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Biomedical subjects

H Harada

Publications and source records attributed to H Harada.

At least 19 recordsLinked to original sources
PubMed2006

Establishment of immortalized dental follicle cells for generating periodontal ligament in vivo.

The dental follicle is a mesenchymal tissue that surrounds the developing tooth germ. During tooth root formation, periodontal components, viz., cementum, periodontal ligament (PDL), and alveolar bone, are created by dental follicle progenitors. Here, we report the presence of PDL progenitors in mouse dental follicle (MDF) cells. MDF cells were obtained from mouse incisor tooth germs and immortalized by the expression of a mutant human papilloma virus type 16 E6 gene lacking the PDZ-domain-binding motif. MDF cells expressing the mutant E6 gene (MDF( E6-EGFP ) cells) had an extended life span, beyond 150 population doublings (PD). In contrast, normal MDF cells failed to proliferate beyond 10 PD. MDF( E6-EGFP ) cells expressed tendon/ligament phenotype-related genes such as Scleraxis (Scx), growth and differentiation factor-5, EphA4, Six-1, and type I collagen. In addition, the expression of periostin was observed. To elucidate the differentiation capacity of MDF( E6-EGFP ) cells in vivo, the cells were transplanted into severe combined immunodeficiency mice. At 4 weeks, MDF( E6-EGFP ) cell transplants had the capacity to generate a PDL-like tissue that expressed periostin, Scx, and type XII collagen and the fibrillar assembly of type I collagen. Our findings suggest that MDF( E6-EGFP ) cells can act as PDL progenitors, and that these cells may be a useful research tool for studying PDL formation and for developing regeneration therapies.

Alkaline Phosphatase↗
PubMed2006

The influence of physical-chemical and biological factors on the removal of faecal coliform through down-flow hanging sponge (DHS) system treating UASB reactor effluent.

The mechanism of faecal coliform removal in down-flow hanging sponge (DHS) system treating up-flow anaerobic sludge blanket (UASB) reactor effluent was the subject of this study. The results obtained revealed that the most important removal mechanism of faecal coliform in the DHS system is adsorption, followed by predation. Die-off is a relatively minor removal mechanism in the DHS system. The impact of physical and chemical factors such as contact time, effective sponge bulk volume and pH values on faecal coliform removal has been investigated. Increasing the contact time and sponge bulk volume exerted positive effect on the removal of faecal coliform. Changing the pH values between 6.5 and 9.0 did not show significant impact. Reducing sponge pore size from 1.92 to 0.56 mm, increased faecal coliform removal by a value of 1.3 log10.

Adsorption↗
PubMed2006

Technique and assessment of sentinel lymph node biopsy usefulness in laparoscopy-assisted distal gastrectomy.

BACKGROUND: Recently, some studies have suggested that sentinel node biopsy also can be applied to gastric cancer. The authors apply sentinel lymph node biopsy in laparoscopy assisted distal gastrectomy to perform it as safe limited surgery. Limited surgery is a procedure in which the extent of lesion resection and lymph node dissection is reduced. The authors demonstrate that intraoperative diagnosis of lymph node metastasis is useful in this respect. METHODS: The study was conducted with 38 patients (29 men and 9 women) who had a preoperative diagnosis of T1 tumor invasion. The patients had a mean age of 66.2 years. Patent blue (1%) was injected submucosally into four or five different sites around the primary tumor at 1 ml per site. Blue-stained lymphatics and lymph nodes could be seen by turning over the greater omentum and the lesser omentum extraperitoneally. If blue nodes were found, biopsy was performed. RESULTS: The mean number of blue nodes dissected was 2.5 +/- 1.9. Intraoperative identification and biopsy of blue nodes could be performed for 35 (92.1%) of the 38 patients. Of the 35 patients in whom blue nodes were identified, 4 (9.7%) had metastases in blue nodes confirmed by intraoperative frozen-section diagnosis. Intraoperative frozen-section diagnosis was negative for blue node metastasis in 31 patients. Postoperative permanent section diagnosis also showed no evidence of lymph node metastasis in these 31 patients (100% accuracy, 0% false-negative rates). CONCLUSION: The reported method allows observation of blue-stained lymphatics up to 2 h after patent blue injection. Sentinel node biopsy was performed in laparoscopy assisted distal gastrectomy, making it technically equivalent to open gastrectomy. Sentinel node biopsy can serve as a method to determine the appropriate use of laparoscopy assisted distal gastrectomy for management of T1 gastric cancer.

