[Combination chemotherapy of MCNU and interferon alfa-2a (genetical recombination) for advanced multiple myeloma].
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Biomedical subjects
Publications and source records attributed to T Ono.
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The proliferating patterns of basal cell carcinoma were investigated by means of BrdU (bromodeoxyuridine), which is analogue of thymidine and by means of anti-BrdU monoclonal antibody. We classified 8 cases of basal cell carcinoma into three groups; 1) 21 nests of solid type basal cell carcinoma without retraction space 2) 6 nests of solid type basal cell carcinoma with retraction space 3) 5 nests of superficial type basal cell carcinoma without retraction space. The labeling index was obtained in each area of the outermost cells and inner cells in each nest of the three groups. The results indicated that when there was no retraction space, the labeling index of the outermost cells was high, and when there was some retraction space, the labeling index of the outermost cells was low. In other words, the proliferation of basal cell carcinoma cells occurs in the outermost cells of a nest. When there was retraction space, the role of the outermost cells of a nest diminished or disappeared.
Fatty acid-binding protein (FABP) was purified from rat gastric mucosa by successive Sephadex G-75 chromatography, DEAE-cellulose chromatography and HPLC on an RP-2 (Merck) reversed-phase column. The purified stomach FABP migrated as a single band corresponding to an apparent molecular mass of 15 kDa on SDS/PAGE. Stomach FABP appeared to be identical with rat heart FABP, as judged from its electrophoretic mobility, amino acid composition and tryptic peptide map. In addition, the amino acid sequences of two selected tryptic peptides coincided completely with the rat heart FABP sequence deduced from that of cDNA. Stomach FABP showed immunochemical identity with rat heart FABP when tested with an antiserum against rat heart FABP. Immunohistochemically, stomach FABP was specifically stained with anti-(rat heart FABP) serum in parietal cells of the gastric mucosa. The results suggested that the primary structure of stomach FABP is identical with that of rat heart FABP, and showed that stomach FABP is localized in parietal cells of the gastric mucosa.
A synthetic COOH-terminal oligopeptide of D1 protein deduced from the spinach psbA gene (Asn-325-Gly-353) was subjected to proteolytic digestion by purified processing enzyme of D1 protein [(1989) FEBS Lett. 246, 218-222] and the following two fragments were obtained as cleavage products: a COOH-terminal 9-amino-acid fragment (Ala-345-Gly-353) and an NH2-terminal 10-amino-acid fragment (Asn-325-Arg-334). It was concluded that: (i) the oligopeptide consisting of the COOH-terminal 29-amino-acid sequence deduced from the spinach psbA gene provides the recognition domain for the processing enzyme; (ii) the cleavage takes place at the predicted processing site of native D1 precursor protein (COOH side of Ala-344); and (iii) another cleavage takes place at an additional site (COOH side of Arg-334) for the synthetic substrate, but not for the native D1 precursor protein.
The complete primary structure of membrane-associated phospholipase A2 purified from a human splenic membrane fraction was determined by sequence analysis of the peptides generated by lysyl endopeptidase and Staphylococcus aureus V8 protease cleavage. The enzyme consists of 124 amino acid residues corresponding to a molecular weight of 13,904. The primary structure reveals the characteristics of Group II phospholipases A2 and a large ratio of basic amino acid residues to acidic ones, that ratio being 3.4 : 1.
Based on the partial amino acid sequences of membrane-associated phospholipase A2 (PLA2M), belonging to group II, purified from rat spleen, the cDNA encoding PLA2M was cloned by a new cloning strategy utilizing enzymatic cDNA amplification. At the N-terminus of the coded 146 residues, which were deduced from the cDNA sequence, the putative signal peptide was found despite the tight adherence of this enzyme to the membrane. The sequence of rat PLA2M exhibits 75% homology with that of human group II PLA2 in the protein-coding region. The result of RNA blot analysis showed that rat ileal mucosa contains the largest amount of the PLA2 transcript among the tissues examined.
Single unit activity in the lateral hypothalamus (LHA) of the rat was recorded while the animal learned to discriminate cue signals. Normally preferred potables (glucose, orange, or grape solution) or intracranial self-stimulation (ICSS) were used as rewards. Electric shock or tail pinch were used as aversive stimuli. The same behavior, licking, was the response required to either obtain the rewarding stimuli or avoid the aversive ones. For positive reinforcement a rat was rewarded with fluid or ICSS upon licking a spout presented in front of its mouth. In negative reinforcement experiments, an aversive stimulus, electric shock or tail pinch, was applied if the rat did not lick the spout. Solutions having smell only, taste only, or smell-plus-taste, were prepared from oranges or grape extract. Of 392 neurons analyzed, 256 responded differentially to rewarding and aversive stimuli, and 138 of these were tested with the 3 different solutions. Similar LHA neural responses occurred during actual drinking of the 3 kinds of solutions, as well as on recognition of the cue signal. Responses to smell only had shorter latency than responses to taste only. Neural activity in response to solutions that could be both smelled and tasted was the sum of activity in response to taste-only solutions plus that in response to smell-only solutions. Cue signal responses were rapidly acquired, usually within 2-5 trials, for both taste-only and smell-only solutions. The results indicate the integration of both taste and olfactory information by the same LHA neurons, and these neurons are involved in cue signal learning. Present results of LHA neuronal responses to taste and smell suggest that the intensity of gustation and olfaction may add together to enhance instinctive hedonic sensations. These neurons are involved in the formation of stimulus-reinforcement association in learning, and in elicitation of conditioned emotional responses.
