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This study aimed to construct a measurement system with the same performance as a measurement system using an automated analyzer and immunoturbidimetric reagents (comparative method) using a flow-type immunosensor (FIS) based on the fluorescence-linked immunosorbent assay technology. In the FIS constructed in this study, all control samples were within the indicated values. The coefficient of variation of repeatability and intermediate precision were less than 2.4% and less than 4.4%, respectively. The lower limit of quantification in this measurement system was 3.9 mg/L, and linearity was confirmed for quantification values, ranging from 3.9 to 465 mg/L. Canine plasma samples (N = 39) were used to measure C-reactive protein (CRP) levels using the comparative method (x) and FIS (y). The regression equation between the measurements was y = 1.035 × - 0.002, with a correlation coefficient of 0.9809, indicating a significantly high correlation. Although the Brandt-Altman analysis suggested the possibility of a proportional systematic error between the two measurements, 38 of the 39 canine plasma samples measured fell within the acceptable range of error, indicating that the measurements are highly consistent. These results suggest that the analytical accuracy of the FIS constructed in this study and the quantitative value of canine CRP are equivalent to those of measurement systems using automated analyzers and immunoturbidimetric reagents.

Lactate dehydrogenase (LDH) in blood is measured using the Japanese Society of Clinical Chemistry (JSCC) method in Japan and the International Federation of Clinical Chemistry (IFCC) method in other countries. In human clinical practice, the IFCC method replaced the JSCC method due to international standardization. Moreover, veterinary LDH measurement will also eventually shift to the IFCC method. However, the relationship between the IFCC and JSCC methods for LDH in various animals and whether they can be equated or not have not yet been investigated. This study aimed to present the changes in measurements in canines and felines after switching to the IFCC method. The plasma LDH activity of canines (N=177) and felines (N=115), who visited a secondary care veterinary clinic, was measured using the JSCC and IFCC methods. The IFCC/JSCC ratio was1.0 in 85% of canines and 88% of felines, indicating that the IFCC method tended to show lower values than the JSCC method, presumably because LDH5 is dominant among the LDH isozymes in canines and felines. The increase in the systematic errors of both assays was in the high value range, with some samples exceeding the error tolerance from near the upper end of the reference range. When switching to the IFCC method for LDH measurement in canines and felines, each institution should consider whether the reference range and clinical diagnostic values established by the JSCC method are appropriate for continued use.

There has been an increase in temperature and the incidence of extreme weather events, such as heat wave, due to global warming, which has promoted the incidence of livestock diseases. Therefore, it is important to examine the effect of changes in environmental parameters on livestock performance. The aim of this study was to examine the relationship between ambient environmental conditions in livestock pen and the physiological parameters of Holstein dairy cows. The results showed that there was a decrease in the red blood cell counts, hemoglobin concentrations, and mean corpuscular hemoglobin concentration of the cows with increasing pen temperature, wet bulb globe temperature (WBGT), and temperature humidity index (THI). Additionally, high daily variation in temperature caused a decrease in the serum albumin levels of the cows. Moreover, the lowest serum calcium, inorganic phosphorus, and magnesium concentrations were observed in November, and were negatively correlated with the 24-hr temperature, WBGT, and THI range of the pen prior to sampling. Multiple regression analysis showed a positive correlation between serum cortisol concentration and 24-hr WBGT range of the pen prior to samplings and packed cell volume. However, serum cortisol and total protein concentrations were negatively correlated. Overall, the findings of the study suggest that large variation in temperature induced stress in the cows, which could be overcome by increased water consumption and improved protein digestion and absorption by the animals, and the addition of minerals, such as calcium to the diet.

Urinary gamma-glutamyltransferase (u-γGT) concentration (U/L) and excretion (urinary creatinine-corrected u-γGT; u-γGT/u-Cre, U/g creatinine) are useful markers for kidney disease. However, there is limited information available on u-γGT and u-γGT/u-Cre distribution in the elderly Japanese population. In this study, we investigated the distribution of u-γGT and u-γGT/u-Cre in 113 Japanese women aged 40–74 years. The u-γGT was assessed from spot urine samples (collected from 09:00 to 14:00) spectrophotometrically according to the Japan Society of Clinical Chemistry reference measurement procedure using l-γ-glutamyl-3-carboxy-4-nitroanilide as the substrate. The u-Cre was measured enzymatically using creatininase, creatinase, sarcosine oxidase, and peroxidase. None of the participants was diagnosed with any kidney disease. Median u-γGT and u-γGT/u-Cre values (central 95% interval values) were 29.7 (5.3–144.0) U/L and 57.9 (32.9–122.7) U/g creatinine, respectively. The distribution of u-γGT tended to decline with age. There was a statistically significant difference in the u-γGT value between the 40-59- and 60-74-year-old groups. In contrast, there was no significant difference in the u-γGT/u-Cre between each age group. The u-Cre level also declined with age. It is suggested that the decline of u-γGT with aging would be masked by the u-Cre correction., Highlights • Median (central 95% interval values) u-γGT value was 29.7 (5.3–144.0) U/L. • Median (central 95% interval values) u-γGT/u-Cre ratio was 57.9 (32.9–122.7) U/g creatinine. • Distribution of u-γGT tends to decline with age. • u-Cre levels also declined with age. • Decline of u-γGT with aging would be masked by the u-Cre correction.

