Molecular cloning and characterization of a novel salt-inducible gene encoding an acidic isoform of PR-5 protein in soybean (Glycine max [L.] Merr.).

We identified a novel salt-inducible soybean gene encoding an acidic-isoform of pathogenesis-related protein group 5 (PR-5 protein). The soybean PR-5-homologous gene, designated as Glycine max osmotin-like protein, acidic isoform (GmOLPa)), encodes a putative polypeptide having an N-terminal signal peptide. The mature GmOLPa protein without the signal peptide has a calculated molecular mass of 21.5 kDa and a pI value of 4.4, and was distinguishable from a known PR-5-homologous gene of soybean (namely P21 protein) through examination of the structural features. A comparison with two intracellular salt-inducible PR-5 proteins, tobacco osmotin and tomato NP24, revealed that GmOLPa did not have a C-terminal extension sequence functioning as a vacuole-targeting motif. The GmOLPa gene was transcribed constitutively in the soybean root and was induced almost exclusively in the root during 24 h of high-salt stress (300 mM NaCl). Interestingly, GmOLPa gene expression in the stem and leaf, not observed until 24 h, was markedly induced at 48 and 72 h after commencement of the high-salt stress. Abscisic acid (ABA) and dehydration also induced expression of the GmOLPa gene in the root; additionally, dehydration slightly induced expression in the stem and leaf. In fact, the 5'-upstream sequence of the GmOLPa gene contained several putative cis-elements known to be involved in responsiveness to ABA and dehydration, e.g. ABA-responsive element (ABRE), MYB/MYC, and low temperature-responsive element (LTRE). These results suggested that GmOLPa may function as a protective PR-5 protein in the extracellular space of the soybean root in response to high-salt stress and dehydration.

A preliminary description of the crystal structure of gamma-glutamyltranspeptidase from E. coli K-12.

The structure of GGT [EC 2.3.2.2] from E. coli K-12 was studied at 3 A resolution by X-ray crystallography. Initial protein phases were calculated using two kinds of Pb2+ derivatives. The phases were refined by non-crystallographic 2-fold symmetry electron density averaging combined with solvent flattening and histogram matching. The GGT molecule has overall dimensions of 60 x 50 x 40 A. There are two antiparallel beta-pleated sheets consisting of 6 and 7 beta-strands. The two beta-sheets form a wall-like structure. Twelve short alpha-helices were detected, of which the maximum length appears to be four helix turns.

Subunit association of gamma-glutamyltranspeptidase of Escherichia coli K-12.

gamma-Glutamyltranspeptidase [EC 2.3.2.2] of Escherichia coli K-12 consists of one large subunit and one small subunit, which can be separated from each other by high-performance liquid chromatography. Using ion spray mass spectrometry, the masses of the large and the small subunit were determined to be 39,207 and 20,015, respectively. The large subunit exhibited no gamma-glutamyltranspeptidase activity and the small subunit had little enzymatic activity, but a mixture of the two subunits showed partial recovery of the enzymatic activity. The results of native-polyacrylamide gel electrophoresis suggested that they could partially recombine, and that the recombined dimer exhibited enzymatic activity. The gene of gamma-glutamyltranspeptidase encoded a signal peptide, and the large and small subunits in a single open reading frame in that order. Two kinds of plasmid were constructed encoding the signal peptide and either the large or the small subunit. A gamma-glutamyltranspeptidase-less mutant of E. coli K-12 was transformed with each plasmid or with both of them. The strain harboring the plasmid encoding each subunit produced a small amount of the corresponding subunit protein in the periplasmic space but exhibited no enzymatic activity. The strain transformed with both plasmids together exhibited the enzymatic activity, but its specific activity was approximately 3% of that of a strain harboring a plasmid encoding the intact structural gene. These results indicate that a portion of the separated large and small subunits can be reconstituted in vitro and exhibit the enzymatic activity, and that the expressed large and small subunits independently are able to associate in vivo and be folded into an active structure, though the specific activity of the associated subunits was much lower than that of native enzyme. This suggests that the synthesis of gamma-glutamyltranspeptidase in a single precursor polypeptide and subsequent processing are more effective to construct the intact structure of gamma-glutamyltranspeptidase than the association of the separated large and small subunits.

