Expression of luminescence in Photobacterium phosphoreum: Na+ regulation of in vivo luminescence appearance

In Photobacterium phosphoreum strain 496, growth and luminescence in a complex medium are optimal with 3% NaCl. However, in the same medium with 1% NaCl growth is similar, but the development of bioluminescence does not occur. In cells grown to mid or late-log phase in 1% NaCl, light emission can be triggered by the addition of NaCl, but the time required for its appearance is quite long, at least 30–45 min. The synthesis of m-RNA and protein are required for the development of luminescence, but the long time interval suggests that some intermediate steps are required. The time required is not less in conditioned 3% NaCl medium.     

A calcium-dependent 35-kilodalton substrate for epidermal growth factor receptor/kinase isolated from normal tissue

We have previously reported the isolation of a 35-kDa protein from A-431 cells that, in the presence of Ca2+, can serve as a substrate for the epidermal growth factor (EGF) receptor/tyrosine kinase (Fava, R.A., and Cohen, S. (1984) J. Biol. Chem. 259, 2636-2645). We now report the detection of an antigenically related 35-kDa protein in a number, but not all, of rat, pig, and human tissues. These antigenically related proteins also can serve as substrates for the EGF receptor/kinase in the presence of Ca2+. All of these proteins share the property of reversible, Ca2+-dependent binding to the particulate fraction (presumably membranes) of cell homogenates. We have isolated the 35-kDa substrate from porcine lung and have demonstrated that it is a Ca2+-binding protein. The amino-terminal sequence and the site of tyrosine phosphorylation therein have been determined. The positions of the acidic amino acid residues amino-terminal to the tyrosine phosphorylation site bear a distinct resemblance to the sequence in the homologous region of a number of other substrates for tyrosine kinases. Based on available data, the 35-kDa protein clearly differs from the protein I complex derived from intestinal mucosa and thought to be related to the proteins isolated herein (Gerke, V., and Weber, K. (1985) J. Biol. Chem. 260, 1688-1695). Finally, we report a striking sequence homology between the porcine 35-kDa described herein and human lipocortin, a phospholipase A2 inhibitor.     

Pepsin and autoimmune disease mouse

The effects of pepsin on autoimmune glomerulonephritis of MRL/1 mice were investigated. Intravenous administration of pepsin significantly improved survival rate and suppressed progressive increase in urinary excretion of protein and various histopathological changes in kidney. Increased serum levels of immune complex and anti-DNA antibody in MRL/1 mice were decreased by pepsin. Pepsin ameliorated abnormalities in lymphocyte subsets and lymphocyte functions. The fact that pepsin ameliorated abnormalities in immune function may contribute to the preventive effects of pepsin against pathogenesis and progress of immune complex nephritis.     

Structure of slow-reacting substance of anaphylaxis (SRS-A)

To elucidate the chemical structure of slow-reacting substance of anaphylaxis from rat (SRS-A^rat), SRS-A^rat were purified by the method of Orange with modification using DEAE-Sephadex A-25 chromatography. Ultraviolet absorption spectrum of purified SRS-A^rat indicated the presence of conjugated triene. Arylsulfatase B degradation products and HCl degradation products were subjected to analysis by a gas chromatography and mass spectrometry and a thin layer chromatography. Products obtained by arylsulfatase B catalysis contained 5,6-dihydroxy-7,9,11,14-eicosatetraenoic acid. HCl degradation products showed the presence of glycine, glutamic acid and cysteic acid. Furthermore, the analysis of anhydrous hydrazine degradation products of SRS-A^rat and of HCl hydrolyzed products of dinitrophenylated SRS-A^rat revealed the presence of glycine at C-terminal and glutamic acid at N-terminal. The study of the substrate specificity of arylsulfatase B against various materials including SRS-A^rat suggested the presence of sulfone in SRS-A^rat. The molecular ion peak of SRS-A^rat sodium salt was observed at m/e 680 in field desorption mass spectrum of SRS-A^rat.On the basis of these data, we identified the structure of SRS-A^rat as [γ-glutamyl-4(5-hydroxy-7,9,11,14-eicosatetraenoic acid-6-yl)-4,4-dioxocysteinyl] glycine.     

Contraction of seminal vesicle and prostaglandin E1

It was studied how PGE₁ would affect the responses of isolated human seminal vesicles to adrenalin. PGE₁ in the final concentration of 1.3 μg/ml suppressed the contraction of human seminal vesicle that would have occurred in reaction to adrenalin added one minute later. When the concentration of PGE₁ was increased to 6.7 μg/ml, the inhibitory action was further enhanced. The meanings of this phenomenon were discussed.     

