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Biomedical subjects

C Kobayashi

Publications and source records attributed to C Kobayashi.

At least 19 recordsLinked to original sources

Similar acute molecular responses to equivalent volumes of isometric, lengthening, or shortening mode resistance exercise.

The present study was undertaken to test the hypothesis that the contraction mode of action [static-isometric (Iso), shortening-concentric (Con), or lengthening-eccentric (Ecc)] used to stress the muscle provides a differential mechanical stimulus eliciting greater or lesser degrees of anabolic response at the initiation of a resistance training program. We performed an acute resistance training study in which different groups of rodents completed four training sessions in either the Iso, Con, or Ecc mode of contraction under conditions of activation and movement specifically designed to elicit equivalent volumes of force accumulation. The results of this experiment indicate that the three modes of contraction produced nearly identical cell signaling, indicative of an anabolic response involving factors such as increased levels of mRNA for IGF-I, procollagen III alpha1, decreased myostatin mRNA, and increased total RNA concentration. The resulting profiles collectively provide evidence that pure mode of muscle action, in and of itself, does not appear to be a primary variable in determining the efficacy of increased loading paradigms with regard to the initiation of selected muscle anabolic responses.

Animals↗

An ex vivo evaluation of a new root canal irrigation technique with intracanal aspiration.

AIM: To evaluate the effectiveness of a new root canal irrigation technique with intracanal aspiration in removing the smear layer and to assess irrigant extrusion ex vivo. METHODOLOGY: Thirty-five instrumented canals of extracted human canine teeth that had been resected apically by removing 3 mm of the root tip were divided into one control and four experimental groups of seven teeth each. The roots were fixed in a plastic case and surrounded with normal saline agar coloured with 1% acid red. No irrigation was performed in the control teeth. Each root canal in the experimental groups was irrigated with 9 mL of 14% ethylenediaminetetraacetic acid for 3 min, and then with 6 mL of 6% sodium hypochlorite (NaOCl) for 2 min. In the intracanal aspiration technique, the irrigant was delivered from the tip of an injection needle placed 12 mm from the apical root-end and an aspiration needle that was connected to a Root ZX apex locator placed 2 and 3 mm short of the apical root-end in groups 1 and 2, respectively. In the conventional method, the tip of an injection needle used for delivery of the irrigant and as an active electrode was placed 2 and 3 mm short of the apical root-end in groups 3 and 4, respectively, the tip of the aspiration needle was placed 12 mm from the apical root-end in these groups. The readings of the Root ZX during irrigation were recorded. The cleanliness of the canal was evaluated by scoring smear layer from scanning electron microscopy (SEM) images of the canal. Extrusion of NaOCl was detected by measuring the discoloured area of the agar around the apical root-end. The data obtained were statistically analysed by one-way anova, the Kruskal-Wallis test and Friedman's test. RESULTS: In the SEM study, the canals in groups 1-3 were significantly cleaner than those in the control and group 4 (P < 0.05). The mean Root ZX readings in groups 1-3 were approximately "0.5". The discoloured area in group 3 was significantly larger than the other groups (P < 0.05). CONCLUSIONS: Irrigation using the intracanal aspiration technique allowed more effective removal of the smear layer than that performed by the conventional method in an apically resected canine tooth. The intracanal aspiration technique produced limited extrusion of the irrigant beyond the apical foramen.

Apicoectomy↗

The effects of file size, sodium hypochlorite and blood on the accuracy of Root ZX apex locator in enlarged root canals: an in vitro study.

