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Although studies on snow algae and macroinvertebrates have been frequently conducted on snow patches, only few surveys have been focused on microinvertebrates which reach high biomass and play various trophic roles in other cold habitats. The aims of this study were (1) to search for microinvertebrates in seasonal surface snow patches located on the slope of Mt. Gassan, in northern Japan, and (2) to identify factors determining their distribution associated with snow algal blooms of various colorations (orange, green, and golden-brown) collected from the same sampling site over two seasons (2018, 2019). Microscopic observation revealed presence of two major groups of microinvertebrates: Tardigrada and Rotifera. They were concentrated in green snow colored by blooms of Chloromonas sp. in comparison to orange or golden-brown snow and only a few were found in white snow. Mean body length of tardigrades increased throughout the melt season, their intestine content was green and they laid eggs on colored snow. These results suggest that tardigrades preferentially grew and reproduced on green snow patches. Population densities of tardigrades, rotifers and concentration of chlorophyll a were significantly correlated. Our study indicates that green snow patches in temperate mountainous forests constitute important and unique low-temperature ecosystems for microinvertebrates. Snow covered by algae is an unrecognized novel habitats for tardigrades and rotifers.

The results show the morphological analyses and spectroscopic studies of snow and glacier algae and their parasitic fungi in Svalbard (High Arctic). Fixed algal cells of two species, Sanguina nivaloides and Ancylonema nordenskioeldii, were imaged using light microscopy, scanning electron microscopy (SEM), and atomic force microscopy (AFM). Fluorescence microscopy using Calcofluor white stain supported the observations of parasitic fungi on the algal cells. Images in brightfield microscopy showed chytrid-like fungi penetrating the cells of both algal species. Parasites were found to colonize the cells of A. nordenskioeldii and hypnozygotes of S. nivaloides, while no fungi infected the cyst stages of S. nivaloides. The autofluorescence analysis revealed the ability of S. nivaloides to glow when excited with different wavelengths, while A. nordenskioeldii did not fluoresce. The hypnozygotes of S. nivaloides emitted brighter fluorescence than the cysts, and the most intense luminosity was observed in the UV range. The Fourier-transform infrared spectroscopy (FTIR) and energy-dispersive X-ray spectroscopy (EDS) spectroscopic analysis showed differences in the chemical composition between samples collected from three different sites. Samples dominated by cyst cells were characterized by the presence of an abundant polysaccharide envelope.

Pollen grains are commonly found in ice cores, particularly those from mountain glaciers at low to middle latitudes. Because the release of pollen from flowers has a seasonality and varies among the plant species, pollen concentrations in ice cores are useful for distinguishing annual or seasonal layers and inferring past vegetation near glaciers. We analyzed major pollen grains in an 87-m-deep ice core drilled at the top of the Grigoriev Ice Cap (4563 m.a.s.l.) in the Tien Shan Mountains, Kyrgyz Republic. Microscopy showed that mainly five pollen taxa were contained in the ice core. Their abundance fluctuated with the depth of the core, indicating their seasonal deposition on the ice cap, while the seasonality of the stable isotopes was not particularly clear because of melting and refreezing signatures. Based on the pollen profiles, annual layers were determined back in time to 1780 AD at a depth of 63.6 m; at greater depth, they could not be distinguished due to ice layer thinning in the glacier. The pollen assemblages have gradually changed during the last 220 years and were particularly distinctive in the deeper part, suggesting the drastic change of vegetation in this region during the Holocene.

Cryoconite holes are water-filled cylindrical holes formed on ablation ice surfaces and commonly observed on glaciers worldwide. Temporal changes of cryoconite holes characteristically −2, 40–55% of the incoming flux). In contrast, holes developed in sunny conditions correspond to high solar radiation (186–278 W m−2, 63–88%). Results suggest that the dimensions of holes drastically changed depending on the weather conditions and that frequent cloudy, warm and windy conditions would cause a decay of holes and weathering crust, inducing an increase in the cryoconite coverage on the ice, consequently darkening the glacier surface.

Recent molecular data has strongly suggested that field-collected cysts of snow algae that are morphologically identifiable as the zygotes of Chloromonas nivalis are composed of multiple species. Motile vegetative cells, however, have not been directly obtained from these cysts because of the difficulties involved in inducing their germination. Recently, our comparative molecular analyses, using both field-collected and cultured materials, demonstrated that one Japanese lineage of "C. nivalis zygotes" belongs to C. miwae. Herein, we examined another Japanese lineage of field-collected "C. nivalis zygotes" and a new strain originating from Japan. Our molecular data demonstrated that these two different life cycle stages are conspecific, and that they represent a new species that we herein describe as C. muramotoi sp. nov., based on the vegetative and asexual morphological characteristics of the strain. Multigene phylogenetic analyses showed that this new species was sister to C. miwae. Scanning electron microscopy demonstrated that the cysts of C. muramotoi are different from those of C. miwae, based on the arrangement of the flanges developing on the cell wall.