Growth-stimulating activities of basic and acidic fibroblast growth factors (FGFs) toward human breast epithelial cells were examined and compared with the mitogenic activity of bovine pituitary extract (BPE) by the use of a serum-free medium which contained epidermal growth factor, insulin, transferrin, hydrocortisone, ethanolamine, phosphoethanolamine, prolactin and prostaglandin. Addition of 1 ng/ml of basic FGF (bFGF) to the serum-free medium significantly enhanced the growth potential of epithelial cells derived from human breast carcinoma, and the number of cells grown for 7 days with bFGF was more than 1 1/2 times higher than that in the serum-free medium containing BPE instead of prolactin and prostaglandin. Growth responsiveness toward bFGF of epithelial cells derived from histologically non-malignant breast tissues was lower than that of carcinoma-derived cells, and the growth-stimulating activity of bFGF was lower than that of BPE, which could significantly enhance the growth potential of the cells. Contrary to bFGF, acidic FGF at 1 ng/ml had no significant effect on the growth potential of breast epithelial cells which had grown out from either carcinoma or non-malignant tissues. The present results suggest that bFGF is a putative growth-stimulating factor for human breast epithelial cells, especially for carcinoma-derived cells, and can substitute at least in part for BPE in serum-free monolayer culture of the cells.
The amino acid sequence of rat heart fatty acid-binding protein was re-examined by analysing the tryptic and the chymotryptic peptides, since some discrepancies have been reported between the sequences determined by protein analyses and that deduced from the cDNA analyses. Our result completely agreed with the amino acid sequence predicted from the cDNA analyses, providing evidence for the actual existence of the molecular species predicted from the cDNA.
The properties of thromboxane A2 (TXA2) receptors were examined in 1321N1 human astrocytoma cells. 9,11-Epithio-11,12-methanothromboxane A2 (STA2), a stable analogue of TXA2, stimulated the accumulation of inositol phosphates (IPs) with an EC50 of about 50 nM. The STA2-induced accumulation of IPs was inhibited concentration dependently by ONO3708, a TXA2 receptor antagonist, with an inhibition constant (Ki) of about 10 nM. Inositol trisphosphate (IP3) was accumulated more rapidly than inositol bisphosphate (IP2) in response to STA2. HPLC analysis indicated that inositol 1,4,5-trisphosphate accumulated in the presence of STA2. STA2 alone had no effect on the accumulation of IPs in membrane preparations but it potentiated the accumulation induced by GTP gamma S. [3H]SQ29548, a TXA2 receptor antagonist, bound specifically to TXA2 receptors, expressing a single binding site with a dissociation constant (Kd) of 10.9 nM. The competition curve for STA2 inhibition of [3H]SQ29548 binding was shifted to the right and was steeper in the presence of GTP gamma S. Pertussis toxin (IAP) elicited ADP-ribosylation of 41KD protein but had no effect on the sensitivity to GTP of the STA2 inhibition of SQ29548 binding or of STA2-induced accumulation of IPs. It is concluded from these results that the stimulation of TXA2 receptors results in activation of phospholipase C via a GTP binding protein and that the protein is not a substrate for IAP.
A fatty acid-binding protein was purified from male rat kidney by the method of Fujii et al. [(1987) Arch. Biochem. Biophys. 254, 552-558]. Based on the close similarity of amino acid composition and the identity of the amino acid sequence in the amino-terminal region, the fatty acid-binding protein was identified as alpha 2U-globulin, which is the major male-specific protein in rat urine. The amino-terminal residue of the isolated kidney protein corresponds to residue 10 of alpha 2U-globulin, suggesting that the protein has been proteolytically processed in the kidney. This fatty acid-binding protein could not be immunologically detected in female rat kidney.
Calspermin is a heat-stable, acidic calmodulin-binding protein predominantly found in mammalian testis. The cDNA representing the rat form of this protein has been cloned from a rat testis lambda gt11 library. Sequence analysis of two overlapping clones revealed a 232-nucleotide 5'-nontranslated region, 510 nucleotides of open reading frame, a 148-nucleotide 3'-untranslated region, and a poly(A) tail. Authenticity of the clones was confirmed by comparison of a portion of the deduced amino acid sequence with the sequence of a tryptic peptide obtained from the rat testis protein. The lambda gt11 fusion protein was recognized by affinity purified antibodies to pig testis calspermin and bound 125I-calmodulin in a Ca2+-dependent manner. Calspermin cDNA encodes a 169-residue protein with a calculated Mr of 18,735. The putative calmodulin-binding domain is very close to the amino terminus of the protein. This region shows 46% identity with the calmodulin-binding region of rat brain Ca2+/calmodulin-dependent protein kinase II and 32% identity with the equivalent region of chicken smooth muscle myosin light chain kinase. The 5'-nontranslated region reveals significant homology with a portion of the catalytic region of the calmodulin-dependent protein kinase family. Calspermin contains a stretch of 17 contiguous glutamic acid residues in the central region of the molecule. Computer analysis predicts calspermin to be 81% alpha-helix and 14% random coil. Analysis of genomic DNA indicates calspermin to be the product of a unique gene. Northern blot analysis of rat testis RNA reveals a 1.1-kilobase mRNA. This RNA is restricted to testis among several rat tissues examined and could not be identified in total RNA isolated from testes of other mammals. Analysis of cells isolated from rat testis reveals calspermin mRNA to be predominantly expressed in postmeiotic cells indicating that it may be specific to haploid cells.