Aged↗
PubMed2006

Development of diffusion-weighted image using a 0.3T open MRI.

The purpose of this study was to develop a new technique for diffusion-weighted MRI (DWI) with a low-field scanner. DWI is becoming important for assessment of acute stroke. Until recently DWI required expensive technology. We developed multishot-DWI sequence for 0.3T open type MR imager. We prospectively studied forty patients on this 0.3T MRI and compared this DWI to single-shot-DWI by 1.5T-MRI. Group A: Twenty-four patients with acute cerebral infarctions detected by 1.5T-DWI were re-examined using 0.3T-DWI within 24 hours. Sixteen patients with acute cerebral infarctions detected by 0.3T-DWI were re-examined using 1.5T-DWI within 24 hours. In 22 (92%) of 24 cases, 0.3T-DWI showed high signal. In the other two patients, motion artifact distorted 0.3T-DWI. Group B: In all 16 patients, all infarctions detected by 0.3T-DWI showed high signal on 1.5T-DWI. These preliminary data show that, as long as the patient is able to keep still, multishot-DWI can be acquired successfully on a 0.3T open type MRI system.

Aged↗
PubMed2006

Brain olfactory activation measured by near-infrared spectroscopy in humans.

In recent years, near-infrared spectroscopy (NIRS) has been used to study functional activation of various areas of the brain. This is based on the assumption that an increase in the recorded oxygenated haemoglobin (HbO2) concentration represents an increase in blood flow, which in turn reflects neuronal activation. The aim of this preliminary study was to use NIRS to monitor the activity of the olfactory cortex, as mirrored by the haemodynamic response, when subjects were exposed to olfactory stimuli. A NIRO 300 (Hamamatsu Photonics, Hamamatsu, Japan) device was used. The optodes were placed on the right forehead and right temporal, parietal and occipital regions. Changes in the concentration of HbO2 and deoxygenated haemoglobin during olfactory stimulation were monitored. Olfactory stimulation was performed with vanilla essence, strawberry essence and scatol. During olfactory stimulation, cerebral HbO2 concentration increased over the frontal region. However, in the temporal, parietal and occipital regions, little or no HbO2 changes were recorded. This study shows that human brain cortical activation following olfactory stimulation can be recorded by NIRS. This NIRS analysis may therefore provide the basis for future development of an objective olfactory test in humans.

Adult↗
PubMed2006

Hyperactivation of the RAS signaling pathway in myelodysplastic syndrome with AML1/RUNX1 point mutations.

AML1/RUNX1 mutations have been reported frequently in myelodysplastic syndrome (MDS) patients, especially those diagnosed with refractory anemia with excess blast (RAEB), RAEB in transformation (RAEBt), or AML following MDS (these categories are defined as MDS/AML). Although AML1 mutations are suspected to play a pivotal role in the development of MDS/AML, acquisition of additional genetic alterations is also necessary. We analyzed gene alterations in MDS/AML patients with AML1 mutations, comparing them to alterations in those without an AML1 mutation. AML1 mutations were significantly associated with -7/7q-, whereas MDS/AML patients without AML1 mutations showed a high frequency of -5/5q- and a complex karyotype. Patients with AML1 mutations showed more mutations of their FLT3, N-RAS, PTPN11, and NF1 genes, resulting in a significantly higher mutation frequency for receptor tyrosine kinase (RTK)-RAS signaling pathways in AML1-mutated MDS/AML patients compared to AML1-wild-type MDS/AML patients (38% versus 6.3%, P < 0.0001). Conversely, p53 mutations were detected only in patients without AML1 mutations. Furthermore, blast cells of the AML1-mutated patients expressing surface c-KIT, and SHP-2 mutants contributed to prolonged and enhanced extracellular signal-regulated kinase activation following stem cell factor stimulation. Our results suggest that MDS/AML arising from AML1/RUNX1 mutations has a significant association with -7/7q- alteration, and frequently involves RTK-RAS signaling pathway activation.

Adult↗
PubMed2006

On-site treatment of high-strength alcohol distillery wastewater by a pilot-scale thermophilic multi-staged UASB (MS-UASB) reactor.