Glacial meltwater samples were collected from 15 supraglacial ponds at elevations ranging between 3989 and 4292 m above sea level in the debris-covered area of Lirung glacier in central Nepal Himalayas, from November 2010 to October 2011 on a bimonthly basis. We examined elevation trends, and their control by seasonality, dominant geochemical processes, and seasonal dynamics in these supraglacial ponds. Concentration of some parameters showed a decreasing trend with elevation but with high variability and modulation by seasons, being more prevalent in the post-monsoon. Sulfate is the only parameter showing significant elevation trends regardless of the season, although the trend is also affected by the monsoon. The concentrations of major chemical solutes were highest during post-monsoon season and lower in the pre-monsoon and monsoon seasons, indicating influence of seasonality on dissolution, dilution, and hydrological processes. Contribution to chemical loads from marine aerosols to these pond water appeared to be minor except for sodium and magnesium. Chemical weathering of carbonate and silicate minerals fueled by dissolution of pyrite appeared as the dominant geochemical processes.

Antibiotic resistance genes are biologically transmitted from microorganism to microorganism in particular micro-environments where dense microbial communities are often exposed to an intensive use of antibiotics, such as intestinal microflora, and the soil microflora of agricultural fields. However, recent studies have detected antibiotic-resistant bacteria and/or antibiotic resistance genes in the natural environment geographically isolated from such areas. Here we sought to examine the prevalence of antibiotic resistance genes in 54 snow and ice samples collected from the Arctic, Antarctic, Central Asia, North and South America and Africa, to evaluate the level of these genes in environments supposedly not affected by anthropogenic factors. We observed a widespread distribution of antibiotic resistance genes in samples from various glaciers in Central Asia, North and South America, Greenland and Africa. In contrast, Antarctic glaciers were virtually free from these genes. Antibiotic resistance genes, of both clinical (i.e. aac(3), blaIMP) and agricultural (i.e. strA and tetW) origin, were detected. Our results show regional geographical distribution of antibiotic resistance genes, with the most plausible modes of transmission through airborne bacteria and migrating birds.

In this study, we used a 4.00-m pit on Belukha glacier in Russia's Altai region and attempted to establish the timing of chemical deposition events by analyzing pollen profiles. As the pollen deposition of each examined taxon on the glacier surfaces followed a distinct seasonal phenology, seasonal layers could be identified over a two-year period. The seasonal layer boundaries reconstructed from the pollen analyses were in close agreement with the in situ observations and indicated that the snow deposition on the glacier originates mainly from summer precipitation. The record of oxygen isotope ratios showed a relatively high mean value of −13.3‰, which was attributed to the absence of winter depositions. The formate (HCOO−) concentration records displayed seasonal variation with the highest emissions occurring in the spring, and a dust event in the spring of 2003 was detected from the Mg2+, Ca2+, and dust concentration profiles. Taken together, these results suggest the analysis of pollen profile...

Reliable chronologies in ice cores and snow pits from many alpine glaciers in latitudes between 60° N and 60° S are often difficult to establish owing to problems with annual-layer counting. Problems arise from melting, wind erosion and the negligible amount of precipitation in some seasons, all of which tend to obscure the seasonal variations in δ18O and chemical concentrations that are typically used to date ice cores. However, alpine glaciers contain many species of pollen grains that peak at particular times of the year. We used the peaks in Betulaceae, Pinus, Artemisia and a combination of Abies and Picea pollen species to determine the four seasonal layers of a snow pit on Belukha glacier in Russia’s Altai Mountains. Comparing the pollen-dated profiles with wind and precipitation records allows us to determine where a seasonal layer is missing. Thus, the pollen-dating method described here may be a useful tool to measure the annual snow deposition on alpine glaciers, even when some seasonal layers are eroded by wind or missing due to negligible precipitation.

We have developed a new ELISA for quantifying N-acetyl-β-d-glucosaminidase (NAG) isoenzyme B in human urine after raising monoclonal antibodies against the isoenzyme from human placenta. Though the obtained antibodies reacted not only to isoenzyme B but also to A, we could detect isoenzyme B selectively by a two-step sandwich ELISA with a pair of selected antibodies at low pH in the first reaction. The detected limit was 0.5 μg/L for a sample volume of 25 μL. Within-run CVs ranged from 2.5% to 5.4% and between-run CVs ranged from 6.2% to 9.1%. Recoveries of NAG isoenzyme B added to each of three urine samples ranged from 91% to 114%. The dilution curves of urine samples showed good linearity. The cross-reactivity of NAG isoenzyme A was practically negligible (2–3%). The mean value for NAG isoenzyme B in spot urines from healthy adults was 2.9 μg/g creatinine. This ELISA method is rapid and precise enough for routine determination of NAG isoenzyme B in human urine.