BACKGROUND: The initial electronic apex locator (EAL) length measurement is generally established with a small-sized file. It is not known whether file size would be interfering with the reading accuracy of the EAL. This study aimed to evaluate the effect of file size on the accuracy of Root ZX apex locator using an agar model when sodium hypochlorite solution or blood was present during electronic measurements in enlarged root canals. METHODS: A total of 36 extracted lower premolars were used. In stage 1, the canals were instrumented using size 10-40 K-files with a size 40 K-file as the master apical file (MAF). The teeth were then divided randomly into two groups of 18 teeth each. In group A, the teeth were mounted in one per cent agar and irrigated with six per cent sodium hypochlorite solution (NaOCl), while in group B the teeth were mounted in agar and irrigated with human blood. In stage 2, the canals were enlarged using a size 60 K-file as the MAF. In stages 1 and 2, the apical portions of the canals were instrumented using the step-back sequence (up to a size 80 K-file). In stage 3, the canals were enlarged using a size 80 K-file as the MAF. In each stage, the length was measured with a Root ZX until the meter value reached 'APEX' using small and large size files. RESULTS: Three-way ANOVA and Bonferroni test showed that file size, stage of preparation and type of irrigant all had a significant influence on the measurement error (P < 0.0001), with all the interactions between these three factors being significant (P < 0.0001). CONCLUSIONS: As the diameter of the root canal increased, the measured length with the smaller size files became shorter. A file of a size close to the prepared canal diameter should be used for root length measurement in the presence of blood. In the presence of NaOCl, the Root ZX was highly accurate even when the file was much smaller than the diameter of the canal. The agar model was effective and suitable for testing EALs in vitro.

Analysis of Variance↗

Selective and efficient culturing of retinal pigment epithelial cells using a feeder layer.

BACKGROUND: The techniques to isolate and purify retinal pigment epithelial (RPE) cells from small piece of autologous tissues are extremely difficult, and it is important to develop an efficient cell culture technique for RPE cells. The purpose of this study was to investigate the effect of 3T3-J2 cells and conditioned medium from 3T3-J2 cells on the proliferation of cultured RPE cells. METHODS: RPE cells from pigmented rabbits and a human RPE-derived cell line, ARPE-19, were used. First, the effects of co-culturing RPE cells with 3T3-J2 cells on the growth of the cells were analyzed. Second, the effects of the conditioned medium from 3T3-J2 cells on the proliferation of both types of cells were investigated. And third, the effects of the conditioned medium on RPE cell culture from a surgically removed choroidal neovascular (CNV) membrane were investigated. RESULTS: The 3T3-J2 cells increased the proliferation of both rabbit RPE cells and ARPE-19 cells. The number of rabbit RPE cells cultured in a mixture of the conditioned medium from 3T3-J2 cells was significantly higher than that in the reported optimal condition, and a similar tendency was observed for ARPE-19 cells. The results from enzyme-linked immunosorbent assay showed the presence of PDGF-AB, VEGF and IGF-I in the conditioned medium. The conditioned medium also promoted selective growth of human RPE cells from CNV. DISCUSSION: The results from this study present the conditions for efficient and selective culture of primary RPE cells.

3T3 Cells↗

Analysis of forces developed during mechanical preparation of extracted teeth using RaCe rotary instruments and ProFiles.

AIM: To compare the torque and load during instrumentation with ProFile and RaCe nickel-titanium rotary instruments. METHODOLOGY: Thirty human incisor roots 12 mm in length were embedded in epoxy resin, and divided into three groups of 10 specimens each. Instruments employed in each group were as follows: RaCe .02 Step Back Files in group 1; RaCe .04 Step Back Files in group 2; and .04 ProFiles in group 3. Two load cells, whose outputs were connected to a digital oscilloscope, recorded torque and vertical load. Data were analysed by one-way anova. RESULTS: Torque values were statistically different between groups 1 and 3, and between groups 2 and 3 (torque was higher in group 3 than in groups 1 and 2). Vertical loads were statistically higher in group 3 than in groups 1 and 2. CONCLUSIONS: Torsional and vertical forces can be evaluated during instrumentation of straight root canals using the device and methods described in this study. When the step-back technique was employed, torque and vertical load obtained with RaCe instruments were lower than that obtained with ProFiles.

Analysis of Variance↗

Detection of the second mesiobuccal canal in mesiobuccal roots of maxillary molar teeth ex vivo.

