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We have studied water contained in the perfluorinated sulfonic acid (denoted as RSO3H) membrane by FT-IR spectroscopy. The IR spectra of the membrane, which absorbed water quickly even from the air, changed so drastically during dehydration that they showed clearly two isosbestic points in the 4000−1500 cm^-1 region. Analysis of the IR spectral changes made it clear that water molecules exist mostly as H3O⁺ in equilibrium with RSO3^- and RSO3H in the membrane. The IR change accompanying dehydration was found to result from the reaction as RSO3^- + H3O⁺ → RSO3H + H2O(evaporating out). The IR spectrum of H3O⁺ was clearly separated, which shows a broad OH stretching absorption with two unseparated peaks around 3370 and 3120 cm^-1 and an OH deformation absorption at about 1720 cm^-1. The spectrum is distinctively different from that of water separated, coexisting with H3O⁺, RSO3^-, and RSO3H in the membrane, which shows OH stretching and deformation absorptions at about 3480 and 1628 cm^-1, respectively.

Heparin-binding epidermal growth factor-like growth factor (HB-EGF) transduces mitogenic signals through the EGF receptor (EGFR). There are two forms of HB-EGF, the membrane-anchored form (pro-HB-EGF) and the soluble form (sHB-EGF). We studied the biological activity of pro-HB-EGF by using a model in which pro-HB-EGF-expressing effector cells was co-cultured with EGFR-expressing target cells. The DER cell, an EGFR-expressing derivative of the interleukin-3-dependent hematopoietic 32D cell line, grows well in the presence of EGF or sHB-EGF without IL-3. When DER cells were co-cultured on a monolayer of Vero-H cells overexpressing pro-HB-EGF, growth inhibition and subsequent apoptosis were induced in the DER cells even in the presence of excess amounts of EGF or sHB-EGF. Such growth inhibition of DER cells was abrogated when specific antagonists for pro-HB-EGF were added in the culture medium or when direct contact of DER cells with Vero-H cells was prevented, indicating that pro-HB-EGF is involved in this inhibitory effect. Pro-HB-EGF-induced apoptosis of DER cells was also observed even in the presence of IL-3. This rules out the possibility of simple competition between soluble EGFR ligands and pro-HB-EGF. Moreover, 32D cells expressing EGFR mutant composed of the extracellular and the transmembrane domain of EGFR and the cytoplasmic domain of erythropoietin receptor did not undergo apoptosis by co-culture with Vero-H cells, indicating that the inhibitory signal induced by pro-HB-EGF-expressing Vero-H cells is mediated to DER cells via EGFR and that the cytoplasmic domain of EGFR is essential for pro-HB-EGF-induced apoptosis. From these results, we concluded that pro-HB-EGF has unique biological activity through cell-cell contact that is distinct from the activity of sHB-EGF.

DRAP27, the monkey homolog of human CD9 antigen (DRAP27/CD9) and diphtheria toxin receptor (DTR) were expressed in mouse L cells. L cells transfected transiently with both DRAP27/CD9 and DTR cDNA bound approximately 10 times more diphtheria toxin (DT) than cells transfected with DTR alone. Stable L cell transfectants expressing both DTR and DRAP27/CD9 (LCH‐1 cells) had 15 times more cell surface DT‐binding sites and were 20 times more sensitive to DT than were stable L cell transfectants expressing DTR alone (LH‐1 cells). Increased DT‐binding and DT sensitivity were not due to increased DTR transcription or increased cell surface DTR protein. Co‐immunoprecipitation of DRAP27/CD9 with DTR and chemical cross‐linking suggest a tight association of these membrane‐bound proteins. In addition, the identity of DTR and a growth factor (HB‐EGF) was established. Immobilized DT specifically adsorbed HB‐EGF precursor solubilized from transfected L cells and [125I]DT bound to immobilized recombinant HB‐EGF. We conclude that DRAP27/CD9 associates tightly with DTR/HB‐EGF and up‐regulates the number of functional DTRs and DT sensitivity, and that HB‐EGF is identical to DTR.

A simple spectrophotometric method for the dectermination of D-glucosaminate with bacterial D-glaucosaminate dehydratase (EC 4.2.1.26) has been developed. The method is based on the enzymatic dehydration of D-gluco-saminatie and the spectrophotomeric determination of the product, 2-keto-3-deoxy-D-gluconate, as its semicarbazone. A linear relationship was established between the absorbance at 250 nm of semicarbasone and the amount of D-gluco-saminate (0.05 - 1.0 mol). The method was applicable to the determination of D-Glucosaminate in bacterial crude extracts.

