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The risk factors in pediatric influenza immediately before the COVID-19 era are not well understood. This study aims to evaluate the risk factors for hospitalization in pediatric influenza A and B for the recent seasons.Children with a fever of ≥38 °C and laboratory-confirmed influenza at 20 hospitals in outpatient settings in Japan in the 2013/14 to 2019/20 seasons were retrospectively reviewed. Possible risk factors, including gender, age, comorbidities, nursery school or kindergarten attendance, earlier diagnosis, no immunization, lower regional temperature, earlier season, and period of onset, were evaluated using binary logistic regression methods.A total of 13,040 (type A, 8861; B, 4179) children were evaluated. Significant risk factors (p 0.05) in multivariate analyses were young age, lower regional temperature, earlier season, respiratory illness (adjusted odds ratio [aOR]:2.76, 95% confidence interval [CI]:1.84-4.13), abnormal behavior and/or unusual speech (aOR:2.78, 95% CI:1.61-4.80), and seizures at onset (aOR:16.8, 95% CI:12.1-23.3) for influenza A; and young age, lower regional temperature, respiratory illness (aOR:1.99, 95% CI:1.00-3.95), history of febrile seizures (aOR:1.73, 95% CI:1.01-2.99), and seizures at onset (aOR:9.74, 95% CI:5.44-17.4) for influenza B.In addition to previously known factors, including young age, seizures, and respiratory illness, abnormal behavior and/or unusual speech and lower regional temperature are new factors. Negative immunization status was not a risk factor for hospitalization. A better understanding of risk factors may help improve the determination of indications for hospitalization during the future co-circulation of influenza and COVID-19.

SummaryAt the moment of union in fertilization, sperm and oocyte are transcriptionally silent. The ensuing onset of embryonic transcription (embryonic genome activation [EGA]) is critical for development, yet its timing and profile remain elusive in any vertebrate species. We here dissect transcription during EGA by high-resolution single-cell RNA sequencing of precisely synchronized mouse one-cell embryos. This reveals a program of embryonic gene expression (immediate EGA [iEGA]) initiating within 4 h of fertilization. Expression during iEGA produces canonically spliced transcripts, occurs substantially from the maternal genome, and is mostly downregulated at the two-cell stage. Transcribed genes predict regulation by transcription factors (TFs) associated with cancer, including c-Myc. Blocking c-Myc or other predicted regulatory TF activities disrupts iEGA and induces acute developmental arrest. These findings illuminate intracellular mechanisms that regulate the onset of mammalian development and hold promise for the study of cancer.

Various substrates have been used to stimulate habitat microbes in chloroethene-contaminated groundwater, however, the specific efficiency and minimum growth of microbes have rarely been studied. This study investigated the effects of seven substrates on trichloroethene (TCE) dechlorination by augmentation of groundwater with Dehalococcoides mccartyi NIT01 and its contribution to the microbial community. Three out of eight test groups completed dechlorination of 1 mM TCE-to-ethene in varying durations; groundwater supplemented with formate (FOR) required 78 days, whereas the microcosms with lactate (LAC) and citrate (CIT) required approximately twice as long (143 days). The calculated efficiency of how much produced H

This study presents the isolation of a novel strain of Dehalococcoides mccartyi, NIT01, which can completely dechlorinate up to 4.0 mM of trichloroethene to ethene via 1,2-cis-dichroroethene and vinyl chloride within 25 days. Strain NIT01 dechlorinated chloroethenes (CEs) at a temperature range of 25–32 °C and pH range of 6.5–7.8. The activity of the strain was inhibited by salt at more than 1.3% and inactivated by 1 h exposure to 2.0% air or 0.5 ppm hypochlorous acid. The genome of NIT01 was highly similar to that of the Dehalococcoides strains DCMB5, GT, 11a5, CBDB1, and CG5, and all included identical 16S rRNA genes. Moreover, NIT01 had 19 rdhA genes including NIT01-rdhA7 and rdhA13, which are almost identical to vcrA and pceA that encode known dehalogenases for tetrachloroethene and vinyl chloride, respectively. We also extracted RdhAs from the membrane fraction of NIT01 using 0.5% n-dodecyl-β- d -maltoside and separated them by anion exchange chromatography to identify those involved in CE dechlorination. LC/MS identification of the LDS-PAGE bands and RdhA activities in the fractions indicated cellular expression of six RdhAs. NIT01-RdhA7 (VcrA) and NIT01-RdhA15 were highly detected and NIT01-RdhA6 was the third-most detected. Among these three RdhAs, NIT01-RdhA15 and NIT01-RdhA6 had no biochemically identified relatives and were suggested to be novel functional dehalogenases for CEs. The expression of multiple dehalogenases may support bacterial tolerance to high concentrations of CEs.