Heparin administration to diabetic rats caused no change in VLDL, an increase in IDL and a decrease in LDL on electrophoretic analysis of plasma lipoproteins, while the administration to control rats markedly decreased VLDL and increased IDL and LDL. Both hepatic triglyceride lipase (HTGL) and lipoprotein lipase (LPL) activities in the postheparin plasma were lower in the diabetic rats than in the controls, and the reduction of HTGL activity was greater than that of LPL activity in the diabetic rats. The LPL activity in the adipose tissue was lower in the diabetic rats than in the controls, but the activities in the cardiac and skeletal muscles were similar in the two rats. The HTGL-catalyzed fatty acid (FA) releases from the diabetic VLDL and IDL were lower than those from the normal rat VLDL and IDL, while the LPL-catalyzed FA release in the diabetic rats was not different from those in the controls. The decreases in LPL and HTGL activities and the markedly impaired susceptibility of IDL to HTGL coincide well with the postheparin changes in plasma lipoproteins in diabetic rats, an increase in IDL and a decrease in LDL.

Intense optically stimulated luminescence (OSL) with a peak at about 430 nm is observed in ultraviolet (UV)‐ray or x‐ray irradiated europium‐doped potassium chloride (KCl:Eu) phosphor crystals. The OSL intensity is increased with increasing UV‐ray or x‐ray irradiation dose. This suggests that KCl:Eu phosphor crystal is useful as a material for two‐dimensional UV‐ray or x‐ray imaging sensor utilizing OSL phenomenon. The results obtained are consistent with the proposed emission mechanisms of the 420 nm OSL peak, based on the recombination of electrons released from the F centers with Eu3+ ions. The excitation mechanism for the OSL in UV‐ray irradiated KCl:Eu phosphor crystals is also discussed.

Rats with alloxan-induced diabetes developed severe atherosclerotic lesions when they were maintained on a 0.25% cholesterol diet for one year. The atheromatous changes developed at the aortic arch, appeared as early as 3 months after the start of the experiment, and increased thereafter. The diabetic rats also developed atherosclerosis when they were fed standard rat chow, but the area of the atheromatous lesion was about one tenth of that in rats fed the high-cholesterol diet. Normal rats did not develop atherosclerosis even when fed the high-cholesterol diet for one year. The alloxan diabetic rats showed no increase in body weight, but developed serum glucose levels as high as 600-800 mg/dl as well as high serum cholesterol levels and lower serum HDL-cholesterol levels. The development of atherosclerosis in these rats was significantly related to an increase in the serum cholesterol/phospholipid ratio, the atherogenic index (TC-HDLC/HDLC), and the serum total cholesterol level, but was not related to the serum glucose, HDL-cholesterol, triglyceride, or lipid peroxide levels. These relationships were found as early as B-16 weeks after the start of the experiment. These data suggest that the serum cholesterol/phospholipid ratio, the atherogenic index, and the total cholesterol level are important risk factors for the development of atherosclerosis in rats with alloxan diabetes.

In vitro glycation of very low density lipoprotein (VLDL) reduced the susceptibility to lipoprotein lipase (LPL) as the level of glycation increased. Addition of reduced glutathione to an incubation medium of serum and glucose interfered with glycation of serum proteins when the concentration of reduced glutathione was higher than 10 mM. At concentrations higher than 25 mM, it also significantly prevented the glycation induced reduction of fatty acid releases from VLDL by LPL. There were no such effects on glycation of serum protein and the fatty acid release from the addition of aminoguanidine. By contrast, addition of D-lysine enhanced glycation of serum proteins by glucose and further decreased fatty acid release from VLDL by LPL. From these results, it is suggested that glycation of VLDL decreases the susceptibility of VLDL to LPL. Delayed catabolism of VLDL in diabetic patients is considered partly caused by glycation of apoproteins, which renders VLDL less sensitive to LPL, in addition to the decreased LPL activity in diabetes mellitus.

Alloxan-diabetic rats were kept on an ordinary or 0.25% cholesterol supplemented diet for up to 12 months, and changes in the serum lipid and lipoprotein levels were examined. The diabetic rats showed IRI levels of 3-5 IU/ml, which were 1/5-1/10 that of the normal rats, and about 10 times higher serum glucose levels than the normal rats. The diabetic rats consumed 1.5-2 fold amounts of diet but their body weights were almost constant. The diabetic rats showed marked hyperlipidemia, especially in the cholesterol and triglyceride levels. The phospholipid level also increased but for a lesser extent and the c/p ratio increased. The serum lipoprotein levels increased in the diabetic rats, 5-10 times in the ordinary diet fed groups and 10-20 times in the cholesterol diet fed groups. Chylomicron, VLDL, IDL and LDL increased but HDL decreased. These changes were more significant in the diabetic rats fed the cholesterol diet. The triglyceride levels in the lipoprotein fractions increased in the diabetic rats, but the increase in cholesterol was more significant and the relative composition ratios of triglyceride decreased. The composition ratios of phospholipids, however, remained almost constant in all the lipoprotein fractions. The increase in IDL, especially cholesterol in the IDL fraction, was remarkable in the diabetic rats. The responsiveness of the diabetic rat VLDL to lipoprotein lipase (release of free fatty acids) was less than a half of that of the normal rat VLDL. In conclusion, the diabetic rats showed an increase in triglyceride but more significant increase in cholesterol in the lipoprotein fractions. These changes in serum lipid and lipoprotein levels and compositions appeared as early as 3 months after the alloxan injection and lasted for 12 months without any progress in the disturbance.