AIM: To assess the effectiveness of magnification and dentine removal (troughing) when locating the second mesiobuccal canal in mesiobuccal roots of maxillary molars. METHODOLOGY: A total of 208 extracted human maxillary molars were examined. After crown and pulp removal, the MB1 and 2 canals in the mesiobuccal root were located in three stages that were performed by two undergraduate dental students. Stage 1: canals were located with an endodontic explorer; stage 2: additional canals in the same teeth were located under magnification with a digital microscope (VH-8000, Keyence, Japan); stage 3: additional canals in the same teeth were located by removing dentine (troughing) from the pulp chamber floor within 3 mm from MB1 canal towards the palatal canal with an Enac ultrasonic tip (ST 21, Osada, Japan). In each group, the canals were prepared with Gates Glidden burs and K-files. The distal and palatal roots were then removed, and Indian ink was injected into the canal system within the mesio-buccal root. The root surfaces were washed with 6% NaOCl, and then rendered transparent to observe canal morphology. The root canal configurations were classified into five categories following the modified Weine's classification. RESULTS: More than one canal in the mesio-buccal root was observed in 48% of specimens. Detection rates of multiple canals were 7, 18 and 42% following stages 1, 2 and 3, respectively. There was a significant difference between the stages for detecting the MB2 canal (P < 0.05, Friedman test). CONCLUSIONS: Both magnification (stage 2) and dentine removal under magnification (stage 3) were effective in detecting the presence of the MB2 canal. However, MB2 canals could not be detected in 13% of the teeth because of canal calcification or branching located more apically.

Dental Pulp Cavity↗

Intercalary regeneration in planarians.

How can a planarian regenerate its entire body from a small portion of its body? Neoblasts, the totipotent stem cells of planarian, are assumed to be able to produce all missing cell types. However, we do not know how the cell fate of these cells is controlled during regeneration. Our recent studies with molecular markers suggest that intercalary regeneration is the fundamental principle in planarian regeneration. Here, we introduce the intercalation induced by ectopic grafting along the anteroposterior (A-P), dorsoventral (D-V), and left-right (L-R) axes. Blastema formation is evoked by ectopic D-V interactions after wound closure. Intercalation between the blastema and stump induces rearrangement of the positional identities along the A-P axis. Consequently, totipotent stem cells change their differentiation patterns according to the newly rearranged positional identities along the A-P, D-V, and L-R axes. According to the classic view, the blastema is regarded as the place where undifferentiated cells accumulate and regenerative events occur. Here, we propose a new interpretation, i.e., that the blastema may work as a signaling center inducing intercalary regeneration. Also, the roles of molecules and genes involved in intercalary regeneration are discussed.

Animals↗

Evaluation of root-end cavity preparation using ultrasonic retrotips.

AIM: To evaluate and compare the efficiency of root-end preparations using ultrasonic retrotips coated with diamond and zirconium nitride. METHODOLOGY: Eighty-five extracted single-rooted teeth were root filled, and then resected 3 mm from their apices. Root-end cavities were prepared with KiS (zirconium nitride-coated retrotip), CT-5 (stainless steel tip) or diamond-coated (DC) ultrasonic retrotips, and 10 teeth served as controls. Thirty teeth were used for evaluation of the time required to prepare the root-end cavity, the number of microcracks produced on the resected surface and the number of dentinal tubule openings on the root-canal wall using scanning electron microscope (SEM) images. A further 55 teeth were used for evaluation of dye penetration following filling of the root-end cavities with Super EBA. The degree of dye penetration in millimetres was measured under the microscope after 7 days of immersion in India ink. Statistical analyses were performed using the one-way ANOVA and Scheffe's F-test as the post hoc test. RESULTS: There was no significant difference in the number of microcracks and dentinal tubule openings present in the root apices prepared by the three retrotips. The time required for root-end cavity preparation using the DC retrotip was significantly less than that using the other groups (P<0.01). Positive controls showed dye penetration throughout the length of the root-end cavity, and negative controls showed no dye penetration. There was no significant difference between the three experimental groups in dye penetration. CONCLUSIONS: In this laboratory study, the time required to prepare root-end cavities using KiS retrotips was the same as that using CT-5 retrotips, and longer than that using DC retrotips. There was no significant difference in the number of microcracks or dye penetration between the three kinds of retrotips.