Although the treatment of municipal wastewater using microbial fuel cells (MFCs) has been extensively studied, scaling the systems up for practical use remains challenging. In this study, a 226 L sewage treatment reactor was equipped with 27 MFC units, and its chemical oxygen demand (COD) removal and electricity production were evaluated. The MFC units were tubular air cores with a diameter of 5 cm and length of 100 cm, which were wrapped with a carbon-based cathode, anion exchange membrane (AEM), and nonwoven graphite fabric. The air-cathode–AEM MFC generated 0.12–0.30 A/m2, 0.072–0.51 W/m3, and 1.7–4.6 Wh/m3 in a chemostat reactor with a COD of 140–36 mg/L and hydraulic retention time (HRT) of 9–42 h throughout a year. The decrease in the COD was represented as the first-order rate constant of 0.038. The rate constant was comparable to that of other air-cathode MFCs with cation exchange membranes, indicating the necessity of a posttreatment to meet the discharge standard. It has been estimated that the MFC operation for 24 h before post-aeration can reduce the energy required to meet the discharge standard by 70%, suggesting the potential applicability of MFC in long HRT-treatments such as oxidation ditch. The resistances of the anode, cathode, and AEM were 0.60, 0.28, and 0.51 mΩ m2, respectively, and surface dirt rather than deterioration primarily increased the AEM resistance. A current exceeding 0.2 A/m2 significantly increases the anode potential, indicating that the current was limited by low COD. Increasing the anode-specific surface area can improve air–AEM MFCs used for practical applications.

The skin is responsible for a variety of physiological functions and is critical for wound healing and repair. Therefore, the regenerative capacity of the skin is important. However, stem cells responsible for maintaining the acral epithelium had not previously been identified. In this study, we identified the specific stem cells in the acral epithelium that participate in the long-term maintenance of sweat glands, ducts, and interadnexal epidermis and that facilitate the regeneration of these structures following injury. Lgr6-positive cells and Bmi1-positive cells were found to function as long-term multipotent stem cells that maintained the entire eccrine unit and the interadnexal epidermis. However, while Lgr6-positive cells were rapidly cycled and constantly supplied differentiated cells, Bmi1-positive cells were slow to cycle and occasionally entered the cell cycle under physiological conditions. Upon irradiation-induced injury, Bmi1-positive cells rapidly proliferated and regenerated injured epithelial tissue. Therefore, Bmi1-positive stem cells served as reservoir stem cells. Lgr5-positive cells were rapidly cycled and maintained only sweat glands; therefore, we concluded that these cells functioned as lineage-restricted progenitors. Taken together, our data demonstrated the identification of stem cells that maintained the entire acral epithelium and supported the different roles of three cellular classes.

Anaerobic mineralization of 2,4,6-tribromophenol (2,4,6-TBP) was achieved by a synthetic anaerobe community comprising a highly enriched culture of Dehalobacter sp. phylotype FTH1 acting as a reductive debrominator; Clostridium sp. strain Ma13 acting as a hydrogen supplier via glucose fermentation; and a novel 4-chlorophenol-degrading anaerobe, Desulfatiglans parachlorophenolica strain DS. 2,4,6-TBP was debrominated to phenol by the combined action of Ma13 and FTH1, then mineralized into CO2 by sequential introduction of DS, confirmed using [ring-(14)C(U)] phenol. The optimum concentrations of glucose, SO4(2-), and inoculum densities were 0.5 or 2.5mM, 1.0 or 2.5mM, and the densities equivalent to 10(4)copiesmL(-1) of the 16S rRNA genes, respectively. This resulted in the complete mineralization of 23μM 2,4,6-TBP within 35days (0.58μmolL(-1)d(-1)). Thus, using a synthetic microbial community of isolates or highly enriched cultures would be an efficient, optimizable, low-cost strategy for anaerobic bioremediation of halogenated aromatics.

