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Biomedical subjects

Y Furuya

Publications and source records attributed to Y Furuya.

At least 19 recordsLinked to original sources

Nocturnal 5-fluorouracil infusion to patients with breast cancer prior to surgery: appearance of 5-fluorouracil-induced AgNORs aggregation (FAA).

Between 1994 and 1995, 1 day nocturnal infusion of 5-fluorouracil (5-FU) was performed prior to surgery in 13 primary breast cancer patients; 300 mg/m2 of 5-FU was infused constantly from 2100 h to 0700 h via peripheral vein with a volumetric pump. 5-FU concentration in tissues was measured within surgical specimens by HPLC. The concentrations of 5-FU in tumor tissues ranged from 6 to 49 ng/g (average +/- SEM 25.0 +/- 4.1 ng/g), while in normal breast tissues and adipose tissues 5-FU was below the detection limit (<3 ng/g). The 5-FU concentration was lower in estrogen-receptor-positive tumors (14.4 +/- 4.5 ng/g) than in estrogen-receptor-negative tumors (31.8 +/- 5.0 ng/g). Typical FAA was observed in the tumor tissues of three patients. In these three cases, AgNORs were aggregated to one large spheroidal figure in more than 39% of tumor cells. Appearance of FAA could not be predicted by other clinical features. Nocturnal 5-FU infusion caused FAA changes in certain types of primary breast cancer.

Aged

Ultra-high-resolution scanning electron microscopy of the sarcoplasmic reticulum of the rat atrial myocardial cells.

BACKGROUND: The sarcoplasmic reticulum (SR) of mammalian ventricular and atrial muscles share common features but also differ because T-tubules are rare and extended junctional SR is exclusively seen in the atrium. This scanning electron microscope (SEM) study was undertaken to clarify the three-dimensional organization of the rat atrial SR system. Specific preparations were examined with an ultra-high-resolution SEM. METHODS: Fixed right rat atria were frozen, fractured, and macerated by the aldehyde-osmium-DMSO-osmium method to remove myofibrils and cytoplasmic matrix. Left exposed were mitochondria, SR, and sarcolemma. Dried specimens were then impregnated by osmium-hydrazine and examined without metal coating. RESULTS: In place of conventional T-tubules, a prominent type of sarcotubules, Z-tubules, were found at the Z-line level. Branches from these tubules joined the cisternal SR, which was 100-300 nm in diameter and localized near the Z-line, and formed extensive SR meshworks and polygonal patches. Bulbous swellings, the corbular SR, were also evident. Sarcotubular reticulum completely surrounded each myofibril. The intermyofibrillar SR, especially Z-tubules, joined the peripheral subsarcolemmal SR, which was also arranged as a meshwork and was closely apposed to the sarcolemma. CONCLUSIONS: These SEM observations confirm the organization of the rat atrial SR system and present new, detailed, three-dimensional images of Z-tubules, cisternal SR, extended junctional SR, and peripheral SR, which provide further structural insight.

Animals

Competitive NMDA and strychnine-insensitive glycine-site antagonists disrupt prepulse inhibition.

Prepulse inhibition (PPI) is thought to reflect the operation of a sensorimotor gating system in the brain. Sensorimotor gating abnormalities have been identified in schizophrenic patients, and various neural systems are involved in this function. To study the modulation of the sensorimotor gating system by the N-methyl-D-aspartate (NMDA) receptor channel complex, the effects of noncompetitive and competitive NMDA antagonists on PPI were examined in rats. PPI was not disrupted by CGS 19755, a competitive NMDA antagonist, at 30 min after subcutaneous (s.c.) administration. However, CGS 19755 (40 mg/kg s.c.) decreased PPI at 120 min after administration with a marked decrease of startle amplitude. Late onset of the effect of CGS 19755 was also observed in the increase of spontaneous locomotor activity (SLA). On the other hand, phencyclidine, a noncompetitive NMDA antagonist, disrupted PPI at 30 min after administration and increased SLA from 20 min after administration. PPI was also disrupted by bilateral intracerebroventricular administration of 5,7-dichlorokyn urenate (10 and 20 micrograms/side X 2), an antagonist at the strychnine-insensitive glycine receptor, which is an allosteric binding site in the NMDA receptor-channel complex. It is concluded that the NMDA receptor-channel complex plays an important role in regulation of PPI.