Analysis of Variance↗

Achieving recommended goals for blood-glucose and blood-pressure control in diabetic patients: a multi-center cross-sectional evaluation in the Niigata prefecture.

Hypertension in patients with diabetes mellitus can contribute to a deterioration of the patient's status as much as most diabetes mellitus-related complications. Previous interventional trials have found that to prevent the onset and progression of diabetic microangiopathy, the optimal glycemic threshold is an HbA1c of less than 6.5% and the optimal blood pressure is less than 130/85 mmHg. The present study investigated the prevalence of achieving these recommended goals for glycemic and blood pressure control in type 2 diabetic patients in the Niigata prefecture. A questionnaire was administered in this multi-center study to 3573 patients from 92 participating Niigata Diabetic Complication Study hospitals. Patients aged 63.5 +/- 11 years with type 2 diabetes were evaluated. Only 46.8% of patients achieved the recommended glycemic control level, and only 41.4% of patients achieved the BP target. Moreover, only 20.5% of all patients achieved both target levels for blood glucose and blood pressure. This demonstrates the difficulty in clinical practice of achieving concurrent glucose and blood pressure control in diabetic patients.

Blood Glucose↗

A scanning electron microscopic study of dentinal erosion by final irrigation with EDTA and NaOCl solutions.

AIM: The purpose of this in vitro study was to examine dentinal erosion caused by final irrigation with EDTA and NaOCl. METHODOLOGY: Twenty-five single-rooted human teeth were instrumented with rotary nickel-titanium Series 29 Profile Instruments. The teeth were divided into five groups and subjected to final irrigation as follows: group A, irrigated with 6% NaOCl (3 mL) for 2 min; group B, 15% EDTA (3 mL) for 1 min; group C, 15% EDTA (3 mL) for 1 min, followed by 6% NaOCl (3 mL) for 2 min; group D, 15% EDTA (3 mL) for 3 min and group E, 15% EDTA (3 mL) for 3 min, followed by 6% NaOCl (3 mL) for 2 min. Photomicrographs of dentinal walls were produced using a scanning electron microscope (3000 x) at 1, 3 and 6 mm from the apex. The amount of debris and dentinal tubule diameter were evaluated, and values were statistically analysed using one-way ANOVA and Fisher's PLSD test. RESULTS: When the root canal was irrigated with 15% EDTA alone, the dentine had a smooth and plane appearance, and dentinal tubule orifices were regular and separated. When the root canal was irrigated with EDTA followed by NaOCL the dentine was eroded and the dentinal tubule orifices were irregular and rough. Dentinal tubule diameter increased to 3.43 +/- 0.23 microm in group C and to 3.93 +/- 0.44 microm in group E. Significant differences were observed between groups B and C, and between groups D and E (P < 0.05). However, more debris was removed by irrigation with EDTA followed by NaOCl than with EDTA alone (P < 0.05). CONCLUSIONS: Final irrigation with 6% NaOCl accelerates dentinal erosion following treatment with 15% EDTA.

Analysis of Variance↗

Expression and characterization of hypoxia-inducible factor (HIF)-3alpha in human kidney: suppression of HIF-mediated gene expression by HIF-3alpha.

Hypoxia inducible factors (HIFs) are heterodimeric transcription factors that regulate a number of adaptive responses to low oxygen tension. They are composed of alpha- and beta-subunits that belong to the basic helix-loop-helix-PAS superfamily. Here we examined the expression of HIF-alpha subunit proteins in the human kidney and found that, in addition to HIF-1alpha and -2alpha, HIF-3alpha was also expressed. The sequence analysis revealed that, like mouse HIF-3alpha, human HIF-3alpha has high similarity with HIF-1alpha and -2alpha in the bHLH and PAS domains, but lacks structures for transactivation found in the C-terminus of HIF-1alpha and -2alpha. Furthermore, we performed reporter gene analysis and showed that HIF-3alpha suppresses hypoxia-inducible HIF-mediated gene expression. HIF-3alpha might be a negative regulator of hypoxia-inducible gene expression in the human kidney.

Amino Acid Sequence↗

Contributory metabolic factors in the development of nephrolithiasis in patients with medullary sponge kidney.