DRAP27, the monkey homolog of human CD9 antigen (DRAP27/CD9) and diphtheria toxin receptor (DTR) were expressed in mouse L cells. L cells transfected transiently with both DRAP27/CD9 and DTR cDNA bound approximately 10 times more diphtheria toxin (DT) than cells transfected with DTR alone. Stable L cell transfectants expressing both DTR and DRAP27/CD9 (LCH‐1 cells) had 15 times more cell surface DT‐binding sites and were 20 times more sensitive to DT than were stable L cell transfectants expressing DTR alone (LH‐1 cells). Increased DT‐binding and DT sensitivity were not due to increased DTR transcription or increased cell surface DTR protein. Co‐immunoprecipitation of DRAP27/CD9 with DTR and chemical cross‐linking suggest a tight association of these membrane‐bound proteins. In addition, the identity of DTR and a growth factor (HB‐EGF) was established. Immobilized DT specifically adsorbed HB‐EGF precursor solubilized from transfected L cells and [125I]DT bound to immobilized recombinant HB‐EGF. We conclude that DRAP27/CD9 associates tightly with DTR/HB‐EGF and up‐regulates the number of functional DTRs and DT sensitivity, and that HB‐EGF is identical to DTR.

In vitro association of Xenopus nucleoplasmin and mammalian nonhistone chromosomal high mobility group 1 (HMG1) protein with nuclei isolated from rat liver was examined. Efficient association of nuclear proteins with isolated nuclei requires ATP, HCO3-, and Ca2+. Association occurred at 33 degrees C but not at 4 degrees C. ATP could be replaced by adenosine 5'-[alpha,beta-methylene]triphosphate (pp[CH2]pA), a nonhydrolyzable ATP analog. pp[CH2]pA associated with nuclei at 33 degrees C and nucleoplasmin and HMG1 rapidly associated with the pp[CH2]pA-bound nuclei at 4 degrees C. Competition studies showed that these associations at both 33 degrees C and 4 degrees C were specific. More than 80% of the bindings of nuclear proteins to the nuclear surface were blocked by wheat germ agglutinin.

Conventional solid-state ²⁷Al and ³¹P magic-angle spinning nuclear magnetic resonance (MAS NMR) and an advanced two-dimensional ²⁷Al multiple-quantum MAS NMR (2D ²⁷Al MQMAS NMR) have been applied to characterize MoO3−P2O5−Al2O3 hydrotreating catalysts. These catalysts were prepared by a sol−gel method with high Mo loadings (with Mo expected amounts of ~20 and ~30 wt %) and a wide range of P content (from 0 to 13 wt % of P). The chemical environments of Al and P strongly depend on the nature of the P precursor used in the sample preparation (H3PO4 or P2O5) and on the amount of P and Mo. From the ²⁷Al MAS NMR measurements on dried P-containing samples, formation of octahedral aluminum (Alocta) and of AlPO4 (Altetra−O−P surface species) in small amounts was observed. After calcination at 500 °C, the formation of tetrahedral aluminum (Altetra) and pentacoordinated aluminum (Alpenta) was also observed. The presence of molybdenum favored the interaction of phosphorus with the alumina framework, leading to significant amounts of Altetra−O−P. At higher P and Mo loadings, Al2(MoO4)3 was also detected in the calcined sample. ³¹P MAS NMR measurements revealed the formation of monomeric or polymeric P oxocompounds after drying and polymeric P oxocompounds or AlPO4 after calcination, respectively. The use of P2O5 as a P precursor gave rise to the formation of polymerized P oxocompounds rather than of AlPO4 after calcination in the absence of Mo. 2D ²⁷Al MQMAS NMR gave an improved resolution compared with conventional ²⁷Al MAS NMR. In dried samples, pure alumina and MoO3−alumina showed a single distorted Alocta site while P-containing catalysts showed additional octahedral and tetrahedral aluminum sites interacting with phosphorus (Alocta−O−P and Altetra−O−P, respectively). After calcination, mainly Alocta and Altetra sites with a small amount of Alpenta sites were observed in pure alumina. The presence of Mo introduced a large distortion in the alumina framework, since the amount of Alpenta significantly increased in the presence of Mo. Altetra−O−P sites were also detected in P-containing alumina catalysts, but the Alocta−O−P sites disappeared after calcination.