Pentosidine (PEN), a well-defined advanced glycation end product (AGE), may be affected by psychological status, given the recent findings regarding AGE receptor functions. Because AGEs can be a factor in aging and in the development or worsening of many degenerative diseases, it is important to find a way to reduce the PEN levels in our body. This study aims to investigate novel factors associated with PEN levels.A cross-sectional study involving 106 female participants (aged 59-69) was conducted in a tea-producing district in Japan. The serum concentration of PEN was detected and evaluated in relation to the participants' psychological status, which was assessed using the Japanese version of the 28-item General Health Questionnaire (GHQ) and lifestyle factors. Factors that were significantly associated with PEN were analysed using multiple linear regression analyses. Significance was defined as p .05.The serum PEN concentrations were significantly and positively associated with the total GHQ scores and BMI after controlling for covariates (standardised beta coefficient (B) = 0.26, p .01; B = 0.27, p .01, respectively). In addition, the PEN levels were significantly lower in participants who consumed seven cups or more of green tea per day than those who consumed six or fewer cups per day (B = 0.19, p .05).Low GHQ scores (i.e. better psychological well-being) and green tea consumption may be helpful in decreasing AGEs.

Tetrabromobisphenol A (TBBPA) is the most widely used brominated flame retardant on the market. It has been detected in various environmental samples, and a growing body of evidence has demonstrated its toxic effects on living organisms. In this study, we report the enrichment and phylogenetic identification of bacteria that debrominate TBBPA to bisphenol A in the presence of humin. Incubation experiments indicated that humin was required for this debromination activity. Of the five compounds examined for inclusion in the TBBPA-debrominating culture, formate was the optimal electron donor. A 16S rRNA gene library showed that the culture was dominated by three known dehalogenator genera: Dehalobacter, Geobacter, and Sulfurospirillum. Further investigation indicated that Dehalobacter was responsible for the debromination of TBBPA. PCR-denaturing gradient gel electrophoresis analysis showed that Dehalobacter grew in the culture by utilizing TBBPA. Moreover, the copy number of the Dehalobacter 16S rRNA genes increased by about two orders of magnitude in the cultures without the addition of TBBPA, whereas it increased by approximately four orders of magnitude when TBBPA was present. The incubation experiments showed that Dehalobacter was reliant on humin for its debromination activity, indicating a new type of metabolism in Dehalobacter that is linked to humin respiration.

Biodegradable plastics (BPs) were evaluated for their applicability as sustainable and solid H(2) donors for microbial reductive dechlorination of 4,5,6,7-tetrachlorophthalide (fthalide). After a screening test of several BPs, the starch-based plastic (SP) that produced the highest levels of H(2) was selected for its use as the sole H(2) donor in this reaction. Fthalide dechlorination was successfully accomplished by combining an H(2)-producing SP culture and a KFL culture containing Dehalobacter species, supplemented with 0.13% and 0.5% SP, respectively. The efficiency of H(2) use in dechlorination was evaluated in a combined culture containing the KFL culture and strain Clostridium sp. Ma13, a new isolate that produces H(2) from SP. Results obtained with this culture indicated increased H(2)-fraction for fthalide dechlorination much more in this culture than in compared with a KFL culture supplemented with 20mM lactate, which are 0.75 H(2)·glucose(-1) and 0.015 H(2)·lactate(-1) in mol ratio, respectively.