Acoustic Stimulation

Synthesis of L-2,4-diamino[4-11C]butyric acid and its use in some in vitro and in vivo tumour models.

L-2,4-Diamino[4-11C]butyric acid (DAB) was synthesized by an enzyme catalysed carrier added (0.1 micromol KCN) reaction of hydrogen [11C]cyanide with O-acetyl-L-serine followed by reduction. L-[11C]DAB was obtained with a radiochemical purity higher than 96% and with a decay corrected radiochemical yield of 30-40% within a 32 min reaction time. The enantiomeric excess was 98%. The uptake of L-[11C]DAB was investigated in multicellular aggregates of six different cell lines and animal tumour models. L-[11C]DAB is potentially useful for the assessment of pharmacokinetics of L-DAB in vivo for part of its evaluation as an antitumoural agent, although its use for diagnostic purposes seems limited.

Aminobutyrates

Occurrence of high ratio of males after introduction of minocycline in a colony of Leptotrombidium fletcheri infected with Orientia tsutsugamushi.

In colonies of Leptotrombidium fletcheri mites infected with Orientia tsutsugamushi (Ot), the agent of scrub typhus, males rarely appear. In the present study, the development of a high ratio of males was observed after introduction of minocycline (MC). A high dose of MC was injected subcutaneously into a mouse, and by feeding unfed larvae from an infected mite colony on this mouse, the Ot in the mites were successfully killed. Of a total of 130 unfed larvae attached to the mouse, 29 developed into females; of these, 9 laid an average of 112.4 eggs/female. Unfed larvae in the succeeding generations were attached to untreated mice. All adults in the P and F1 generation were females, and males started to appear at the F2 generation. The ratio of males to females was 332:7, 108:13, 263:61 and 71:9 at the F2, F3, F4 and F5 generations, respectively. These data suggest that Ot in the ovary or gonad may suppress the development of males.

Animals

Lectin histochemistry in rat liver fibrosis induced by heterologous serum sensitization.

The localization of carbohydrates in rat livers with fibrosis induced by heterologous serum was examined by lectin histochemical and biochemical techniques. Twenty-four lectins were used to visualize the different carbohydrates in paraffin sections of normal and fibrotic liver tissues. No differences in staining patterns of these lectins were observed between normal and fibrotic livers in hepatocyte cell membranes including bile canaliculi, sinusoidal endothelial, or bile ductal cells. Kupffer cells strongly stained with Vicia villosa agglutinin (VVA) were seen only in the periportal zone of the normal liver, but they were observed in the periportal zone and scattered throughout the pseudolobular zone in the fibrotic liver. The cytoplasm of some hepatocytes was strongly stained by Bandeiraea simplicifolia lectin-I (BSL-I). BSL-I positive hepatocytes in normal liver were localized in the periportal zone, but those in the fibrotic liver were scattered in the periportal and perifibrous zones. After polyacrylamide gel electrophoresis of liver glycoproteins, differences in molecular sizes of BSL-I positive glycoproteins (79 and 81 kD) were detected by lectin blotting. Cell density of perifibrous BSL-I positive hepatocytes may be useful as a diagnostic parameter for liver fibrosis and/or cirrhosis. Two distinct staining patterns with twelve lectins were observed in fibrotic septa of the fibrotic liver. The fibrotic septa were stained with six characteristic lectins, and the centrilobular septa were stained with all these twelve of lectins. Histopathological assessment of the centrilobular fibrotic septa stained with these characteristic lectins may contribute to the diagnosis and prognosis of hepatic fibrosis.

Animals

[Mechanism on androgen-independent progression of prostate cancer].