The expression of a membrane-associated lymphotoxin molecule (mLT) on lymphokine-activated killer (LAK) cells obtained from 18 patients with malignant tumors and its role in the tumor cell killing mechanisms were investigated. LAK cells from tumor-infiltrating lymphocytes (TIL-LAK cells) were mainly composed of CD3-positive cells, whereas LAK cells from peripheral blood lymphocytes (PBL-LAK cells) were mainly composed of CD16- and CD56-positive cells. However, mLT was found to be expressed on TIL-LAK cells as well as PBL-LAK cells. The degree of mLT expression correlated with the killing activity of LAK cells towards L929 cells (r = 0.806, P < 0.01, n = 15), but not with that towards Daudi or K562 cells. Although the degree of mLT expression correlated with the amount of secreted lymphotoxin (LT) in the supernatant of LAK cell culture, the secreted LT itself could not account for the tumor cell killing activity of LAK cells. Polyclonal rabbit anti-LT antibody partially inhibited the killing activities of LAK cells towards L929 cells and this inhibition was found in the combination of autologous tumor cells and PBL-LAK cells. These findings suggest the possibility that the mLT-related cytotoxicity is involved in the tumor cell killing mechanisms of TIL-LAK cells as well as PBL-LAK cells.

In an attempt to clarify the mechanisms of cytotoxicity mediated by CD4+ cytotoxic T lymphocytes (CTL), the expression of perforin and membrane-bound lymphotoxin (LT) (tumor necrosis factor-beta) in herpes simplex virus (HSV)-specific CD4+ human cytotoxic and noncytotoxic T- cell clones was examined. Three HSV-specific CD4+ human CTL clones that showed HLA-DR-restricted cytotoxicity and proliferative response were established. The cytotoxicity of these clones in 5-hour 51Cr release assays was found to be mediated by the directional target cell lysis and not by the release of cytotoxic soluble factors, ie, “innocent bystander” killing. Northern blot analysis showed that messenger RNAs for perforin and LT, which were both considered to be important mediators for cytotoxicity of CD8+ CTL and natural killer cells, were abundantly expressed in HSV-specific CD4+ CTL clones. Expression of perforin in the cytoplasm of CD4+ CTL clones was also detected by immunohistochemical staining using a monoclonal antibody against perforin. In addition, LT bound to the cell surface of CD4+ CTL clones was detected by flow cytometry. In contrast, little or no expression of perforin and LT was detected in three HSV-specific CD4+ noncytotoxic T- cell clones. Although the cytotoxicity mediated by lymphokine-activated killer cells was partly inhibited by addition of anti-LT antibody, it did not show any effect on the cytotoxicity of HSV-specific CD4+ CTL clones. In addition, it was found that cytotoxicity mediated by these CD4+ CTL clones was Ca2(+)-dependent. These data thus suggest that perforin and membrane-bound LT are both expressed in HSV-specific CD4+ CTL, although perforin might be the more important mediator in short- term culture.

The rat anti-mouse erythrocyte (MRBC) monoclonal antibody (mAb), R13, has been developed. The MRBC membrane protein recognized by R13 (R13-Ag) can be pirified by loading the butanol-extracted MRBC membrane solution on a R13-conjugated Cellulofine column in the presence of 0.1% CHAPS followed by elution with 1% CHAPS. The amino acid sequence of the affinity-purified R13-Ag corresponded to that predicted from the cDNA for the murine heat-stable antigen. It was revealed that the actual heat-stable antigen was composed of 27 amino acids.

Hyperphosphatasemia due to increased intestinal type serum alkaline phosphatase was noted in a 48-year-old male who had asymptomatic liver cirrhosis. The alkaline phosphatase activity in the serum was 828 U/l (our reference range in adults: 57–194 U/l), 94% of which was of the intestinal type as measured by an immunoprecipitation method. The intestinal component of alkaline phosphatase was separated into two major and some minor components using electrophoresis and isoelectrofocusing. One of the major components had similar mobility to that of a standard intestinal enzyme purified from adult intestine. The components were heat-labile and neuraminidase-resistant. Serial lectin affinity chromatography, however, indicated that sugar chain compositions of the alkaline phosphatase were different from those of the standard tissue intestinal enzyme. These results and further enzymological studies suggest that the patient's serum alkaline phophatase basically consisted of several intestine-like isoforms.