Whether specific metabolic abnormalities are related to nephrolithiasis in patients with medullary sponge kidney (MSK) remains a debated issue. The purpose of this study is to determine metabolic disorders in patients with MSK and nephrolithiasis compared with idiopathic calcium-stone-forming patients. One hundred eighty-four patients with recurrent calcium-stone formations were investigated with regard to metabolic abnormalities. Of these, 22 patients (11.9%; 13 men, 9 women) showed MSK by radiological examination. MSK was defined as a kidney that presented at least three linear or round papillary opacities in the affected papilla on urography. Multiple stones (more than five) existed in both kidneys in all patients with MSK. The remaining 162 patients (109 men, 53 women) were idiopathic calcium-stone formers. Frequencies of low urine volume (urine < 1,500 mL/24 h) and hyperoxaluria (oxalate > 40 mg/24 h) were similar between the groups. Hypercalciuria (men, calcium > 300 mg/24 h; women, calcium of 250 mg/24 h) was found less frequently in the MSK group. The frequency of hypocitraturia (citrate < 300 mg/24 h) was significantly greater in the MSK group than the idiopathic group (77.3% versus 33.9%, respectively). Mean 24-hour urinary excretions of calcium, citrate, uric acid, and magnesium were significantly less in the MSK group. No differences were found in serum calcium, phosphate, and parathyroid hormone levels between the groups. Low urinary excretions of citrate and magnesium are the most typical metabolic disorders that distinguish MSK stone patients from idiopathic calcium-stone-forming patients. In addition to such anatomic abnormalities as ectatic collecting ducts, low levels of urinary inhibitors of stones seem to contribute to the pathogenesis of nephrolithiasis in patients with MSK.

Calcium↗

Benefits of ureteroscopic pneumatic lithotripsy for the treatment of impacted ureteral stones.

PURPOSE: Although new forms of lithotripsy, such as new-generation shockwave and laser lithotripsies, have recently been developed, the optimal treatment for impacted ureteral stones (IUS) remains controversial. We investigated, retrospectively, the outcome of IUS treated with ureteroscopic pneumatic lithotripsy. PATIENTS AND METHODS: Twenty-two IUS cases were treated using ureteroscopic pneumatic lithotripsy. Of 22 stones, 8 (36%) were treated by ureteroscopic pneumatic lithotripsy as initial treatment and 14 (64%) with pneumatic lithotripsy as an auxiliary treatment after SWL. The stone sizes ranged from 7 to 16 mm with 14 located in the proximal, 3 in the middle, and 5 in the distal ureter. RESULTS: Twenty stones (91%) were effectively fragmented by ureteroscopic pneumatic lithotripsy and eliminated within 1 month after treatment. One stone could not be observed with the ureteroscope secondary to the kinking and stricture of the ureter under the stone, and the stone moved into the kidney during the procedure. The stone was subsequently treated successfully with SWL. One other stone was first fragmented into two pieces; and one big piece, which migrated into the kidney, was treated successfully with SWL. Complications such as bleeding, ureteral injury, and perforation did not occur. Although a few small fragments migrated into the kidney during the procedure in three cases, the overall stone-free rate at 1 month after a one-session treatment with pneumatic lithotripsy was 91%. CONCLUSION: Impacted ureteral stones can be treated effectively with ureteroscopic pneumatic lithotripsy in both the short and the long term.

Adult↗

Emx2 directs the development of diencephalon in cooperation with Otx2.