A pilot-scale multi-staged UASB (MS-UASB) reactor with a working volume of 2.5 m3 was operated for thermophilic (55 degrees C) treatment of an alcohol distillery wastewater for a period of over 600 days. The reactor steadily achieved a super-high rate COD removal, that is, 60 kgCOD m(-3) d(-1) with over 80% COD removal. However, when higher organic loading rates were further imposed upon the reactor, that is, above 90 kgCOD m(-3) d(-1) for barely-based alcohol distillery wastewater (ADWW) and above 100 kgCOD m(-3) d(-1) for sweet potato-based ADWW, the reactor performance somewhat deteriorated to 60 and 70% COD removal, respectively. Methanogenic activity (MA) of the retained sludge in the thermophilic MS-UASB reactor was assessed along the time course of continuous run by serum-vial test using different substrates as a vial sole substrate. With the elapsed time of continuous run, hydrogen-utilizing MA, acetate-utilizing MA and propionate-fed MA increased at maximum of 13.2, 1.95 and 0.263 kgCOD kgVSS(-1) d(-1), respectively, indicating that propionate-fed MA attained only 1/50 of hydrogen-utilizing MA and 1/7 of acetate-utilizing MA. Since the ADWW applied herewith is a typical seasonal campaign wastewater, the influence of shut-down upon the decline in sludge MA was also investigated. Hydrogen-utilizing MA and acetate-utilizing MA decreased slightly by 3/4, during a month of non-feeding period, whereas propionate-fed MA were decreased significantly by 1/5. Relatively low values of propionate-fed MA and its vulnerability to adverse conditions suggests that the propionate degradation step is the most critical bottleneck of overall anaerobic degradation of organic matters under thermophilic condition.

Alcohols↗
PubMed2006

Combining UASB and the "fourth generation" down-flow hanging sponge reactor for municipal wastewater treatment.

A "fourth generation" down-flow hanging sponge (DHS) Reactor has been developed and proposed as an improved variant of post-treatment system for UASB treating domestic wastewater. This paper evaluates the potential of the proposed combination of UASB and DHS as a sewage treatment system, especially for developing countries. A pilot-scale UASB (1.15 m3) and DHS (0.38 m3; volume of sponge) was installed in a municipal sewage treatment site and constantly monitored for 2 years. UASB was operated at an HRT of 6 h corresponding to an organic load of 2.15 kg-COD/m3 per day. Subsequently, the organic load in DHS was 2.35 kg-COD/m3 per day, operated at an HRT of 2 h. Organic removal by the whole system was satisfactory, accomplishing 96% of unfiltered BOD removal and 91% of unfiltered COD removal. However, nitrification decreased from 56% during the startup period to 28% afterwards. Investigation on DHS sludge was made by quantifying it and evaluating oxygen uptake rates with various substrates. Average concentration of trapped biomass was 26 g-VSS/L of sponge volume, increasing the SRT of the system to 100-125 d. Removal of coliforms obtained was 3-4 log10 with the final count of 10(3) to 10(4) MPN/100 ml in DHS effluent.

Biomass↗
PubMed2006

Anaerobic digestion with partial ozonation minimises greenhouse gas emission from sludge treatment and disposal.

A novel anaerobic digestion process combined with partial ozonation on digested sludge was demonstrated for improving sludge digestion and biogas recovery by full-scale testing for 2 years and its performance was compared with a simultaneously operated conventional anaerobic digestion process. The novel process requires two essential modifications, which are ozonation for enhancing the biological degradability of sludge organics and concentrating of solids in the digester through a solid/liquid separation for extension of SRT. These modifications resulted in high VSS degradation efficiency of ca. 88%, as much as 1.3 times of methane production and more than 70% reduction in dewatered sludge cake production. Based on the performance, its energy demands and contribution for minimisation of greenhouse gas emission was evaluated throughout an entire study of sludge treatment and disposal schemes in a municipality for 130,000 p.e. The analysis indicated that the novel process with power generation from biogas would lead to minimal greenhouse gas emission because the extra energy production from the scheme was expected to cover all of the energy demand for the plant operation, and the remarkable reduction in dewatered sludge cake volumes makes it possible to reduce N2O discharge and consumption of fossil fuel in the subsequent sludge incineration processes.

Algorithms↗
PubMed2006

Real-time PCR quantification of nitrite reductase (nirS) genes in a nitrogen removing fluidized bed reactor.