In order to clarify the relationship between composition and lipolytic responses to lipoprotein lipase (LPL), very low density lipoproteine (VLDL) from rats or humans were incubated with a commercially available LPL or with a partially purified LPL from postheparin human plasma and fatty acids released from VLDL were determined in vitro. VLDL from rats fed a diet containing 0.25 cholesterol for 6 months were rich in cholesterol and poor in triglycerides, and released less fatty acids from incubation with LPL than those from control rats. VLDL from normo- and hypertriglyceridemic human subjects were incubated with LPL. The fatty acid release poorly correlated with the apoprotein ratios of VLDL, apo C-III/C-II, B/E, and C/E with the exception of apo B/C, but it correlated well with the ratio of triglyceride/either one of the surface components including total apoproteins, free cholesterol and phospholipids in VLDL or the ratio of the triglyceride/total sum of the surface components. The correlation coefficients between fatty acid release and a ratio of triglyceride/total surface components were 0.774 (using the commercially available LPL) and 0.786 (using the partially purified human LPL). The fatty acid release increased after pretreatment of VLDL with phospholipase A2. The phospholipid content of VLDL was reduced without significant changes in other VLDL components. Thus, the responses of VLDL to LPL treatment may depend mainly upon the surface: core relationship of VLDL rather than its apoprotein composition except in rare clinical cases such as apo C-II deficiency.

Real-time virtual sonography (RVS) is a diagnostic imaging support system, which provides the same cross-sectional multiplanar reconstruction images as ultrasound images on the same monitor screen in real time. The purpose of this study was to evaluate radiofrequency ablation (RFA) assisted by RVS and CT for hepatocellular carcinoma (HCC) undetectable with conventional sonography. Subjects were 20 patients with 20 HCC nodules not detected by conventional sonography but detectable by CT or MRI. All patients had hepatitis C-induced liver cirrhosis; there were 13 males and 7 females aged 55-81 years (mean, 69.3 years). RFA was performed in the CT room, and the tumor was punctured with the assistance of RVS. CT was performed immediately after puncture, and ablation was performed after confirming that the needle had been inserted into the tumor precisely. The mean number of punctures and success rates of the first puncture were evaluated. Treatment effects were evaluated with dynamic CT every 3 months after RFA. RFA was technically feasible and local tumor control was achieved in all patients. The mean number of punctures was 1.1, and the success rate of the first puncture was 90.0%. This method enabled safe ablation without complications. The mean follow-up period was 13.5 month (range, 9-18 months). No local recurrence was observed at the follow-up points. In conclusion, RFA assisted by RVS and CT is a safe and efficacious method of treatment for HCC undetectable by conventional sonography.

The effect of ascorbic acid deficiency on serum and liver cholesterol, phospholipid and triglyceride levels, serum lipoprotein levels and serum lipoprotein cholesterol levels were examined in male rats with a hereditary defect in ascorbic acid synthesis (ODS rats). Male homozygotes (od/od) and male rats of their parent strain (+/+) were each divided into four treatment groups and were fed vitamin C-deficient or vitamin C-replete diets containing either 0 or 0.5% cholesterol. During the 3-wk feeding-period the ODS (od/od) rats fed the vitamin C-deficient diet gradually decreased food intake, resulting in a lower body weight than that of od/od rats given ascorbic acid. The serum cholesterol level was significantly higher in the vitamin C-deficient od/od rats fed the cholesterol diet, and it tended to be higher in those fed the control (0% cholesterol) diet, whereas the liver lipid levels remained unchanged relative to those in od/od rats fed the vitamin C-replete diet. The serum very low density lipoprotein and high density lipoprotein (HDL) cholesterol levels were lower in od/od rats fed the vitamin C-deficient diet without cholesterol, but intermediate density lipoprotein and low density lipoprotein cholesterol levels were markedly higher in the vitamin C-deficient od/od rats than in od/od rats given ascorbic acid, regardless of dietary cholesterol level. The ratio of HDL2 cholesterol to HDL3 cholesterol was also higher in the vitamin C-deficient od/od rats. The parent strain of the od/od rats (+/+) showed no change due to vitamin C deficiency. These results suggest that vitamin C deficiency delays low density lipoprotein metabolism and produces hypercholesterolemia in male od/od rats.

Cholesterol and bile acid leves were examined in young (8 weeks), middle-aged (12 months) and old (24 months) germ-free male rats, and young (8 weeks) and middle-aged (12 months) conventional male rats. The plasma cholesterol levels were higher in the aged rats, being more marked in the conventional rats. The liver cholesterol levels also increased with age and the increases were almost identical for both groups. No age-related changes were found in the biliary bile acid secretion, the pool size and distribution of bile acids in the bile, small intestine and large intestine, nor in the turnover frequency of bile acids, but the pool size in the young and middle-aged germ-free rats was much larger than that in the conventional rats. The turnover frequency was less in the germ-free rats. The bile acid synthesis presumed from the fecal bile acid excretion decreased in the aged germ-free rats but not in the conventional rats. A most remarkable age-related change was found in the bile acid composition; cholic acid increased and beta-muricholic acid derived from chenodeoxycholic acid in the rat decreased by aging, resulting in an increase of the CA/CDCA ratio (bile acids belonging to the cholic acid group/bile acids to the chenodeoxycholic acid group) in the bile, feces and pool. These results suggest that cholic acid synthesis increases while chenodeoxycholic acid synthesis is impaired by aging in rats.

