for plaque formation on each of the bacterial isolates. Out of 12 bacterial isolates isolated, 4 supported plaque formation. Four bacteriophages were obtained from the plaques. The host range test was conducted with the combination of the bacteriophage isolates and the bacterial isolates. Three of the bacteriophages were found to form plaques on more than one host, and one of them formed plaques on both gram +ve and gram -ve bacterial isolates. Two of the four bacteriophages failed to form plaques on their original bacterial host, indicating the existence of mutation on either both or one of the host and the bacteriophage. This study strongly suggests that bacteriophages are an active part of the activated sludge microbial ecosystem, having very complex interaction with their host bacteria.

Bacteriophages↗