Molecular approaches were applied to identify and enumerate denitrifying bacteria subsisting in a fluidized bed reactor (FBR). The FBR was continuously operated as a unit for the removal of nitrogen from the effluents of domestic sewage treatment plant, with an additional supply of methanol as a carbon source. By denaturing gradient gel electrophoresis (DGGE) and sequence analysis of 16S ribosomal RNA genes, Thauera group was found to be dominant among the denitrifying bacteria in the FBR sludge. Oligonucleotide probe THA155 for fluorescence in situ hybridization (FISH) was newly designed for specifically targeting the Thauera group. However, the THA155 signal obtained from the sludge was only 0.9-5.7% of the DAPI-stained total cells. The real-time polymerase chain reaction (PCR) targeting the sequences of nitrite reductase (NIR) gene, a key enzyme of denitrification processes, was performed to quantify the cells of denitrifying bacteria cells including the Thauera group in FBR sludge. An excellent correlation was obtained between the numbers of nirS genes and the activity of denitrifiers in the FBR sludge.

Bacteria↗
PubMed2006

Low strength wastewater treatment under low temperature conditions by a novel sulfur redox action process.

The objective of this research is to make a novel wastewater treatment process activated by a sulfur-redox cycle action of microbes in low temperature conditions. This action is carried out by sulfate-reducing bacteria (SRB) and sulfur-oxidizing bacteria (SOB). The process was comprised of a UASB reactor as pre-treatment and an aerobic downflow hanging sponge (DHS) reactor as post-treatment. As the results of reactor operation, the whole process achieved that over 90% of CODcr removal efficiency, less than 30 mgCODcr/L (less than 15 mgBOD/L) of final effluent, at 12 h of HRT and at 8 degrees C of UASB reactor temperature. Acetobacterium sp. was detected as the predominant species by PCR-DGGE method targeting 16SrDNA with band excision and sequence analysis. In the UASB reactor, various species of sulfate-reducing bacterium, Desulfobulbus sp., Desulfovibrio sp., and Desulfomicrobium sp., were found by cloning analysis. In the DHS reactor, Tetracoccus sp. presented as dominant. The proposed sulfur-redox action process was considered as an applicable process for low strength wastewater treatment in low temperature conditions.

Bioreactors↗
PubMed2006

Effect of inorganic carbon on nitrite accumulation in an aerobic granule reactor.

Pilot scale experiments were performed to evaluate the potential of nitrite type nitrification process with an airlift reactor and granular biomass. Initially, oxygen limitation was used as the main control parameter for accumulating nitrite in the effluent. After 30 d operation, the maximum nitrite conversion rate reached 2.5 kgNO2-N m(-3) d(-1), average diameter of the granule was 0.7 mm. Nitrite type reaction continued over 100 d, but nitrate formation increased after 150 d of operation. Once nitrate formation increased, oxygen limitation could not eliminate nitrite oxidising bacteria from granule. To overcome nitrate formation, laboratory scale batch experiments were conducted and it revealed a high concentration of inorganic carbon which had a significant effect on nitrite accumulation. Following this new concept, inorganic carbon was fed to the pilot scale reactor by changing pH adjustment reagent from NaOH to Na2CO3 and nitrite accumulation was recovered successfully without changing DO concentration. These results show that a high concentration of inorganic carbon is one of the control parameters for accumulating nitrite in biofilm nitrification system.

Aerobiosis↗
PubMed2006

Predicting struvite formation for phosphorus recovery from human urine using an equilibrium model.

Interest in phosphorus recovery from urine diverted from faeces has been growing recently. Phosphorus in urine can be precipitated out as struvite (MgNH4PO4 x 6H2O) with addition of magnesium salt under alkaline conditions. Struvite formation, however, should be more well understood for its practical application. We predicted the struvite formation with a development of a new equilibrium model. The model considered the formation of eight different kinds of precipitates, including struvite, with effects of ionic strength and temperature. In addition, experiments on struvite formation in urine were conducted for the model validation. The model prediction of struvite formation had good agreement with the experimental results. The optimum pH to form struvite was predicted to be 9.4-9.7. In order to precipitate 99% of phosphate in urine with 1.5 fold Mg concentration to PO4-P, the pH value was necessary to be more than 8.1 based on the model prediction.

Humans↗
PubMed2006

Cyclin B1 is useful to predict occult cervical lymph node metastases in tongue carcinoma.