Endogenous tocochromanols in extracted lipids from rice brans of the five cultivars were determined by high-performance liquid chromatography, and were investigated in relation to the fatty acid (FA) distribution of triacylglycerols (TAG) and phospholipids (PL). The dominant tocols were α-tocopherol and γ-tocotrienol, followed by α-tocotrienol and with much smaller amounts of γ-tocopherol and δ-tocotrienol. The lipids of these rice brans comprised mainly TAG (80.6-86.0wt.%), free FA (4.2-9.0wt.%), and phospholipids (5.5-6.7wt.%), whilst other components were also detected in minor proportions (0.2-2.1wt.%). The PL components included phosphatidyl choline (31.8-46.8wt.%), phosphatidyl ethanolamine (25.0-38.9wt.%) and phosphatidyl inositol (20.2-23.2wt.%). Comparison of these different cultivars showed, with a few exceptions, no significant differences (P>0.05) in FA distribution. FA distribution of TAG among the five cultivars was evident in the rice brans: unsaturated FA were predominantly concentrated at the sn-2 position and saturated FA primarily occupying the sn-1 or sn-3 position. These results suggest that the tocopherol content, lipid component, and FA distribution in rice brans are not dependent on the cultivation areas during the growing season.

A mass balance and kinetic investigation of anaerobic dechlorination of pentachlorophenol (PCP) was undertaken using an enriched microbial consortium in a laboratory scale continuous flow column, as a model microbial permeable reactive barrier. The chlorine balance showed that 50microM PCP was largely dechlorinated to phenol with the formation of a small quantity of 3-chlorophenol as an intermediate metabolite (hydraulic retention time 7.6days), and the chlorine removal efficiency reached 36microM d(-1). When the initial PCP concentration was increased to 100microM the chlorine removal efficiency increased 1.5 times. However, the dechlorination activity disappeared after 7.4 pore volumes (58days), demonstrating the susceptibility of the dechlorination culture to high concentrations of PCP. Lactate released hydrogen as an electron donor during PCP dechlorination, with acetate, propionate, CO(2) and CH(4) as byproducts. The carbon balance showed that some of the organic carbon source (PCP, lactate) in the influent was converted to gas and utilized for biomass growth in addition to organic metabolites. The kinetic study was conducted in a batch culture and yielded 1.99mg l(-1) biomass growth per unit of chlorine consumption (microM). The Monod equation was well fitted to the specific growth rate of 1.38d(-1) and a half saturation constant of 0.29microM. The organic chlorine removal rate in the batch culture was consistent with the results in the flow column, indicating the feasibility of and potential for in situ estimation and prediction through batch culture studies.

Fatty acid (FA) distributions and molecular species of triacylglycerols (TAG) isolated from total lipids extracted from adzuki beans ( Vigna angularis ) were analysed by a combination of AgNO 3 -TLC and GC, and were investigated in relation to the content of endogenous antioxidants determined by HPLC. δ-Tocopherol was present in the highest concentration (30.5 mg/kg beans), and γ-tocopherol in small amounts (12.8 mg/kg beans). The major lipid components were phospholipids (74.3%), TAG (13.5%), hydrocarbons (4.6%) and steryl esters (4.0%), whilst other components were also present in minor proportions (0.5–1.3%). Seventeen different molecular species were detected. The major TAG components were SMD (5.0%), S 2 T (19.2%), SD 2 (13.7%), SMT (9.3%), MD 2 (4.5%), SDT (7.0%), D 3 (8.8%) and ST 2 (15.8%) (where S, M, D, and T denote a saturated FA, a monoene, a diene, and triene, respectively). These results would be useful to both consumers and producers for manufacture of traditional adzuki foods in Japan.

Fatty acid distribution of triacylglycerols (TAG) and phospholipids (PL) obtained from the four cultivars of broad beans ( Vicia faba ) was investigated. Total lipids extracted from the beans were separated by thin-layer chromatography into eight fractions. The major lipid components were TAG (47.7–50.1%) and PL (47.5–50.5%), while hydrocarbons, steryl esters, free fatty acids, diacylglycerols (1,3-DAG and 1,2-DAG) and monoacylglycerols were present in minor proportions (1.8–2.4%). The major PL components were phosphatidylcholine (56.4–58.4%), phosphatidylethanolamine (20.3–21.7%) and phosphatidylinositol (16.6–18.6%). Phosphatidylinositol was unique in that it had the highest saturated fatty acid content among the three PL. No significant differences ( P > 0.05) in fatty acid distribution existed when the different cultivars were compared. The fatty acid distributions in the TAG were evident among the four cultivars: unsaturated fatty acids were predominantly concentrated in the sn −2 position, and saturated fatty acids primarily occupied the sn −1 or sn −3 position in the oils. These results could be useful to both consumers and producers for manufacture of traditional foods in Japan and elsewhere.