Intense photostimulated luminescence (PSL) with a peak at 420 nm is observed in ultraviolet (UV) Iight-irradiated europium-doped potassium chloride (KCl: Eu) crystalline phosphors. The PSL characteristics of UV-irradiated KCl: Eu phosphor for optical memory application were studied. The excitation and emission mechanisms of the 420 nm PSL, which are consistent with the results obtained, are discussed.

Intense photostimulated luminescence (PSL) with a peak at 420 nm is observed in ultraviolet (UV)‐light‐irradiated europium‐doped potassium chloride (KCl:Eu) single crystals. The PSL characteristics of UV‐irradiated KCl:Eu for optical memory application are studied. The excitation and emission mechanisms of the 420 nm PSL, which are consistent with the results obtained, are discussed.

Serum cholesterol, triglyceride and phospholipid levels, liver cholesterol concentration, bile flow, biliary cholesterol, phospholipid and bile acid secretion rates, fecal sterol and bile acid levels and their bile acid compositions were examined in young-old parabiotic rats and compared with those in young and old control rats and young-young parabiotic rats. Bile acid composition was expressed in terms of the cholic acid group/chenodeoxycholic acid group (CA/CDCA) ratio. Body weight (BW) gain decreased after parabiosis especially in old rats, but the liver weight (g/100 g BW), diet-intake, feces dry weight, liver cholesterol concentration and fecal sterol level were almost the same in all the groups. The biliary bile acid secretion rate was higher and the fecal bile acid level was lower in old rats than those in young rats but both the levels became comparable with those in young rats after parabiosis of old rats with young rats. Young rats, however, showed no changes in these levels after parabiosis. The serum cholesterol level and the biliary and fecal CA/CDCA ratios in old rats were higher than those in young rats but decreased after parabiosis with young rats, although they were still higher than those in young rats. The serum cholesterol level in young rats increased after parabiosis with old rats, but not after parabiosis with young rats, and the fecal bile acid level and the CA/CDCA ratio were not changed in either case. It is concluded from these findings that the serum cholesterol level and the CA/CDCA ratio increased with age and that these increases were prevented after parabiosis with young rats, while young rats, although their serum cholesterol level was increased, showed no increase in the CA/CDCA ratio after parabiosis with old rats.

Pollen taxon in sediment samples can be identified by analyzing pollen morphology. Identification of related species based on pollen morphology is difficult and is limited primarily to genus or family. Because pollen grains of various ages are preserved at below 0 C in glaciers and thus are more likely to remain intact or to suffer little DNA fragmentation, genetic information from such pollen grains should enable identification of plant taxa below the genus level. However, no published studies have attempted detailed identification using DNA sequences obtained from pollen found in glaciers. As a preliminary step, this study attempted to analyze the DNA of Pinus pollen grains extracted from surface snow collected from the Belukha glacier in the Altai Mountains of Russia in the summer of 2003. A 150-bp rpoB fragment from the chloroplast genome in each Pinus pollen grain was amplified by polymerase chain reaction, and DNA products were sequenced to identify them at the section level. A total of 105 pollen grains were used for the test, and sequences were obtained from eight grains. From the sequences obtained, the pollen grains were identified as belonging to the section Quinquefoliae. Trees of the extant species Pinus sibirica in the section Quinquefoliae are currently found surrounding the glacier. The consistency of results for this section suggests that the pollen in the glacier originated from the same Pinus trees as those found in the immediate surroundings.

The effects of cholesterol feeding on serum and liver cholesterol levels, fecal and biliary bile acid levels, bile acid pool size and bile acid composition were examined in 2-, 12- and 24-month-old male germ-free rats. The major bile acids in these animals were cholic and β-muricholic acids. Cholesterol feeding increased synthesis of bile acids by 3- to 4-fold, especially that of chenodeoxycholic acid (mainly β-muricholic acid in the rat), decreasing the cholic acid/chenodeoxycholic acid (CA/CDCA) ratio in all rats regardless of age, even though the CA/CDCA ratio increased as a linear function of age in both diet groups. Cholesterol feeding increased the serum cholesterol level markedly in aged rats. This hypercholesterolemia may be produced by the increase in CA/CDCA ratio in aged rats.