Seaweeds contain large amounts of organoarsenic compounds, mostly arsenosugars (AsSug) and arsenolipids (AsLipid). AsSug is mainly metabolized into dimethylarsinic acid (DMAV) in humans. However, this metabolic process is not well understood. We investigated the metabolism of an AsSug, 3-[5'-deoxy-5'-(dimethylarsinoyl)-β-ribofuranosyloxy]-2-hydroxypropylene glycol (AsSug328), in the gastrointestinal tract using an in vitro artificial gastrointestinal digestion system. AsSug328 was incubated with gastric juice for 4 h, with bile-pancreatic juice for 0.5 h, and finally with enteric bacteria solution for 24 h. The conversion of arsenic compounds after artificial digestion was analyzed by HPLC-ICP-MS and HPLC-ESI-Q-TOF-MS. Our results show that artificial gastrointestinal digestion converted AsSug328 into thio-AsSug328. However, no formation of DMAV was detected. Under the artificial digestion system, the 5-deoxyribofuranose structure of AsSug was maintained. Therefore, AsSug should be absorbed in the intestinal tract after its sugar moiety is partially decomposed. They are then possibly metabolized to DMAV in the liver and subsequently excreted through urine.

河口付近の汽水域は、潮汐や地形等の外的な要因により淡水と海水のバランスが大きく変動することや、人為的な影響を受けやすいことから、河口域における水質は大きく変動する傾向がある。本研究では、利根川河口域を対象として水圏環境調査を行ってきた。その結果、利根川河口域(利根川河口部から河口堰)では潮汐の影響によってその水質環境が大きく変動していた。塩分の調査結果から、満潮時には河口堰底層まで海水が浸入しており、干潮時には河口部までほぼ淡水で占められていた。栄養塩類の濃度は淡水中に高く、河口堰下流部では豊富な栄養塩類を利用して植物プランクトンの活性が高まることが示唆された。また、腸内細菌科細菌(E細菌)数はNaイオン濃度と負の相関関係が得られた。このことから、E細菌が上流からの淡水中に多く存在しており、富栄養化の指標となる栄養塩類と同様の傾向を示すことがわかった。

Objectives: The measurement of the sum of urinary inorganic arsenic (iAs), monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA) has been used for the biological monitoring of occupational ex...

Objectives The sum of urinary inorganic arsenic (iAs), monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA) concentrations is used for the biological monitoring of occupational iAs exposure. Although DMA is a major metabolite of iAs, it is an inadequate index because high DMA levels are present in urine after seafood consumption. We estimated the urinary iAs+MMA concentration corresponding to iAs exposure. Methods We used data from two arsenic speciation analyses of urine samples from 330 Bangladeshi with oral iAs exposure and 172 Japanese workers without occupational iAs exposure using high-performance liquid chromatography with inductively coupled plasma mass spectrometry. Results iAs, MMA, and DMA, but not arsenobetaine (AsBe), were detected in the urine of the Bangladeshi subjects. The correlation between iAs+MMA+DMA and iAs+MMA was obtained as log (iAs+MMA) = 1.038 log (iAs+MMA+DMA) -0.658. Using the regression formula, the iAs+MMA value was calculated as 2.15 and 7.5 μg As/l, corresponding to 3 and 10 μg As/m(3) of exposures, respectively. In the urine of the Japanese workers, arsenic was mostly excreted as AsBe. We used the 95th percentile of iAs+MMA (12.6 μg As/l) as the background value. The sum of the calculated and background values can be used as a biological indicator of iAs exposure. Conclusion We propose 14.8 and 20.1 μg As/l of urinary iAs+MMA as the biological indicators of 3 and 10 μg As/m(3) iAs exposure, respectively.

Chronic inorganic arsenic (iAs) exposure currently affects tens of millions of people worldwide. To accurately determine the proportion of urinary arsenic metabolites in residents continuously exposed to iAs, we performed arsenic speciation analysis of the urine of these individuals and determined whether a correlation exists between the concentration of iAs in drinking water and the urinary arsenic species content.The subjects were 165 married couples who had lived in the Pabna District in Bangladesh for more than 5 years. Arsenic species were measured using high-performance liquid chromatography and inductively coupled plasma mass spectrometry.The median iAs concentration in drinking water was 55 μgAs/L (range0.5-332 μgAs/L). Speciation analysis revealed the presence of arsenite, arsenate, monomethylarsonic acid (MMA), and dimethylarsinic acid in urine samples with medians (range) of 16.8 (7.7-32.3), 1.8 (0.5-3.3), 13.7 (5.6-25.0), and 88.6 μgAs/L (47.9-153.4 μgAs/L), respectively. No arsenobetaine or arsenocholine was detected. The concentrations of the 4 urinary arsenic species were significantly and linearly related to each other. The urinary concentrations of total arsenic and each species were significantly correlated with the iAs concentration of drinking water.All urinary arsenic species are well correlated with each other and with iAs in drinking water. The most significant linear relationship existed between the iAs concentration in drinking water and urinary iAs + MMA concentration. From these results, combined with the effects of seafood ingestion, the best biomarker of iAs exposure is urinary iAs + MMA concentration.