The anaerobic degradation of biphenyl was investigated in four uncontaminated Japanese paddy soils and one river sediment sample contaminated with benzene and chlorinated aliphatics. Two of the paddy soils and the sediment were capable of degrading biphenyl anaerobically without any additional medium or electron acceptors. The half-lives of biphenyl biodegradation in the three samples were 212 d in the Kuridashi soil, 327 d in the Kamajima soil, and 429 d in the river sediment. The Kuridashi soil metabolized 1+/-0.3% of [U-14C]-biphenyl into CO2 and 5+/-2% into water-soluble metabolites after 45 d of incubation. Submerged conditions, which result in lower nitrate and iron oxide contents, and neutral pH, appeared to be the common properties among the samples that influenced their degradation capacities. The addition of 10mM sulfate and 20mM Fe(III) as electron acceptors did not enhance the biphenyl degradation rate, whereas 10mM nitrate completely inhibited biphenyl degradation. The addition of different electron donors (lactate, acetate, or pyruvate) slightly slowed the degradation. Molybdate (an inhibitor of sulfate-reducing bacteria) had an inhibitory effect on biphenyl biodegradation, but bromoethanesulfonic acid (an inhibitor of methanogens) did not. Most biphenyl degradation was observed when only water was added, with no other electron acceptors or donors. These results suggest that sulfate-reducing bacteria and fermentative microbial populations play important roles in anaerobic biphenyl biodegradation in paddy soil.

Dechlorination of PCP has been observed previously under anaerobic condition in paddy soil. However, there is poor information about the dechlorination pathway of PCP and the microbial community associated with the PCP dechlorination in paddy soil. In this study, an anaerobic microbial community dechlorinating PCP was enriched by serial transfers from a paddy soil using a medium containing PCP, lactate and the steam-sterilized paddy soil. The enriched microbial community dechlorinated PCP completely to phenol under the anaerobic condition by a dechlorinating pathway as follows; PCP-->2,3,4,5-tetrachlorophenol-->3,4,5-trichlorophenol-->3,5-dichlorophenol-->3-chlorophenol-->phenol. Intermediate products such as 3-chlorophenol were not accumulated, which were immediately dechlorinated to phenol. The enriched microbial community was characterized physiologically by testing the effects of electron donors and electron acceptors on the dechlorinating activity. The dechlorinating activity was promoted with lactate, pyruvate, and hydrogen as electron donors but not with acetate. Electron acceptors, nitrate and sulphate, inhibited the dechlorinating activity competitively but not iron (III). The microbial group associated with the anaerobic dechlorination was characterized by the effect of specific inhibitors on the PCP dechlorination. Effects of specific metabolic inhibitors and antibiotics indicated the involvement of Gram-positive spore-forming bacteria with the PCP dechlorinating activity, which was represented as bacteria of phylum Firmicutes. The structure of the microbial community was characterized by fluorescence in situ hybridization, quinone profiling, and PCR-DGGE (denaturing gel gradient electrophoresis). The combined results indicated the predominance of Clostridium species of phylum Firmicutes in the microbial community. Desulfitobacterium spp. known as anaerobic Gram-positive spore-forming bacteria dechlorinating PCP were not detected by PCR using a specific primer set. These indicated a probable presence of novel anaerobic Gram-positive spore-forming bacteria dechlorinating PCP in the microbial community.

Sero-epidemiological studies are required to identify populations susceptible to measles. The hemagglutination inhibition (HI) test is no longer sensitive enough to confirm immunity to measles, and at present the particle agglutination (PA) test and enzyme-linked immunosorbent assay (EIA) are employed. The most reliable method is the neutralization test (NT), but it is time-consuming and requires experience. To simplify the NT, a recombinant measles AIK-C virus expressing green fluorescence protein (GFP-MVAIK) was constructed and used as a challenge virus. Plaques and cytopathic effects were visualized under ultraviolet light and detected easily, and measuring the intensity of the fluorescence enabled a reduction in the time-consuming steps. Neutralizing antibody titers of a complete inhibition neutralization test were equivalent to those of a 90% plaque reduction neutralization test. Comparison of four methods, HI, PA, EIA and the complete inhibition neutralization test, showed that only the results of EIA correlated well with those of the complete inhibition neutralization test, but sera with borderline levels by EIA were sometimes negative by the complete inhibition neutralization assay.