Temperature-programmed reduction (TPR) and temperature-programmed sulfiding (TPS) were used to characterize reduction and sulfiding properties of Fe-exchanged Y-zeolites and Fe-treated Y-zeolites, which were prepared by treating NH4Y-zeolite with an aqueous ferric nitrate solution (Fe-treatment). By considering their unique TPR and TPS patterns, it was confirmed that the Fe2+-species in the Fe-exchanged Y-zeolites are stabilized inside the sodalite cages and the hexagonal prisms. On the basis of the TPR and TPS characterizations, it was demonstrated that three types of the Fe-species are present in the Fe-treated Y-zeolites: ion-exchanged species, small Fe oxide clusters, and Fe oxide without interaction with the zeolite framework (including aggregated ferric oxide), the proportion of which is dependent on the extent of the Fe-treatment. Prolonged Fe-treatment weakens the interaction between the Fe-species and the framework oxygen atoms by hydrolysis, and leads to the aggregation of the Fe oxides and to the formation of bulk ferric oxide. The small Fe-oxide clusters, which are probably situated inside the supercages through a coordination with the framework oxygen atoms, are responsible for the high activity for toluene disproportionation in the presence of H2S.

Diphtheria toxin (DT) receptor associates with a 27-kD membrane protein (DRAP27) in monkey Vero cells. A cDNA encoding DRAP27 was isolated, and its nucleotide sequence was determined. The deduced amino acid sequence revealed that DRAP27 is the monkey homologue of human CD9 antigen. DRAP27 is recognized by CD9 antibodies. A human-mouse hybrid cell line (3279-10) possessing human chromosome 5, sensitive to DT, but not expressing CD9 antigen, was used for transfection experiments with DRAP27. When the cloned cDNA encoding DRAP27 was transiently expressed in 3279-10 cells, the total DT binding capacity was three to four times higher than that of untransfected controls. Transfectants stably expressing DRAP27 have an increased number of DT binding sites on the cell surface. Furthermore, the transfectants are 3-25 times more sensitive to DT than untransfected cells, and the sensitivity of these cells to DT is correlated with the number of DRAP27 molecules on the surface. However, when the cloned cDNA was introduced into mouse cell lines that do not express DT receptors, neither an increased DT binding nor enhancement of DT sensitivity was observed. Hence, we conclude that DRAP27 itself does not bind DT, but serves to increase DT binding and consequently enhances DT sensitivity of cells that have DT receptors. 12 proteins related to DRAP27/CD9 antigen were found through homology search analysis. These proteins appear to belong to a new family of transmembrane proteins.

Investigations were conducted with underyearling coho salmon Oncorhynchus kisutch reared in marine net-pens to determine the extent of genetic involvement in traits associated with smoltiffcation and saltwater adaptation and to assess genetic relationships among these traits. Data were obtained from two brood years (1977 and 1978) of experimental families formed by a nested mating design and consisted of length and weight measurements and morphological smolt assessments at 7 and 11 months postfertilization. Reasonably high heritability estimates (h2= 0.22–0.29) were obtained for all smoltification traits except survival (h2= 0.07). Correlation estimates among the variables suggested a strong genetic relationship between size and smoltification traits, with the exception of percent survival. The use of these estimates for genetic improvement was illustrated by two types of selection index. Substantial increases in smoltification and saltwater-adaptation traits appeared to be possible. Received April 25, 1983 Accepted March 20, 1984

Allele Frequency Distributions for Nine STR Loci in the Japanese Population

Heritability estimates and genetic and phenotypic correlations were calculated for weight and length of coho salmon during the freshwater period of growth. Estimates were based on measurements of progeny from 20 half-sib and 40 full-sib groups for two brood years. The heritability estimates for both brood years were high ranging from 0.61 to 0.22 and indicated that genetic improvement might be both rapid and substantial. Partitioning of the environmental variance component suggested that variation stemming from common environmental effects may be relatively unimportant even when full- and half-sib groups are reared together. Genetic and phenotypic correlations for weight and length of individual fish were high, inferring either pleiotropic gene action or close linkage of the associated genes. Values were consistent for the two brood years. Genetic correlations between other traits were significantly different from zero for one year class and nonsignifi cant, for the most part, in the other year class.