古武術は古くから日本に伝わる武術であり、古武術の体の動かし方をヒントに介護技術に応用したのが「古武術介護」である。全身を連動させ無理なく効率的に動きを引き出すことができれば、介助する人の肩や腰、膝など体にかかる負担が次第に軽減されてゆく。1)古武術介護を実習で行い、学生にアンケートを行った。また従来の搬送法(以下A 法)と古武術介護を使用した搬送法(以下B 法)を実施し、その後バイタルサイン等の結果を比較した。アンケート結果は9 割が負担(抵抗や痛み)を感じず、救急隊員になった時に使いたいと回答し、8 割が古武術介護の各種手技を習熟でき、普段よりも少ない筋力で行うことができたと回答し、7 割が講義中(約3 時間)に古武術介護の手技を体得できたと回答した。この結果から古武術介護技術を習得することは有用である。さらにシミュレーション教育における「搬送」という授業に対して特に筋力・体力が相対的に弱い女性で抵抗感が少なく、モチベーションが上がることが期待できる。バイタルサイン等の結果に有意差はなく、ストレッチャーの上げ下げ時には古武術介護を使用した搬送法の方が腰への負担が少なかったという協力学生の感想から、B 法は救急活動における搬送時に極めて有用である。

臨床検査技師,臨床工学技士,救急救命士の資格は,養成校を卒業したのち,年に1度実施される国家試験に合格しなければ資格を取得することはできない.本学では専任教員による教育,補講の開講,学内模擬試験の実施,外部講師招聘による特別講義を行うなどの試験対策を行っている.しかし,国家試験合格のための学習は,自主学習によるところが大きい.そこで本研究では,臨床検査技師,臨床工学技士,救急救命士等の国家試験の自主学習をサポートするための環境整備を行うことを目的にInformation Technologyを活用した医療系国家試験対策CAIシステムの構築を行った.本CAIシステムは試験問題等のデータベース部分と問題を出題するアプリケーション部分により構成されており,アプリケーションはオブジェクト指向に基づいて設計・開発を行った.これによりデータの追加や管理などとアプリケーションのアップデートを個別に行え,市販のソフトウェアに比べ再利用性・拡張性の高いCAIシステムの構築を実現できた.

The toxicity and carcinogenicity of arsenic depend on its species. Individuals living in Japan consume much seafood that contains high levels of organoarsenics. Speciation analysis of urinary arsenic is required to clarify the health risks of arsenic intake. There has been no report of urinary arsenic analysis in Japan using high performance liquid chromatography with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). We performed speciation analysis of urinary arsenic for 210 Japanese male subjects without occupational exposure using HPLC-ICP-MS. The median values of urinary arsenics were as follows: sodium arsenite (AsIII), 3.5; sodium arsenate (AsV), 0.1; monomethylarsonic acid (MMA), 3.1; dimethylarsinic acid (DMA), 42.6; arsenobetaine (AsBe), 61.3; arsenocholine, trimethylarsine oxide, and unidentified arsenics (others), 5.2; and total arsenic (total As), 141.3 microgAs/l. The median creatinine-adjusted values were as follows: AsIII, 3.0; AsV, 0.1; MMA, 2.6; DMA, 35.9; AsBe, 52.1; others 3.5; and total As, 114.9 microgAs/g creatinine. Our findings indicate that DMA and AsBe levels in Japan are much higher than those found in Italian and American studies. It appears that the high levels of DMA and AsBe observed in Japan may be due in part to seafood intake. ACGIH and DFG set the BEI and BAT values for occupational arsenic exposure as 35 microgAs/l and 50 microgAs/l, respectively, using the sum of inorganic arsenic (iAs), MMA, and DMA. In the general Japanese population, the sums of these were above 50 microgAs/l in 115 (55%) samples. We therefore recommend excluding DMA concentration in monitoring of iAs exposure.

Allele frequencies of 15 short tandem repeat (STR) loci, D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818 and FGA, were analyzed in 127 unrelated Bangladeshi individuals and 105 unrelated Indonesian individuals using the AmpFLSTR Identifiler kit. All STR loci in Bangladeshis and Indonesians were in the Hardy–Weinberg equilibrium.