The content of mRNA for a pancreatic-type phospholipase A2 present in rat gastric mucosa was much greater than that in pancreas. In lung the mRNA for this pancreatic-type phospholipase A2 was also detected, but less than in pancreas. Nucleotide sequence analysis showed that these pancreatic-type phospholipase A2 cDNAs derived from rat gastric mucosa and lung were completely identical to that from rat pancreas (Ohara et al. (1986) J. Biochem. 99, 733-739). This demonstrates that the pancreatic-type phospholipase A2 present in gastric mucosa and lung does not originate from pancreas.
The rapid resolution of racemic N-4-nitrobenzoylamino acid isopropyl esters was accomplished without the loss of enantioselectivity by supercritical fluid chromatography (SFC) on novel chiral valine-diamide phases with carbon dioxide and a polar methanol modifier. In each stationary phase, a chiral moiety was anchored to the silica gel surface by a long decamethylene spacer. The enantioselectivity in SFC was comparable to that in liquid chromatography using 2-propanol-n-hexane. The time required for analysis was less than 5 min, and the range of enantiomer resolution (Rs) was 10.8-1.25. On using 2-propanol in place of methanol the separation was improved, but was accompanied by a decrease in column efficiency. The end-capping effect of the remaining surface silanols on enantiomer resolution is discussed.
Localization of hepatic fatty acid binding protein (h-FABP) in the small and large intestines of rats was studied by light and electron microscopic histochemistry using the peroxidase-antiperoxidase (PAP) method. In the small intestine of rats fed ad libitum, an intense FABP immunoreactivity was confined to the absorptive epithelial cells of the villi, but not of the crypts. The lowest margin of the immunoreactive cell sheet was closer to the crypts in the proximal than in the distal portions of the small intestine. In the large intestine, FABP immunoreactivity was present in the surface epithelial cells, with higher intensity in the proximal than in the distal portions of the intestine. After fasting rats for two days, many crypt cells exhibited intense immunoreactivity for h-FABP, resulting in an extension of the lowest margin of the immunoreactive cell sheet deep into the crypts. Such expansion of the immunoreactive cell population was reversed by refeeding the animals. With regard to the intracellular localization of immunoreactivity in the jejunum, the basolateral portion of the cytoplasm exhibited a more intense immunoreaction than the apical portion in the majority of immunoreactive cells lining the villi, whether the animals were fed or fasted. The immunoreactive products appeared in the cytoplasmic matrix without association with any subcellular structures.(ABSTRACT TRUNCATED AT 250 WORDS)
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Veillonellophage N2 adsorbed to polysaccharides (PSs) on Veillonella rodentium ATCC 17743 cell wall, and the bacteriophage receptor contained only glucosamine. D(+)-glucosamine hydrochloride (Sigma) also adsorbed the veillonellophage N2. These results therefore indicate that the receptor to the veillonellophage N2 is cell wall PSs. The PSs of the host cells as receptor have been characterized. Glucosamine accounted for approximately 100% of the weight of the PSs. The PSs which were partially resolved by Sephadex G-75 chromatography comprised approximately four glucosamine units. Their primary structure was determined by 400 MHz n.m.r. spectroscopy. One- and two-dimensional 1H-nmr experiments showed the PS to be a branched polymer. Glucosamine linkage was detected in one of the branches.
The effects of topically applied beta-cyclodextrin (beta-CyD) and heptakis(2,6-di-O-methyl)-beta-cyclodextrin (DM-beta-CyD) on photoallergic contact dermatitis due to chlorpromazine hydrochloride (CPZ) were investigated using the photomaximization technique in guinea pigs. From the gross and histological observations, the photosensitizing potential of CPZ with the simultaneous topical use of beta-CyDs was significantly lower than that of CPZ alone. The alleviating efficacy of DM-beta-CyD was greater than that of beta-CyD. beta-CyDs suppressed the penetration of CPZ into the skin through the formation of poorly skin-permeable complexes. In addition, beta-CyDs inhibited (a) the photoinduced free radicals derived from CPZ in the isolated dry skin and (b) the in vitro photochemical binding of CPZ to bovine serum albumin. These observations suggest that beta-CyDs suppress the photochemical reactions between CPZ and biological macromolecules present in the skin, resulting in the failure to form a photoantigen. The present results indicate that DM-beta-CyD is particularly effective in alleviating photoallergic contact dermatitis due to CPZ.