Terpenoids form a large and structurally diverse family of natural products and are ingredients of various herbal medicines. We have investigated possible interactions between herbal medicines and conventional medicines, and recently reported that monoterpenoids contained in Zanthoxyli Fructus can be potent inhibitors of P-glycoprotein (P-gp). In the present study, the influence of 70 kinds of terpenoids present in natural products on P-gp-mediated efflux transport was investigated. LLC-GA5-COL150 cells transfected with human MDR1 cDNA encoding P-gp were used to screen the terpenoids. Large increases in the intracellular accumulation of [(3)H]digoxin were observed in the presence of (R)-(+)-citronellal, (S)-(-)-beta-citronellol, alpha-terpinene, terpinolene, (-)-beta-pinene, abietic acid, ophiobolin A, cucurbitacin I, and glycyrrhetic acid. A study of the concentration-dependency revealed that the IC(50) of ophiobolin A, glycyrrhetic acid, (R)-(+)-citronellal, abietic acid, and cucurbitacin I was smaller than that of verapamil. The transcellular transport of [(3)H]digoxin across Caco-2 cell monolayers was then examined in the presence of (R)-(+)-citronellal, abietic acid, and glycyrrhetic acid. Significant increases in the apical-to-basolateral transport and decreases in the basolateral-to-apical transport and efflux ratio were demonstrated. These findings suggest that some natural products containing these terpenoids may inhibit P-gp-mediated transport and interact with P-gp substrates in the intestinal absorption process.

Complementary and alternative medicines can be applied concomitantly with conventional medicines; however, little drug information is available on these interactions. Previously, we reported on the inhibitory effects of an extract and monoterpenoids (e.g., (R)-(+)-citronellal) contained in citrus herbs on P-glycoprotein (P-gp) using P-gp-overexpressed LLC-PK1 cells. The objective of the present study was to investigate the effects of (R)-(+)-citronellal on P-gp-mediated transport in the intestinal absorption process in vitro and in vivo . Transcellular transport of [ 3 H]digoxin across Caco-2 cell monolayers was measured in the presence or absence of (R)-(+)-citronellal. (R)-(+)-citronellal reduced the basolateral-to-apical transport and efflux ratio for [ 3 H]digoxin significantly. Serum concentration-time profiles and pharmacokinetic parameters of digoxin after intravenous and oral administration were analyzed in rats pretreated with oral (R)-(+)-citronellal. The bioavailability of digoxin after oral administration decreased significantly to 75.8% of that after intravenous administration at the same dose. (R)-(+)-citronellal increased the bioavailability of oral digoxin to 99.9% but had no effects on total body clearance, volume of distribution, or elimination rate. These findings suggest that (R)-(+)-citronellal can increase the bioavailability of oral digoxin based on the blockade of P-gp-mediated efflux of digoxin from intestinal epithelia to the lumen in the absorption process. © 2006 Wiley-Liss, Inc. and the American Pharmacists Association.

Neuronal differentiation of embryonic neural progenitor cells is regulated by both intrinsic and extrinsic signals. Since dynamic changes in cell shape typify neuronal differentiation, cell adhesion molecules could be relevant to this process. Although it has been reported that fibronectin-integrin interactions are important for the proliferation of neural progenitor cells, little is known about the contribution of integrins to neuronal differentiation. In order to address this shortfall, we examined integrin expression on cortical progenitor cells by using immunohistochemistry and FACS analysis of cells in which GFP expression was driven by regulatory (promoter) regions of the nestin gene (nestin-GFP(+)). We here report that high levels of nestin promoter activity correlated with high expression levels of alpha(5)beta(1) integrin (alpha(5)beta(1)(high) cells). FACS analysis of nestin-GFP(+) cortical cells revealed an additional subpopulation with reduced expression of alpha(5)beta(1) integrin (alpha(5)beta(1)(low) cells). The size of the alpha(5)beta(1)(low) subpopulation increased during cortical development. To investigate the correlation between integrin and neuronal differentiation, nestin-GFP(+) cortical progenitor cells were sorted into alpha(5)beta(1)(high) or alpha(5)beta(1)(low) populations, and each potential to differentiate was analyzed. We show that the nestin-GFP(+) alpha(5)beta(1)(high) population corresponded to broadly multipotential neural progenitor cells, whereas nestin-GFP(+) alpha(5)beta(1)(low) cells appeared to be committed to a neuronal fate. These findings suggest that alpha(5)beta(1) expression on cortical progenitor cells is developmentally regulated and its downregulation is involved in the process of neuronal differentiation.