Allele frequencies and haplotypes for 10 Y-chromosome STR loci, DYS19, DYS385, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS438 and DY439, were determined in 72 unrelated Bangladeshi males using Y-PLEX™5 and Y-PLEX™6 Amplification Kits. This population demonstrated 71 haplotypes, of which 70 were unique. The haplotype diversity calculated from the 10 Y-STR loci was 0.9996 and the discrimination capacity was 0.9861.

銚子市は、「健康スポーツ文化都市宣言」を掲げ、事業の一つとして市民の健康意識の向上を目的に5年前から「健康まつり」を実施している。本学の臨床検査学コースの教員や学生も第1回から、臨床検査技師教育の一翼を担う方法の一つとして参加してきた。この結果、学生からは、検査を受ける人たちへの応対の難しさや、検査値から得られる情報の重要性を再認識したなどの感想が得られ、一方、参加している市民からのアンケートでは、対応が親切で良かったなどの感想が得られた。この「健康まつり」の体験学習を通して、学生は、コミュニケーション能力や臨床病態に関する知識、そして問題解決能力の必要性を短時間で学んだと思われる。また、参加している市民からの感想を得ることができたことは客観的な評価を受けることで体験学習ばかりでなく教育の一環としてフィードバックすることができ、より充実した健康教育につなげることができたと考えられる。

Allele frequencies for four short tandem repeat (STR) loci vWA, TH01, TPOX and F13A01 were determined in three Asian populations, Japanese, Bangladeshis and Indonesians. No deviations from Hardy–Weinberg equilibrium were observed in the three populations for all loci. The combined power of discrimination (PD) for the four loci was 0.9995 in Japanese, 0.99991 in Bangladeshis and 0.9998 in Indonesians.

The polymorphism of FXIIIA was investigated in 105 Indonesians, 141 Bangladeshis, 186 Tibetans, 101 South African Blacks and 100 South African Whites using isoelectric focusing followed by immunoblotting. These population data conformed to the Hardy-Weinberg equilibrium. A circum-pan-Pacific cline in the FXIII * 1A (or FXIII * 1B) allele frequency appeared to exist among the Mongoloids.

Genetic polymorphism of the C1R subcomponent of human complement component C1 was investigated in 105 Indonesians, 141 Bangladeshis, 186 Tibetans, 101 South Africa blacks, 100 South Africa whites, and 24 Samoans. These population data fitted the Hardy-Weinberg equilibrium. A geographical cline of the C1R*1 frequency was observed in Japanese

We have immunohistochemically examined the distribution of the H antigens of type 1, type 2 and type 3/4 chains of the ABO(H) histo-blood group system in human normal colon and in colon cancer using three monoclonal antibodies specific for each of the H type 1/2, H type 2, and the H type 3/4 chain. We unexpectedly found that mucosa of the normal colon from secretors but not that from nonsecretors expressed only H type 1 and did not express H type 2 or H type 3/4. The H type 1 was expressed in goblet cells. Positive goblet cells expressing H type 1 were decreased in number progressively from the proximal colon to the rectum. In tumors, 4 (57%) of 7 cancer tissues of the proximal colon from secretors expressed no H type 1, whereas all 8 cancer tissues of the distal colon from secretors expressed H type 1. The aberrant expressions of H type 2 and H type 3/4 (47 and 67%, respectively) were found in cancer tissues from both the proximal and the distal colon. Tumors from nonsecretors did not express any H antigens. Our results suggested that the expression of H type 1 in the normal colon and the aberrant expressions of H type 2 and H type 3/4 in colon cancer tissues were regulated by FUT2-encoded Se type alpha(1,2)fucosyltransferase. However, UEA-I-positive substance(s) rather than H type 2 were uniquely expressed throughout the normal colon and in colon cancers from both secretors and nonsecretors.

Myotonic dystrophy protein kinase (DMPK)-binding protein, MKBP, has high homology with a small heat shock protein, HSP27. Western blotting analyses showed that MKBP level in rat heart rapidly increased, with a sharp peak at one week after birth (3-fold the level at the fetus), but that it rapidly decreased (1/10 of peak value at 13 weeks). Human myocardium also showed similar age-dependency. Similar but small increase of HSP27 was observed in the neonatal rat myocardium, but not in constitutive and inducible forms of HSP70. Immunofluorescence analysis localized MKBP at the Z lines and intercalated discs in the rat myocardium. MKBP may protect actin cytoskeleton or other proteins of heart muscle against oxidative stress in the neonate.

To detect early changes in neurons and astrocytes by immunohistochemical methods using antibodies against the neuron-specific enolase (NSE), neurofilament, glial fibrillary acid protein (GFAP), and S-100 protein, a fluid-percussion injury model in cats was chosen, in which a severe grade of injury (3.5-5.5 atm) was produced. Neuropathologic changes were produced through brain deformation by pressure gradients at the time of injury. The neuronal NSE immunoreactivity in the parietal cortex and the brain stem began to decrease at 1 to 2 hours after injury and were reduced markedly or even lost 4 hours after injury. Axons in the cerebral white matter and corpus callosum and in the hemorrhage regions at the brain stem were waved and enlarged

Allele frequency data for 15 short tandem repeat (STR) loci, D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818 and FGA, were determined in Tibetan individuals using the AmpFLSTR Identifiler Kit. The agreement with Hardy-Weinberg expectation was confirmed for all studied loci with the exception of FGA.