Eighty percent of prostate cancer with metastasis respond to androgen ablasion, showing initial androgen-sensitive growth. However, more than half of responders gradually loses dependency up to 5 years. Animal experiments reveal that loss of androgen sensitivity is attributable to complex reasons; adaptation, paracrine control by other androgen-independent tissues, genetic changes and mutation of androgen receptor. Most important event is explained from alteration of expression on oncogenes and suppressor genes. Counterplan of the progression was discussed.

Androgen Antagonists

Fluorescence photography as a diagnostic method for oral cancer.

This study was carried out to evaluate the diagnostic utility of autofluorescence photography for oral mucosal lesions. The materials consisted of 15 chemically-induced lesions containing carcinomas in 15 hamsters, and 32 oral lesions in 30 patients. In the animal models, orange fluorescence was detected in all squamous cell carcinomas invading the muscle layer, and the intensity of the fluorescence increased with the progress of the lesions. In the clinical application, orange fluorescence was detected in 14 of 16 malignant tumors and in one of 16 benign lesions. These results suggest that fluorescence photography may be useful for the diagnosis of oral cancer, particularly for squamous cell carcinoma.

9,10-Dimethyl-1,2-benzanthracene

Identification of histone H2A.X as a growth factor secreted by an androgen-independent subline of mouse mammary carcinoma cells.

Shionogi carcinoma 115 (SC 115) cells and Chiba subline 2 (CS 2) cells are clones of an androgen-responsive mouse tumor cell line and its autonomous subline, respectively. We have shown previously that CS 2 cells produce a heparin-binding growth factor that stimulates the growth of SC 115 cells as well as the growth of themselves. In this study, a growth factor was purified from serum-free conditioned media of CS 2 cells cultured without testosterone. A heparin-binding fraction showed growth- promoting activity on SC 115 cells and BALB/3T3 cells. The amino acid sequence analysis revealed that the components were identical to histones H2A.1 and H2A.X. Since histone H2A purified from bovine thymus had almost no growth-promoting activity on SC115 cells, histone H2A.X was assumed to be a growth factor. cDNA of histone H2A.X was cloned from a library of CS 2 cells, and its sequence was confirmed. The expressed product of histone H2A.X cDNA in Escherichia coli showed remarkable stimulatory effects on growth of SC 115 cells cultured in the absence of testosterone. These results indicate that histone H2A.X is secreted from CS 2 cells cultured without testosterone and plays a role as a growth factor.

3T3 Cells

Induction of apoptosis in androgen-independent mouse mammary cell line by 1, 25-dihydroxyvitamin D3.

Androgen-dependent tumors eventually progress to independent tumors after androgen withdrawal. Effective treatment for hormone-independent tumors is therefore needed. Androgen-independent CS-2 cells could grow in serum-free culture whether androgen is present in the medium or not. In the present study, the mechanism of cell death in CS-2 cells was examined after 1, 25-dihydroxyvitamin D3[1, 25(OH)2D3] treatment. 1, 25(OH)2D3 has been examined as an anti-tumor agent, but its role in promoting cell death is poorly understood. Based upon the temporal sequence of DNA fragmentation, morphologic changes and loss of cell viability, the cells underwent apoptosis with 1, 25(OH)2D3 treatment. Northern-blot analysis was used to identify a series of genes whose expression per cell is enhanced during the apoptotic pathway. In the apoptotic process induced by 1, 25(OH)2D3, mRNA expression of testosterone-repressed prostatic message 2, transforming growth factor beta1, glucose-regulated 78-kDa protein and calmodulin increased. Flow-cytometric analysis showed that 1, 25(OH)2D3 treatment resulted in a block in G0/G1 of the cell cycle. These results demonstrate that androgen-independent CS-2 cells retain the ability to undergo apoptosis by 1, 25(OH)2D3. This system appears to be a good model for investigating apoptosis of hormone-independent cancer.

Actins

Dopamine D3 agonists disrupt social behavior in rats.

Behavioral studies were conducted in rats administered a selective D3 agonist, 7-hydroxy-N,N-di-n-propyl-2-aminotetralin (7-OH-DPAT) or 4aR, 10bR-(+)-trans-3,4,4a,10b-tetrahydro-4-propyl-2H,5H-[1] benzopyrano[4,3-b]-1,4-oxazin-9-ol (PD 128907). Both drugs induced disruption of huddling behavior in rats at doses that did not produce hyperlocomotion. The effects of the D3 agonists were dependent upon dosage and time after administration. These results suggest that D3 receptors are concerned with social interaction in rats.