A high dose chemoradiotherapy and peripheral blood stem cell support combined with the CD34(+)-selection method in cyclin D1(+)-mantle cell lymphoma.

Mantle cell lymphoma (MCL) is currently regarded as one of the most incurable lymphomas, although reliable prognostic indicators are not yet to be defined. In a previous report, it was indicated that most of the patients with immunohistochemically cyclin D1(+)-MCL pursued the lethal clinical course within 7 years, not having achieved complete remission (CR). Recently, a high dose chemoradiotherapy was carried out, this was supported by peripheral blood stem cell transplantation (PBSCT) using the CD34(+)-selection method in a 48-year-old female patient with cyclin D1(+)-MCL. The tumor cells were detected in her peripheral blood despite four courses of combination chemotherapy using CHOP regimen. Soon after the pre-conditioning of total body irradiation (TBI) and high dose melphalan, she received the PBSCT of 1.8 x 10(6)/kg CD34+ cells and showed rapid hematological recovery without life-threatening complications. The patient achieved CR and was alive, without disease, 730 days after PBSCT. Thus, CD34+ selected PBSCT appears to provide further insight into the effective treatment and possible cure of this aggressive disease, i.e. cyclin D1(+)-MCL.

Colliding malignant lymphoma and adenocarcinoma of the stomach: case report.

We report a case of malignant lymphoma presenting as the superior vena cava syndrome. This case is very interesting since both adenocarcinoma mucocellulare scirrhosum, Borrman type IV, and lymphoma were present in the stomach, forming colliding neoplasms. Rather than systemically disseminated, the lymphoma cells were localized in the stomach, the intestine, a few perigastric lymph nodes and in a mediastinal mass, which may have constituted a neoplastic thymus invaded by lymphoma cells from the stomach. The unique occurrence of the histological combination as gastric colliding neoplasms is discussed.

Quantitative evaluation of selenium contained in tea by high performance liquid chromatography.

For determination of selenium (Se) in biological materials, an improved method based on high performance liquid chromatographic determination of the fluorophore formed by reaction of selenite with 2,3-diaminonapththalene was developed. The concentration detection limits were 0.5 ng/g in dried materials and 0.03 ng/mL in fluid materials. In quadruplicate assays of 11 biological reference materials using the proposed method, measured Se concentrations were not significantly different from their certified values. Thus, the proposed method is reliable and suitable for the determination of trace levels of Se in foods. Using the proposed method, Se concentrations in various kinds of tea were determined to assess the contribution of tea to daily Se intake in the Japanese population. Se concentration in the leaves of general black, green and oolong tea obtained in local retail stores was 33 +/- 19 ng/g (n=440). The leaves of a particular Chinese green tea sold under the name "high Se tea" were found to contain 455 +/- 184 ng/g (n= 14) of Se. While the percentage of Se extractable by infusion was less than 5% for the general teas, that in the high Se tea was more than 20%. These results indicated that intake of tea does not contribute to daily Se intake in the Japanese population. However, since infusions from high Se tea contained over 5 ng/mL of Se, consumption of over 1 L/d of tea derived from such high Se teas may increase the daily Se intake by close to 10%.

Comparison of the inhibitory effect of sulfamonomethoxine and other sulfonamides on capsule formation of Bordetella bronchiseptica.

The inhibitory effect of sulfamonomethoxine and other sulfonamides on the capsule formation of sulfonamide-resistant Bordetella bronchiseptica was investigated. All the sulfonamides having MeO(-OCH3) groups inhibited the capsule formation of B. bronchiseptica. Strong inhibition was obtained with sulfamonomethoxine. Inhibition was not seen with sulfonamides having no MeO groups.

Clinical investigation of vestibular damage by antituberculous drugs.