Cyclin B1 plays an important role in regulating the G2-M transition of the cell cycle. In this study, we evaluated the immunohistochemical expression of cyclin B1 in a series of 40 oral tongue squamous cell carcinomas and analyzed the findings in relation to both clinicopathological variables and the Ki-67 labeling index. In normal squamous epithelium, cyclin B1 was localized in the nucleus and was expressed only in several cells of the basal and parabasal layers. By contrast, in dysplastic epithelium, cyclin B1 was localized in the cytoplasm and nuclei, and the numbers of cyclin B1 positive cells increased compared with normal epithelium. In tumor tissues, cyclin B1 expression was observed mainly in the cytoplasm, and the cyclin B1 labeling index ranged from 2 to 21%, with a mean of 11%. Cyclin B1 overexpression was positively correlated with occult cervical lymph node metastases and the number of mitotic cells. In addition, there was a positive relationship between labeling indices of cyclin B1 and Ki-67. These findings indicate that cyclin B1 could be a useful prognostic marker of occult cervical lymph node metastases in tongue carcinoma.

Adult↗
PubMed2005

Peroxiredoxin I expression in tongue squamous cell carcinomas as involved in tumor recurrence.

Peroxiredoxin (Prx) I is an antioxidant protein expressed in proliferating cells. We investigated Prx I as marker for tongue cancer status by correlating clinical features with Prx I expression. Samples from 132 patients with squamous cell carcinoma in the tongue were examined by immunohistochemistry with an anti-Prx I antibody. Correlations between Prx I expression and the clinical features of tumors were statistically determined using univariate and multivariate analyses. Univariate analysis showed Prx I was significantly associated with local recurrence (P=0.033). By multiple logistic regression analysis, Prx I expression was associated with local recurrence (odds ratio: 2.84; 95% confidence interval: 1.09-7.43; P=0.034) and lymph node recurrence (odds ratio: 2.86; 95% confidence interval: 1.02-8.01; P=0.046). Our results suggested that Prx I expression indicates tumors with a high potential for recurrence. Prx I may be used clinically to guide treatment for squamous cell carcinoma of the tongue.

Adult↗
PubMed2005

Treatment of wastewater containing a large amount of suspended solids by a novel multi-staged UASB reactor.

Treatment of artificial wastewater containing a large amount of suspended solids comprised of soybean processing waste and pig fodder was studied using a novel multi-staged upflow anaerobic sludge blanket reactor. The reactor consisted of three compartments, each containing a gas solid separator. The wastewater had chemical oxygen demand of approximately 21600 mg l(-1), suspended solids of 12800 mg l(-1), and an ammonia concentration of 945 mg l(-1). A continuous experiment without effluent circulation showed that the multi-staged reactor was not that effective for the treatment of wastewater containing a large amount of suspended solids. However, operation of the reactor with circulation of effluent enabled the reactor to achieve organic removal of 85% and approximately 70% methane conversion at loading rates of between 4.0 to 5.4 kg-chemical oxygen demand per cubic meter per day, meaning that the reactor was more effective when effluent was circulated. Morphological investigation revealed that the crude fiber in the sludge was partially degraded and that it had many small depressions on its surface. Evolved biogas may have become caught in these depressions of the fibers and caused washout of the sludge.

Anaerobiosis↗
PubMed2005

Hydrolytic activity of alpha-amylase in anaerobic digested sludge.

Hydrolysis is usually considered to be a rate-limiting step in anaerobic digestion. For improving anaerobic solid waste treatments, it is essential to elucidate the mechanism of hydrolysis. In this study, alpha-amylase, one of the hydrolytic enzymes, was investigated for the elucidation of more precise mechanism of hydrolysis. Alpha-amylase activity of solid starch-degrading bacteria (SDB) was estimated through batch experiments with several different substrates and with distinction between cell-bound and cell-free alpha-amylase. Monitoring of newly isolated strains of SDB was done by fluorescence in situ hybridization. Results indicated that cell-bound alpha-amylase is chiefly responsible for the hydrolysis in the digested sludge, providing very useful information that the contact between microbial cells and solids is significantly important. The activity of alpha-amylase of the digested sludge remained quite low when not required, but increased as they recognized appropriate substrates. Several-fold higher activity was obtained for starch or maltose as compared to glucose only.

Aeromonas↗