Transforming growth factor-β1 (TGFβ1) induces a mesenchyme-like cell shape in some epithelial cell types. To clarify the role of TGFβ1 in the morphological regulation of thyrocytes, we performed collagen gel culture of porcine thyrocytes with serum-free medium. TGFβ1-nontreated cells organized follicles. In contrast, the cells treated with 10 ng/ml TGFβ1 became spindle shaped, i.e. they resembled mesenchymal fibroblasts, and did not form follicles. To characterize the spindle-shaped cells, we examined the fine structures and expression of thyroglobulin (Tg) and cytoskeletal proteins using electron microscopy, immunohistochemistry, and immunoblotting. TGFβ1-nontreated cells had microvilli at the apical side facing follicle lumen and had basal lamina at the basal side in contact with collagen gel. TGFβ1-treated cells showed both microvilli and basal lamina at the basal side. TGFβ1-nontreated cells expressed Tg, whereas TGFβ1-treated cells showed no expression. TGFβ1-nontreated cells barely expressed vimentin, but they expressed enough cytokeratin. TGFβ1-treated cells extensively displayed vimentin along with the change in shape to become spindle-like and retained a decreased expression of cytokeratin. TSH (10 mU/ml) did not essentially influence any TGFβ1 effects on the cells. These results indicate that TGFβ1 induces a mesenchyme-like cell shape accompanied by cytoskeletal molecular change and the loss of both epithelial polarization and a function in thyrocytes, and that it results in inhibiting thyroid folliculogenesis with or without TSH.

An immunocytochemical study was performed to examine the expression of cellular c-myc protein in the heart of 30-, 120- and 180-day-old cardiomyopathic Syrian UM-X7.1 hamsters. The heart of age- and sex-matched BIO-RB hamster was used as normal control. In paraffin sections, an immunostaining for c-myc was markedly increased in cytoplasm of cells from the UM-X7.1 heart as compared with that of the BIO-RB heart which showed a weak staining. However, c-myc was localized in nuclei of cells in frozen sections of the heart. Specific cell types of the heart were differentiated with anti-vimentin, and we found that the increased expression of c-myc was present in nuclei of muscle cells of the UM-X7.1 myocardium. A quantitative study of c-myc-positive nuclei of muscle and nonmuscle cells was carried out by a video micrometer. The mean number of c-myc-positive nuclei of muscle cells was significantly higher in the cardiomyopathic heart than in the control heart from hamsters of all ages studied. These results suggest that the increase of c-myc protein may relate to the pathological state or pathogenesis of the hereditary cardiomyopathy.

The microflora in environmental water consists of a high density and diversity of bacterial species that form the foundation of the water ecosystem. Because the majority of these species cannot be cultured in vitro, a different approach is needed to identify prokaryotes in environmental water. A novel DNA microarray was developed as a simplified detection protocol. Multiple DNA probes were designed against each of the 97,927 sequences in the DNA Data Bank of Japan and mounted on a glass chip in duplicate. Evaluation of the microarray was performed using the DNA extracted from one liter of environmental water samples collected from seven sites in Japan. The extracted DNA was uniformly amplified using whole genome amplification (WGA), labeled with Cy3-conjugated 16S rRNA specific primers and hybridized to the microarray. The microarray successfully identified soil bacteria and environment-specific bacteria clusters. The DNA microarray described herein can be a useful tool in evaluating the diversity of prokaryotes and assessing environmental changes such as global warming.

Seaweed contains large amounts of various arsenic compounds such as arsenosugars (AsSugs), but their relative toxicities have not yet been fully evaluated. A risk evaluation of dietary arsenic would be necessary. After developing an arsenic speciation analysis of wakame seaweed (Undaria pinnatifida), we conducted a wakame ingestion experiment using volunteers. Five volunteers ingested 300 g of commercial wakame after refraining from seafood for 5 days. Arsenic metabolites in the urine were monitored over a 5-day period after ingestion. Total arsenic concentration of the wakame seaweed was 34.3 ± 2.1 mg arsenic/kg (dry weight, n = 3). Two AsSugs, 3-(5'-deoxy-5'-(dimethyl-arsinoyl)-β-ribofuranosyloxy)-propylene glycol (AsSug328) and 3-(5'-deoxy-5'-(dimethyl-arsinoyl)-β- ribofuranosyl-oxy)-2-hydroxypropyl-2,3-dihydroxy-propyl phosphate (AsSug482) were detected, but arsenobetaine, dimethylarsinic acid (DMA), monomethylarsonic acid, and inorganic arsenics (iAs) were not detected. The major peak was AsSug328, which comprised 89% of the total arsenic. Approximately 30% of the total arsenic ingested was excreted in the urine during the 5-day observation. Five arsenic compounds were detected in the urine after ingestion, the major one being DMA, which comprised 58.1 ± 5.0% of the total urinary arsenic excreted over the 5 days. DMA was believed to be metabolized not from iAs but from AsSugs, and its biological half-time was approximately 13 h.