Wistar male rats were treated for six days with broad spectrum beta-lactam antibiotics, latamoxef, and cefotaxime. On the seventh day, the number of fecal anaerobic microbes decreased, total fecal bile acids decreased, and bile acid pools increased. Secondary bile acids such as beta-hyocholic, hyodeoxycholic, lithocholic, and deoxycholic acids decreased in the feces while the primary bile acids, cholic, beta-muricholic, and chenodeoxycholic acids, became predominant. Coprostanol, a microbial metabolite of cholesterol, also disappeared from the feces during the treatment. The cecum enlarged to almost twice the size of that in control rats, whereas the liver weight was not significantly changed. After treatment was stopped, the number of fecal microbes returned to the initial counts within a week, but restoration of bile acid and cholesterol metabolism required at least three weeks.

Normal and alloxan diabetic rats were kept on a 0.25% cholesterol diet for 12 months and the changes in serum cholesterol levels, and fecal excretion of sterols and bile acids were examined to elucidate the influence of changes in bile acid metabolism on manifestations of hypercholesterolemia and development of atheromatous lesions. Diabetic rats fed the cholesterol diet showed increases in bile acid synthesis and in the cholic acid group/chenodeoxycholic acid group (CA/CDCA) ratio, and developed significant hypercholesterolemia and atheromatous lesions. In contrast, normal rats showed increased bile acids synthesis but a decreased CA/CDCA ratio after feeding with the cholesterol diet, and developed neither hypercholesterolemia nor atheromatous lesions. Fecal sterol excretion and the cholesterol/sitosterol ratio decreased in diabetic rats. Positive correlations were found between the cumulative serum cholesterol level and the atheromatous lesion area, and between the fecal CA/CDCA ratio and the serum cholesterol level, in the latter of which the correlation was higher in rats on the cholesterol diet than in those on the standard diet. These findings suggest that alteration of bile acid metabolism with increases in cholic acid synthesis and CA/CDCA ratio in diabetic rats enhances cholesterol absorption to produce significant hypercholesterolemia, which in turn leads to development of atheromatous lesions.

Bile acid synthesis from orally administered (14)C-cholesterol or from (14)C-mevalonic acid administered intravenously were examined in male and female 2-, 12- and 24-month-old rats. About 5-9% of the '4C-cholesterol administered was recovered as bile acids from the small intestine 3 days after administration. The value in the case of (14)C-mevalonic acid was about 1/10 that of (14)C-cholesterol. Major bile acids were cholic acid and beta-muricholic acid, and the formation ratio of cholic acid increased in aged rats, although no further difference was found between 12- and 24-month-old animals of either sex. Male rats formed more beta-muricholic acid than females at all ages examined. In addition, no significant difference was found in the bile acid composition between both precursors, (14)C-cholesterol and (14)C-mevalonic acid.

Intense optically stimulated luminescence (OSL) with a peak at about 420 nm is observed in ultraviolet(UV)-ray or X-ray irradiated europium-doped potassiumchloride (KC1:Eu) crystals. The OSL inytensity is increased with increasing UV—ray or X—ray irradiation dose. This suggests that KC1:Eu crystal is useful as a material for two-dimensional UV-ray or X-ray Imaging sensor utilizing OSL phenomenon. The results obtained are consistent with the proposed emission mechanisms of the L2O nm OSL peak, based on the recombination of electrons released from the F centers with Eu3' ions. The excitation mechanism for the OSL in UV-ray irradiated KC1:Eu crystals is also discussed. 1. INTRODUCTIONThe OSL phenomenon is based on the presence of electron and/or hole traps and luminescence centers in storage phosphor materials. During exposure toelectromagnetic waves such as ultraviolet(UV)-ray and X-ray, the traps are occupied with charged carriers such as electrons and holes that are created by UV—

The soluble form of guanylate cyclase, which is a heterodimer of two subunits with molecular weights of 82,000 and 70,000, was analyzed by radiation inactivation experiments to determine its functional size. Lyophilized crude extract from rat lung or the purified enzyme were irradiated with different doses from 60Co gamma-rays, and the residual activities were measured in the presence or absence of a potent activator, sodium nitroprusside. The target sizes for the basal activity and for the activity in the presence of sodium nitroprusside were calculated from the decay curve was 77 and 192 kDa, respectively, on the crude enzyme, or as 71 and 163 kDa, respectively, on the purified enzyme. The size for the activatable form of the enzyme was more than twice that of the basal activity and close to the size of the holoenzyme, implying that the enzyme activity must reside on one of the subunits and the activation by sodium nitroprusside requires interaction of both subunits.

Activation and inactivation of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase, which is a key enzyme of cholesterol synthesis, were examined in young and aged rats in vitro . The reductase activity in the liver microsomes of aged rats was always lower than that of young rats at various experimental conditions in vitro . But activation and inactivation of the reductase could operate on liver microsomes in aged rats as well as in young rats. Function of hepatic cytosol activator in aged rats seemed to be identical to that in young rats.