The vertebrate brain is among the most complex biological structures of which the organization remains unclear. Increasing numbers of studies have accumulated on the molecular basis of midbrain/hindbrain development, yet relatively little is known about forebrain organization. Nested expression among Otx and Emx genes has implicated their roles in rostral brain regionalization, but single mutant phenotypes of these genes have not provided sufficient information. In order to genetically determine the interaction between Emx and Otx genes in forebrain development, we have examined Emx2(-/-)Otx2(+/-) double mutants and Emx2 knock-in mutants into the Otx2 locus (Otx2(+/Emx2)). Emx2(-/-)Otx2(+/-) double mutants did not develop diencephalic structures such as ventral thalamus, dorsal thalamus/epithalamus and anterior pretectum. The defects were attributed to the loss of the Emx2-positive region at the three- to four-somite stage, when its expression occurs in the laterocaudal forebrain primordia. Ventral structures such as the hypothalamus, mammillary region and tegmentum developed normally. Moreover, dorsally the posterior pretectum and posterior commissure were also present in the double mutants. In contrast, Otx2(+/Emx2) knock-in mutants displayed the majority of these diencephalic structures; however, the posterior pretectum and posterior commissure were specifically absent. Consequently, development of the dorsal and ventral thalamus and anterior pretectum requires cooperation between Emx2 and Otx2, whereas Emx2 expression is incompatible with development of the commissural region of the pretectum.

Animals↗

[Swallowing rehabilitation in two elderly patients with cerebral infarction].

We report two cases of cerebral infarction in which swallowing function improved following swallowing rehabilitation. Patient 1 was an 82-year-old man, who was admitted due to rheumatoid arthritis and multiple cerebral infarction, suffering from aspiration pneumonia. The abnormality of swallowing was assessed by the water swallowing test and videofluorography. It has been reported that videofluorography is useful in the diagnosis of aspiration. Three weeks after the start of swallowing rehabilitation, the serum level of inflammatory markers and the chest X-ray had returned to normal. His score on the water swallowing test had improved. Patient 2 was a 68-year-old [correction of 62] man, who was admitted with severe hemiplegia, dysphagia and dysarthria. One month after the swallowing rehabilitation, videofluorography showed that the magnitude of aspiration into the trachea had decreased and the pooling of barium in the piriform sinus had disappeared. The patient could begin taking a little food by mouth. These results suggest that swallowing rehabilitation will be affect the clinical improvement of swallowing function and help preventing aspiration pneumonia in our hospital.

Activities of Daily Living↗

[The effects of pulmonary rehabilitation with chronic obstructive pulmonary disease (COPD)].

It is controversial whether pulmonary rehabilitation is effective in patients with chronic obstructive pulmonary disease (COPD). To test the effect of pulmonary rehabilitation, 7 patients with COPD (aged 76.0 +/- 2.6 years) were enrolled in pulmonary rehabilitation program for 6 weeks. The program consisted of relaxation, pursed lip breathing, diaphragmatic breathing, panic control, muscle stretch gymnastics, and exercise training. The distance of the 6-minute walking test increased significantly from 246.4 +/- 38.0 (m) to 304.3 +/- 28.4 (m) (p < 0.05). The minimum SpO2 during the 6-minute walking test increased from 86.0 +/- 2.8 (%) to 90.1 +/- 1.3 (%) and dyspnea as measured with Borg scale decreased from 5.6 +/- 1.1 to 4.6 +/- 0.5, although they were not significantly different. These results suggest that pulmonary rehabilitation might improve exercise tolerance in elderly patients with COPD.

Aged↗

[Determination of sucralose in foods by HPLC].

A method for the determination of sucralose in various foods by RI-HPLC and ion chromatography with a pulsed amperometric detector (PAD-IC) was developed. Chopped or homogenized samples were packed into cellulose tubing with 0.01 mol/L hydrochloric acid containing 10% sodium chloride and were dialyzed against 0.01 mol/L hydrochloric acid for 24 hours. The dialyzate was passed through a Bond Elut ENV cartridge, and the cartridge was washed with 0.2 mol/L NaOH and water. Sucralose was eluted from the cartridge with methanol. The extract was taken to dryness in an evaporator and the residue was re-dissolved in water. Sucralose was separated on an Inertsil ODS-3V column with a mobile phase of acetonitrile-water (15:85) and an RI detector. It was also determined on a CarboPak PA1 column with a mobile phase of 100 mmol/L NaOH-75 mmol/L CH3COONa, using a PAD detector. The recoveries of sucralose from various kinds of foods spiked at 50 micrograms/g and 200 micrograms/g ranged from 88-105%. The detection limit in samples was 10 micrograms/g for RI-HPLC and 1 microgram/g for PAD-IC.

Chromatography, High Pressure Liquid↗