Sodium dodecyl sulfate (SDS), an anionic hydrophobic ligand, is known to alter the mechanical properties of elastic fibers. In order to analyze the mechanism of the alteration, two forms of fibrous elastins, "solid" and "powder" elastins, which consisted of fascicular elastic fibers and single or oligomeric elastic fibers, respectively, were prepared from bovine aorta, and the interactions of SDS with these elastins in the presence and absence of 0.15 M NaCl were studied. The solid elastin was able to retain 1.2- to 1.4-fold larger amounts of SDS than the powder elastin under both conditions, and both elastins retained 1.2-fold or larger amounts of SDS in the presence of NaCl than in its absence. Whereas both elastins released the retained SDS gradually on repeated washing with an SDS-free buffer, the release rates from the solid elastin, especially the rate in the presence of NaCl, were much smaller than those from the powder elastin, and the solid elastin retained approximately 40% of the bound SDS under conditions where the powder elastin lost most of its SDS. The SDS-binding capacities of both elastins were significantly lower than those of soluble kappa-elastin and serum albumin, which bound SDS homogeneously on the polypeptide chains. When the washed SDS-bound solid elastin was incubated with methylene blue and examined under a microscope, most of the methylene blue-SDS complex was located at the interfiber spaces of the elastic fibers. These results suggest that SDS alters the mechanical properties of elastic fibers by binding to the interfiber spaces and surfaces of the fibers rather than by binding to the internal polypeptide chains.

Thyroid follicles in vivo are embedded in extracellular matrix (ECM). The composing epithelial cells are in close contact with ECM at the basal side. To examine cell-to-ECM interactions, we studied adhesion, proliferation and differentiation of porcine follicle cells monolayer-cultured on type I and IV collagen, fibronectin or laminin. At 3 h in culture, laminin had the lowest rate of cell adhesion. In proliferation, type IV collagen induced the highest level of nuclear bromodeoxyuridine intake. In a functional differentiation, laminin had about 3 times as much triiodothyronine production as the other ECM molecules. In confluent, culture cells, we also examined an expression of c-fos protein, a transcription factor that plays crucial roles in signal transduction. Immunocytochemistry detected the protein mainly in the nuclei. Western blot showed that laminin induced the highest level of its expression. Thyrotropin (TSH, 10 mU/ml) did not affect adhesion of the cells on any of the substrata or proliferation of the cells on fibronectin ; nor did TSH affect c-fos protein expression of the cells on the substrata except for fibronectin. Our results suggest that type IV collagen and laminin, major components of basement membrane, play positive roles in proliferation and differentiation of follicle cells, respectively, while laminin has no positive effect on adhesion of the cells at early culture ; that the cells express c-fos protein even in contact inhibition of growth and its expression is regulated in part by ECM ; and that ECM controls some behaviors of the cells in a TSH-dependent or TSH-independent manner.

The molecular states of collagen in the aortas of age-matched stroke-prone spontaneously hypertensive (SHRSP) and normotensive Wistar Kyoto rats (WKY) were studied by analyzing its extractability under defined conditions. The monomeric and oligomeric collagen extractable with 0.5 M acetic acid/6 M urea from aortic homogenates of 9-month-old SHRSP and WKY comprised approx. 0.6 and 2.0%, respectively, of the total collagen. On incubation of the acetic acid/urea-extracted residues with pepsin at 4 degrees C, the levels of the collagen alpha 1(I) and alpha 2(I) chains solubilized from the SHRSP residues were both less than 50% of those from the WKY residues. When the residues were incubated with pepsin at 15 or 25 degrees C, the differences became smaller. When the acetic acid/urea residues were hydrolyzed with cyanogen bromide, nearly identical peptide maps were obtained for SHRSP and WKY. The aortas from 2-month-old SHRSP and WKY contained much larger proportions of acid/urea-extractable collagen than those of the older rats (8.2 and 13% of the respective total collagen). The levels of the alpha 1(I) and alpha 2(I) chains solubilizable from the respective residues by pepsin at 4 degrees C were similar to each other. These results indicate that aortic collagen fibrils in SHRSP are stiffened more prominently than those in WKY.

The allele distributions of the STR locus D20S161 have been investigated in Japanese and Bangladeshi people. No deviation from the Hardy-Weinberg equilibrium was observed in either population. D20S161 was found to be a polymorphic STR with seven alleles in Japanese and with six alleles in Bangladeshi. The distribution of allele frequencies in Japanese was very similar to that in Bangladeshi. The D20S161 system can be a potential marker for medicolegal individualization and paternity testing.