Four erythroleukemia cell lines were established in vitro from rats with primary leukemias induced by a set of intravenous injections of 7,8,12-trimethylbenz[a] anthracene (TMBA). These cell lines were grown in Dulbecco's medium with 20% inactivated fetal calf serum; they were characterized by specific consistent chromosome abnormalities, related to No. 2 chromosome, which were maintained in culture for more than 1 year. Addition of dexamethasone prevented the cell growth of 2 of the cell lines in this medium but failed to alter the growth rate of the other 2 cell lines. These findings are correlated with karyotype abnormalities involving chromosome No. 2.

Aged erythrocytes obtained by fractionation using gradient centrifugation with Dextran 40, showed lower glutathione reductase activity and riboflavin content than young cells. However, both young and old cells displayed almost the same increase in enzyme activity upon addition of flavin adenine dinucleotide to the test hemolysate in vitro. The lipid peroxide content in the cell membranes showed no consistent changes with aging.

Incorporations of squalene and mevalonate into cholesterol in hepatic microsomes and cytosol from growing or young rats were higher than those from adult rats. When the microsomes from growing or young rats were combined with the cytosol from adult rats instead of young cytosol, the incorporation rates of squalene and mevalonate into cholesterol were decreased. When adult microsomes were combined with young cytosol, cholesterol synthesis from mevalonate was increased but that from squalene was not. Cytosol fractions from cholesterol fed rats did not inhibit the cholesterol synthesis from squalene in the microsomes from non-treated rats, and the normal cytosol did not influence the cholesterol synthesis in the microsomes from cholesterol fed rats, suggesting the elevation of serum cholesterol in aged animals did not affect the activity of cytosol fraction. It is concluded that the function of hepatic sterol carrier protein is probably impaired by ageing process. The decrease of hepatic cholesterol synthesis in old microsomes in vitro is difficult to be recovered by the substitution of young sterol carrier protein for adult one. The decreased function of sterol carrier protein is not derived from hypercholesterolemia which is often observed in the aged.

Effects of sodium cholate, deoxycholate, chenodeoxycholate, and lithocholate on serum and liver cholesterol levels, bile flow, biliary cholesterol, phospholipids, and bile acids, and fecal sterols and bile acids were examined in Wistar strain male rats fed either an ordinary diet or a 2% cholesterol diet. Cholate and deoxycholate increased serum and liver cholesterol levels, serum pre beta-lipoprotein, bile flow, and biliary secretion of cholesterol, phospholipids, and bile acids, but chenodeoxycholate and lithocholate did not. The total amounts of sterols and of bile acids in the feces did not differ between the cholate and the chenodeoxycholate groups. All the bile acids except lithocholate decreased fecal coprostanol when the diet included cholesterol. Cholate and deoxycholate produced similar bile acid compositions in the bile and feces, as was the case between chenodeoxycholate and lithocholate, though chenodeoxycholate slightly increased the amount of muricholic acids, and lithocholate that of hyodeoxycholic acid, in the feces. The effects of cholate and deoxycholate are similar to each other but different from that of chenodeoxycholate or lithocholate in rats. Cholate causes marked accumulation of cholesterol in tissues, increased bile flow and biliary lipid secretion but chenodeoxycholate does not. Cholate is absorbed much more efficiently than chenodeoxycholate.

Stereological analysis at the electron microscope level was performed on zona fasciculata and zona reticularis of the adrenal cortex of male rats maintained on normal rat chow, clofibrate (hypolipidemic drug), cholesterol-rich diet or cholesterol-rich diet plus clofibrate. The purpose of this study is to know whether the alteration in the serum cholesterol level is accompanied by an alteration in the ultrastructure and steroidogenic activity of the adrenal cortex, and to compare these changes with those of Leydig cells reported previously. The corticosterone level in both serum and the adrenal tissue of cholesterol-fed rats remained unchanged. Treatment of the cholesterol-fed rats with clofibrate caused a significant rise in the serum corticosterone level but the concentration in the adrenal remained unchanged. Lipid droplets increased in number and volume significantly in both cortical zones in all experimental groups as compared with the control group. But neither cholesterol feeding nor clofibrate treatment of normal rats caused a change in the volume or surface area of smooth endoplasmic reticulum (ER). Treatment with clofibrate on the cholesterol-fed rats resulted in a decrease in the volume of smooth ER in both cortical zones. Mitochondria showed no change except for a decrease in volume and inner membrane surface area of zona reticularis of cholesterol-fed rats. Peroxisomes did not proliferate but rather decreased in clofibrate-treated rats. These changes in the adrenal cortex were different from those in Leydig cells. Possible reasons for these differences were discussed.

The normal levels of apolipoproteins in Japanese were investigated by the single radial immunodiffusion method. No differences were seen between the males and females in the apolipoprotein A-I, B, and C-III levels. However, the levels of apolipoproteins A-II and C-II were higher in the males, while that of apolipoprotein E was higher in the females. These differences were statistically significant. Except for apolipoprotein E, the apolipoprotein levels tended to increase with age more in the females than in the males. The apolipoprotein E level showed a statistically significant positive correlation not only with the total cholesterol level but also with the high density lipoprotein-cholesterol level.