Vestibular function testing was done regularly on the cases given streptomycin, kanamycin, or enviomycin and a method to detect the cases of vestibular dysfunction at an early stage was discussed, as well as the time these drugs should be discontinued. Subjects were 85 cases of tuberculosis treated with streptomycin, kanamycin, or enviomycin who were admitted to our hospital from December 1984 to May 1986. The method of equilibrium examination performed at regular intervals is as follows: standing test (Romberg test), stepping test, and Meyer zum Gottesberge's head-shaking test were done once a week for a month after starting antituberculous injections and they were re-examined once every 2 weeks for at least 3 months after beginning the injections. After the 3 months these tests were done once a month. Eight cases of vestibular damage due to streptomycin or enviomycin could be easily detected at an early stage by performing Meyer zum Gottesberge's head-shaking test, together with the standing test and the stepping test. Vestibular dysfunction is apt to occur after about 1 month or within a month from the start of daily injections especially with streptomycin. Therefore, the method of equilibrium examination, we suggest, is that the Meyer zum Gottesberge's head-shaking test, the standing test (Romberg test), and the stepping test should be performed once a week during the first month after the start of this drug. When the result of the Meyer zum Gottesberge's head-shaking test is less than 50% and swaying and/or rotation occur in the stepping test, the drugs being given should be discontinued.

[Serum adenosine deaminase (ADA) activity in patients with active pulmonary tuberculosis].

To clarify the immunological aspect of tuberculosis, we investigated serum adenosine deaminase (ADA) activity, T and B cell percentile in total peripheral mononuclear cells, peripheral T cell subpopulation and their relationship with other inflammatory parameters in 20 patients with active pulmonary tuberculosis. Serum ADA activity showed abnormal high level in all patients in nontreated phase with significant regression after three months treatment by anti-tuberculous drugs (P less than 0.05). In addition, significant positive correlation was observed between serum ADA activity and erythrocyte sedimentation rate at 1 hour (r = 0.56). Concerning about peripheral T cell subpopulation studied by two colour flow cytometry with FITC-conjugated anti-CD4 and phycoerythrin-conjugated anti-HLA-DR antibody, there was positive correlation between CD4(-) HLA-DR(+) T cell and serum ADA activity (r = 0.59) without any abnormal frequency of each T cell subpopulation. Additionally, after the treatment of pulmonary tuberculosis, significant increase of T cell percentile in total peripheral mononuclear cells were observed (P less than 0.005). In conclusion, these results may suggest serum ADA activity could be a parameter of activity of pulmonary tuberculosis and reflect the function of activated suppressor/cytotoxic T cell.

[Primary pulmonary amyloidosis with multiple nodules].

A 42-year-old man complained of a productive cough and exertional dyspnea. Roentgenogram and computed tomography of the chest showed several nodular shadows, and an open lung biopsy was done. Congo-red staining and electron microscopy showed the pulmonary nodules to be amyloid with infiltration by plasma cells and foreign body giant cells. The amyloid was identified as AL-lambda and kappa type by immunohistochemical examination, and plasma cells and foreign body giant cells contained light chains. In situ hybridization studies showed intens signals of lambda mRNA and kappa mRNA in the plasma cells, but no signal in the foreign body giant cells. The foreign body giant cells appeared to have phogocytosed the light chains. Electron microscopic examination showed an "amyloid star" in the foreign body giant cells. Morphologically, this observation showed that the amyloid was produced by the foreign body giant cells.

Sjögren's syndrome with bronchial gland involvement and multiple bullae.

A 52-year-old woman with Sjögren's syndrome presented with a dense lymphocytic infiltrate around the bronchial gland disclosed by a transbronchial lung biopsy specimen. Two and a half years after the initial evaluation, her pulmonary status had deteriorated and multiple bullae had developed at both lung bases.

Occurrence of coagulase serotype among Staphylococcus aureus strains isolated from healthy individuals--special reference to correlation with size of protein-A gene--.

One-hundred-and-nineteen strains of Staphylococcus aureus isolated from healthy individuals for 3 years between 1991 and 1993 were subjected to an investigation on the producibility of proteins including protein A, coagulase, enterotoxins and toxic-shock syndrome toxin-1. Especially, protein A was the center of our interest. Among these strains, 69, 43, 3 and 1 strains were found to have the protein-A gene containing 5, 4, 3 and 2 IgG-binding domains, respectively. On the other hand, only one strain was devoid of the protein-A gene. There were some differences in the profile of the coagulase serotype between the group with 4 IgG-binding domains and that with 5 IgG-binding domains. Differences in the profile of toxin production were also observed between the two groups.