論文(Article), Adipose tissue that consists of mature and immature adipocytes is suggested to contain mesenchymal stem cells (MSCs), but a culture system for analyzing their cell types within the tissue has not been established. Here we show that three-dimensional collagen gel culture of rat sc adipose tissue fragments maintained viable mature adipocytes for a long term, producing immature adipocytes and MSC-like cells from the fragments, using immunohistochemistry, ELISA, and real time RT-PCR. Bromodeoxyuridine uptake of mature adipocytes was detected. Adiponectin and leptin, and adipocyte-specific genes of adiponectin, leptin, and PPAR-gamma were detected in culture assembly, whereas the lipogenesis factor insulin (20 mU/ml) and inflammation-related agent TNF-alpha (2 nm) increased and decreased, respectively, all of their displays. Both spindle-shaped cell types with oil red O-positive lipid droplets and those with expression of MSC markers (CD105 and CD44) developed around the fragments. The data indicate that adipose tissue-organotypic culture retains unilocular structure, proliferative ability, and some functions of mature adipocytes, generating both immature adipocytes and CD105+/CD44+ MSC-like cells. This suggests that our method will open up a new way for studying both multiple cell types within adipose tissue and the cell-based mechanisms of obesity and metabolic syndrome.

POPULATION: Seventy-three unrelated Africans living in Cape Town, south region of South Africa.

It has been proposed that in autosomal recessive juvenile parkinsonism (AR-JP), a ubiquitin ligase (E3) Parkin, which is involved in endoplasmic reticulum-associated degradation (ERAD), lacks E3 activity. The resulting accumulation of Parkin-associated endothelin receptor-like receptor (Pael-R), a substrate of Parkin, leads to endoplasmic reticulum stress, causing neuronal death. We previously reported that human E3 HRD1 in the endoplasmic reticulum protects against endoplasmic reticulum stress-induced apoptosis. This study shows that HRD1 was expressed in substantia nigra pars compacta (SNC) dopaminergic neurons and interacted with Pael-R through the HRD1 proline-rich region, promoting the ubiquitylation and degradation of Pael-R. Furthermore, the disruption of endogenous HRD1 by small interfering RNA (siRNA) induced Pael-R accumulation and caspase-3 activation. We also found that ATF6 overexpression, which induced HRD1, accelerated and caused Pael-R degradation; the suppression of HRD1 expression by siRNA partially prevents this degradation. These results suggest that in addition to Parkin, HRD1 is also involved in the degradation of Pael-R.

Allele frequencies of 15 short tandem repeat (STR) loci, D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818 and FGA, were determined for 98 unrelated Africans from South Africa and 98 unrelated Europeans from South Africa using the AmpFlSTR Identifiler PCR amplification kit. The genotype frequency distributions of the 15 STR loci were in the Hardy-Weinberg equilibrium for both populations.

We report here an autopsy case of a 49-year-old woman with depression who died of hyperthermia, probably due to amitriptyline intoxication. She was found dead in bed with several empty amitriptyline pill containers. Her rectal temperature was 41.5°C approximately 3 hours after death. Plasma levels of amitriptyline and nortriptyline were 0.51 and 0.74 mg/l, respectively. Possible mechanisms of fatal hyperthermia are discussed.

The polymorphism of HF (beta 1H-globulin) was investigated in three Asian populations (Bangladeshis, Tibetans and Indonesians) by means of isoelectric focusing and immunoblotting. Phenotypes associated with three common alleles (HF*A, HF*B and HF*Q0) and a rare allele HF*A1 were identified. The observed numbers of phenotypes were in accordance with the numbers expected under the Hardy-Weinberg equilibrium. HF*A1 seems to be a unique allele of the East-Asian Mongoloids including Tibetans and Indonesians.

We used three anti-H monoclonal antibodies (MAbs) specific for H Type 1, H Type 2, and H Type 3/4 antigens to investigate the distribution of H Type 1–H Type 4 chains of the ABO(H) histo-blood group in the human respiratory system. Strong staining of H Type 1 chain and weak staining of H Type 2 chain were observed in mucous cells of submucosal glands of bronchial epithelium, which were dependent on the secretor status. No H Type 3/4 chains were detected in mucous cells. Serous cells of submucosal glands of respiratory system showed no staining by three anti-H antibodies. H Type 1 and H Type 3/4 antigens were detected heterogeneously in apical surfaces of bronchial epithelium from secretors but not from nonsecretors. In contrast, basal cells of bronchial epithelium expressed H Type 2 irrespective of the secretor status, probably regulated by the H gene. Some alveolar Type II cells contained only H Types 3/4, which were dependent on the secretor status, whereas alveolar Type I cells had no H antigens. Our results indicated that different cell types in respiratory epithelium expressed different types of carbohydrate chains of histo-blood group antigens under the control of the H or the Se gene.