Animals

Title aggregation patterns of argyrophilic nucleolar organizer regions induced by 5-fluorouracil in the nuclei of MCF-7 human breast cancer cells.

The effects of tamoxifen and 5-fluorouracil (5-FU) on the patterns of argyrophilic nucleolar organizer regions (AgNORs) in MCF7 human breast cancer cells were studied. Tamoxifen and 5-FU both inhibited the growth of MCF-7 cells by 18% by day 3 of culture, but each had different effects on the AgNORs. Whereas no significant changes were induced by tamoxifen, effects on the AgNORs of MCF-7 cells by 5-FU were dramatic: 5-FU treatment changed the pattern of AgNORs, reducing the number of satellites by aggregation, typically to a single aggregation around nucleoli in a sphenoidal fashion. We named these morphological changes: fluorouracil induced AgNOR aggregations (FAA). Following treatment with 500 ng/ml 5-FU, FAA developed rapidly. AgNORs forming two or three aggregates in 24% (6 h), 24% (12 h), 40% (24 h) and 34% (48 h) of cells, compared to a control rate of 14%. Single large aggregate was rarely found in untreated cultures but after 6, 12, 24 and 48 h treatment with 500 ng/ml 5-FU, AgNORs had formed a single aggregate in 6, 8, 16 and 22% of cells, respectively. FAA were observed at a concentration of 100 ng/ml 5-FU; 48 h treatment resulted in cells in which two or three aggregates were increased by 24% and single aggregate by 16%. These large single aggregates were larger than nucleoli stained by Papanicolau staining.

Antimetabolites, Antineoplastic

Identification of the five essential histidine residues for peptidylglycine monooxygenase.

Peptidylglycine monooxygenase (PGM) is a copper-containing monooxygenase that plays a key role in the peptide C-terminal alpha-amidation. Comparative analysis of the amino acid sequences of rat, human, bovine and frog PGMs revealed that ten histidines (residues 107, 108, 172, 235, 242, 244, 279, 364, 366 and 367 in rat PGM) are conserved among the four species. We introduced site-directed mutations to the ten histidines of rat PGM and found that the mutation of His- 107-->Ala, His-108-->Ala, His-172-->Ala, His-242-->Arg or His-244-->Ala abolished the enzyme activity. The five mutant proteins lacking the enzyme activity bound to a substrate, Phe-Gly-Phe-Gly, as did the wild type PGM. These results along with available evidence indicate that the five histidine residues (His-107, 108, 172, 242 and 244) are essential for PGM activity, acting as copper ligands.

Amino Acid Sequence

Comparison of mucosal microvasculature between the proximal and distal human colon.

The microcirculation of the human colon with special reference to the differences in microvascular architecture between the proximal and distal colon was studied by scanning electron microscopy with vascular corrosion casting technique. The subsurface capillary networks of the ascending colon were honeycomb-like and multi-layered, with an average number of capillary layers per capillary loop of 3.28 +/- 1.10 (mean +/- SD). Whereas, those of the sigmoid colon were almost single-layered, and the average number of capillary layers was 1.19 +/- 0.39. In the cross-section of vascular casts of both parts of the colon, the ascending capillaries originating from the submucosal arterioles ascended into the mucosal layer and joined into the subsurface capillary networks, which drained into the collecting venules near the surface. The mean diameter of the collecting venules of the ascending and the sigmoid colon was 29.3 +/- 6.41 microns and 19.48 +/- 2.23 microns, respectively. From these findings, it is speculated that the multi-layered capillary networks of the proximal colon are closely related to the greater water absorption and electrolyte transport activities compared to those of the distal colon.

Aged

[Spotted fever group rickettsiosis and vectors in Kanagawa prefecture].