Hyaluronan (HA) associates with proteins and proteoglycans to form the extracellular HA-rich matrices that significantly affect cellular behaviors. So far, only the heavy chains of the plasma inter-α-trypsin inhibitor (ITI) family, designated as SHAPs (serum-derivedhyaluronan-associated proteins), have been shown to bind covalently to HA. The physiological significance of such a unique covalent complex has been unknown but is of great interest, because HA and the ITI family are abundant in tissues and in plasma, respectively, and the SHAP-HA complex is formed wherever HA meets plasma. We abolished the formation of the SHAP-HA complex in mice by targeting the gene of bikunin, the light chain of the ITI family members, which is essential for their biosynthesis. As a consequence, the cumulus oophorus, an investing structure unique to the oocyte of higher mammals, had a defect in forming the extracellular HA-rich matrix during expansion. The ovulated oocytes were completely devoid of matrix and were unfertilized, leading to severe female infertility. Intraperitoneal administration of ITI, accompanied by the formation of the SHAP-HA complex, fully rescued the defects. We conclude that the SHAP-HA complex is a major component of the HA-rich matrix of the cumulus oophorus and is essential for fertilizationin vivo.

The nuclear quadrupole resonance (NQR) spectra due to 35Cl, 81Br and 69Ga have been recorded for L2Ga2X6 [L  (CH3)4N and (C2Hs)4N; X  Cl and Br]. The NQR frequencies of the halogen in these compounds are considerably lower than those in Ga(III) compounds. The 69Ga NQR signals at 77 K are observed in the range 22–24 MHz. The electron distributions on the Ga and halogen atoms have been calculated according to the Townes-Dailey theory. The results show that the charge on each atom in the hexahalogenogallate(II) anions is independent of the cation but depends on the halogen.

Anomalous glycosylation shift values of the signals for C-1 (the anomeric carbon atom) and C-2 (the glycosyloxylated carbon atom) were sometimes observed for 2- O -β- d -glycopyranosyl (or -β- d -xylopyranosyl)-β- d -glucopyranosides, whereas no remarkable displacements of the other sugar-carbon signals were observed in these cases. This can be explained in terms of change of the orientation of the glycosyl linkages, owing to strong, steric interaction between the 2- O -glycosyl group and the 1- O -aglycon (or -sugar) group. Various 2- O -glycosylated α- and β- l -arabinopyranosides were synthesized. In the case of α- l -arabinopyranosides, the processes of 2- O -β- d -glucosylation, -β- d -xylosylation, and -α- l -arabinosylation resulted in unexpected, upfield shifts of the C-3, -4, and -5 signals, together with displacement of the C-1 and -2 resonances. Furthermore, significant alteration of the values of 3 J H1,H2 and 1 J C1,H1 was also observed for the 2- O -glycosylated α- l -arabinopyranoside moiety, indicating an increase in the contribution of the 1 C 4 conformation of the glycosylated α- l -arabinopyranoside in these cases. On the other hand, no remarkable variation in the signals of C-3, -4, and -5, or in the 3 J H1,H2 and 1 J C1,H1 values, was found for 2- O -α- l -rhamnosylation, except for 4-epihederagenin-3-yl 2- O -α- l -rhamnopyranosyl-α- l -arabinopyranoside. In the present study, such unusual 2- O -glycosylation shifts were not encountered for the 2- O -glycosylation of β- l -arabinopyranosides.