The human secretor α(1,2) fucosyltransferase encoded by the FUT2 determines the production of ABO(H) antigens in secretions. Recent studies demonstrated the presence of several nonfunctional alleles in the FUT2. During the analysis for inactivating mutations at the FUT2 locus from 24 Samoan and 47 Bangladeshi individuals, we found two distinct Alu-mediated deletions of FUT2. The FUT2 deletion in a Bangladeshi population was identical with that found in Indian individuals with the Bombay phenotype (sedel), but not associated with the null allele (T725G) of the H gene (FUT1). The FUT2 deletion in Samoans is a novel null allele (sedel2). The junction region of sedel2 was successfully amplified using the same primers for the sedel amplification. DNA sequencing of the junction region of the sedel2 indicated that there was a 32-bp sequence identity between DNA sequences surrounding the 5′ and 3′ breakpoints. The size of the deletion of the sedel2 was 9.3 kb, including the full coding region of FUT2. The frequency of the sedel in a Bangladeshi population was 0.074, and that of the sedel2 in a Samoan population was 0.104. Hum Mutat 16:274, 2000. © 2000 Wiley-Liss, Inc.

The localization and biological roles of the multifunctional cell type mast cells remain unclear in subacute thyroiditis that is characterized by both epithelioid granuloma formation and thyroid tissue repair. We examined their immunolocalization with tryptase of a mast cell marker, using the biopsy specimens from 12 cases. In the epithelioid granuloma, no mast cells were detected in any of the cases, although a small number of them (4.6 +/- 2.4) were seen at the fibrous stroma around the granuloma in all cases. By contrast, in all cases, increased mast cells (28 +/- 7.2) localized at the thyroid tissue-regenerative site where both thyroid folliculogenesis and angiogenesis take place. To elucidate possible roles of mast cells in the disease, we also examined their immunoexpressions of vascular endothelial cell growth factor (VEGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor-BB (PDGF), transforming growth factor-beta1 (TGF-beta1) and epidermal growth factor (EGF), which affect thyroid folliculogenesis and angiogenesis. In all 12 cases, mast cells displayed all of these growth factors in a manner not specific to the infiltrating site. The data suggest that growth factor-expressing mast cells may play crucial roles in the thyroid tissue repair of subacute thyroiditis, modulating thyroid folliculogenesis and angiogenesis; and that the multifunctionality of the cells may be partly dependent on their expressions of various growth factors.

SUMMARY We have investigated by immunochemistry the distribution of H Type 3/4 chains of the ABO histo-blood group system in human submandibular gland using a monoclonal anti-H MBr1 antibody specific for H Type 3/4 chains, and have found the expression of H Type 3/4 chains was mainly in the serous cells. Serous cells from secretors were stained by MBr1 but not by anti-A and anti-B antibodies, whereas serous cells from nonsecretors exhibited a negative reaction with MBr1. Mucous cells were not stained by MBr1. Only a few striated duct cells showed a weak reaction with anti-H MBr1. These results suggested that the H Type 3/4 chains were distributed predominantly in the serous cells of the human submandibular gland and that secretor Type a (1,2)fucosyltransferase (Se enzyme) controlled the synthesis of H Type 3/4 chains in vivo. Saliva also contained H Type 3/4 chains, which were controlled by the secretor gene ( FUT2 ). The differences in the distributions of H Type 1, H Type 2, and H Type 3/4 chains of the ABO histo blood group system in the submandibular gland are discussed. (J Histochem Cytochem 47:889‐894, 1999)

We have examined the immunohistochemical distribution of H Type 1 and of H Type 2 substances of the ABO blood group system in human submandibular gland using either of the two anti-H monoclonal antibodies MAb 1E3 and MAb 3A5. MAb 3A5 was specific for H Type 2, and MAb 1E3 reacted with each of H Type 1-H Type 4 artificial antigens. We have developed a competitive inhibition method against H Type 2 and have obtained MAb 1E3, which is fairly specific for H Type 1 under certain conditions. Mucous cells from secretors were strongly stained by 1E3 and weakly by 3A5, whereas those from nonsecretors showed no reaction with 1E3 and 3A5. Serous cells from both secretors and nonsecretors were stained neither by 1E3 nor by 3A5. Striated and interlobular duct cells were strongly stained by 1E3 and by 3A5, regardless of the secretor status. These results indicated that the expressions of the H Type 1 and H Type 2 in different cell types of the submandibular gland were controlled by different genes. In addition, we have determined the acceptor specificity of two alpha(1,2)fucosyltransferases (H and Se enzymes) after transient expressions of the FUT1 and FUT2 in COS7 cells, and found that the H enzyme activity was similar for both Type 1 and Type 2 precursors, and that Se enzyme activity with the Type 1 precursor was higher than that with the Type 2 precursor. Expression of the H Type 1 antigen in mucous cells was found to be dependent on the Se gene, whereas expressions of the H Type 1 and H Type 2 antigens in striated and interlobular duct cells were dependent on the H gene. (J Histochem Cytochem 46:69-76, 1998)