Primer pairs for PCR were designed from the gene encoding the 17,000-molecular-weight genus-common antigen of Rickettsia japonica, Rickettsia rickettsii, Rickettsia conorii, Rickettsia typhi and Rickettsia prowazekii. Primers R1, R2 were designed for amplifying the genomic DNA from spotted fever group (SFG) rickettsiae and epidemic typhus rickettsiae. Primers Rj5, Rj10 were designed for amplifying the genomic DNA from only R. japonica. Using the primers R1, R2, about a 540-bp fragment was observed by amplifying the genomic DNA from R. japonica, R. rickettsii, R. conorii, Thai tick typhus TT-118, Rickettsia sibirica, Rickettsia montana, Rickettsia askari, R. typhi, R. prowazekii and Katayama strain isolated from the patient infected with SFG rickettsiae. Using the primers Rj5, Rj10, the 357-bp fragment was observed by amplifying the genomic DNA from R. japonica and Katayama strain. Therefore, the Katayama strain was identified to belong to R. japonica. With primers R1, R2 and Rj5, Rj10, 537 bp and 357 bp bands were amplified from blood of the patients infected with SFG rickettsiae in Kanagawa prefecture. These findings indicate that the causative agent of SFG rickettsiosis in these two patients was R. japonica. The ticks, Ixodes ovatus and Haemaphysalis flava, were collected by out field research in Kanagawa prefecture. With primers R1, R2 and Rj5, Rj10, 537 bp and 357 bp were amplified from these ticks. This indicates that I. ovatus and H. flava were the vector of R. japonica in Kanagawa prefecture. Also, with the primers R1, R2, about a 540 bp fragment was amplified but with primers Rj5, Rj10, no fragments were amplified from I. ovatus and H. flava. Therefore, these ticks may have SFG rickettsiae other than R. japonica and epidemic typhus rickettsiae.

Animals

[Accuracy of a pulse oximeter during hypoxia].

The accuracy of the pulse oximeter was examined in hypoxic patients. We studied 11 cyanotic congenital heart disease patients during surgery, and compared the arterial oxygen saturation determined by both the simultaneous blood gas analysis (CIBA-CORNING 288 BLOOD GAS SYSTEM, SaO2) and by the pulse oximeter (DATEX SATELITE, with finger probe, SpO2). Ninty sets of data on SpO2 and SaO2 were obtained. The bias (SpO2-SaO2) was 1.7 +/- 6.9 (mean +/- SD) %. In cyanotic congenital heart disease patients, SpO2 values were significantly higher than SaO2. Although the reason is unknown, in constantly hypoxic patients, SpO2 values are possibly over-estimated. In particular, pulse oximetry at low levels of saturation (SaO2 below 80%) was not as accurate as at a higher saturation level (SaO2 over 80%). There was a positive correlation between SpO2 and SaO2 (linear regression analysis yields the equation y = 0.68x + 26.0, r = 0.93). In conclusion, the pulse oximeter is useful to monitor oxygen saturation in constantly hypoxic patients, but the values thus obtained should be compared with the values measured directly when hypoxemia is severe.

Adolescent

[Bacterial contamination of anesthesiologists' hands and the efficacy of handwashing].

The purpose of this study was to examine bacterial contamination, especially by transient skin flora, that were on the hands of trainee and diplomate anesthesiologists during general anesthesia as well as to evaluate the efficacy of washing hands with running water or with three alcohol based antiseptic solutions using a modified glove juice method. The bacterial counts on the anesthesiologists' hands were 3.21 +/- 0.66 [log10 (mean +/- SD)] during induction, 255 +/- 1.15 during maintenance of anesthesia, 2.67 +/- 1.10 during extubation and 3.57 +/- 0.74 at the end of anesthesia. The diplomates' hands were more contaminated than those of the trainees during both intubation and extubation. Washing hands with running water or antiseptic solutions was effective to reduce bacterial contamination, but there was no disinfectant effect of antiseptic solutions against the bacteria that adhered to the hands after drying those solutions. Therefore to prevent nosocomial infection, anesthesiologists should wash their hands with running water or the antiseptic solution after each contact.

Anesthesia, General