Isolation and characterization of a plaque-forming lac-transducing phage phi80plac with special reference to its integration and excision

Summary From a double lysogen for 80dlac type II (Beckwith and Signer, 1966) and 80, we isolated a plaque-forming lac-transducing coliphage 80plac after selecting a strain with a suitable deletion in the 80 prophage. The lac region of the phage is i ⁺ o ⁺ z ⁺ y ⁺ a ^- and supposed to be located between genes 15 (N) and imm (CI). The phage showed feckless phenotype indicating deletion of genes of the red system. The phage is also deleted for int or att function, and integrates exclusively at the host lac region, largely dependent on the host rec system. Excision of the prophage upon UV-irradiation or by mating the male lysogen with a non-lysogenic female was efficient and largely dependent on the host rec system. But a considerable amount of rec-independent excision was observed at least in the case of zygotic induction, which was not likely to be caused by int-xis, red or ter system of the phage. 80plac/o ^e phage was also isolated by incorporation of o ^e1 mutation from strain 2000o ^e.     

Relationship between utilization of dual translational initiation signals and protein processing in Streptomyces

Summary Two sets of the Shine-Dalgarno sequence and the initiation codon (ATG) for translation of a gene encoding the protein SSI (Streptomyces subtilisin inhibitor) were studied in vivo by site-directed mutagenesis. The result shows that each ATG can function as an initiator of translation in either Streptomyces lividans 66 or Escherichia coli. The choice of initiation codon seems dependent on the host strain and is closely related to the processing mechanism of pre-SSI protein. The upstream ATG is presumed to be utilized preferentially giving two cleavage sites in pre-SSI in S. albogriseolus S-3253, the original SSI producer strain.Abbreviations SD Shine-Dalgarno - SSI Streptomyces subtilisin inhibitor     

The augmentation of tumor-specific immunity using haptenic muramyl dipeptide (MDP) derivatives

Summary Preinduction of potent haptenic muramyl dipeptide (MDP)-reactive helper T cell activity and subsequent immunization with MDP hapten-coupled syngeneic tumor cells resulted in enhanced induction of tumor-specific immunity through T-T cell collaboration between anti-MDP hapten helper T cells and tumor-specific effector T cells. The present study establishes two types of tumor-specific immunotherapy protocols utilizing helper T cells against MDP hapten cross-reactive with Bacillus Calmette Guérin (BCG). In the first model, naive normal C3H/He mice or mice in which MDP hapten-reactive helper T cells had been generated by BCG-sensitization were inoculated i.d. with syngeneic X5563 tumor cells. When both groups of mice were allowed to generate MDP hapten-modified tumor cells in the tumor mass in situ by intratumoral injection of MDP hapten, an appreciable number of growing tumors in the BCG-presensitized but not in the unsensitized group were observed to regress. In the second model, a growing X5563 tumor mass was removed by the surgical resection 9 days after the tumor implantation. Approximately 90% of C3H/He mice receiving such treatment died from tumor metastasis by about 30 days after the tumor resection. However, immunization of mice with MDP hapten-coupled X5563 tumor cells subsequent to the tumor resection resulted in an increased survival rate. Such protection from the tumor metastasis was appreciably stronger when compared to the protection obtained by immunization with MDP hapten-uncoupled tumor cells. The mice surviving in both models were also demonstrated to retain X5563 tumor-specific immunity. These results indicate that the presentation of MDP hapten-modified tumor cells to BCG-sensitized recipients results in potent tumor-specific immunity which contributes to the regression of the primary tumor or inhibition of metastatic tumor growth.This work was supported by a Grant-in-Aid for the Special Project Cancer Bioscience from the Ministry of Education, Science and Culture, Japan     

The growth analysis of water hyacinth,Eichhornia crasipes solms,in different water temperature conditions

A 40-day culture experiment of water hyacinth was made in 4 different water temperatures, 15, 20, 25 and 30°C, which were combined with 4 levels of concentration of culture solution, 1/3, 1, 3 and 9-fold of the standard solution containing 28 ppm of totalN and 7.7 ppm of totalP. The optimum condition for obtaining the maximum plant growth shifted from 30°C: 3-fold condition in the early stage to 20–25°C: 3-fold condition in the later stages of the experiment. The relation between the fresh weight biomass per 100-l tank,w, and the concentration of culture solution,f, was expressed successfully by a reciprocal equation,1/w=A F/f+A F ^′f/(1-f/C F)+B F, in whichA f,A f′, andB f are time dependent coefficients andC f is the upper limit of the concentration to permit plant growth which can change with time. The relation betweenw and water temperature,T, was expressed by another reciprocal equation,1/w=A T/e ^aT+A T^′e^bT+B T, in whicha andb are constants andAt At′ andB t are time dependent coefficients. The latter formulation shows that the temperature can be breated as an exponential factor, and it suggests the possibility of the growth coefficient of the logistic growth equation, ψ, being affected by temperature.     

Three distinct forms of atrial natriuretic factor receptors: kidney tubular epithelium cells and vascular smooth muscle cells contain different types of receptors

Three distinct ANF receptor subtypes have been identified and characterized from cultured canine kidney tubular (MDCK) cells and rat thoracic aortic smooth muscle (RTASM) cells. These three ANF receptor subtypes include; (1) a disulfide-linked 140 kDa protein found in RTASM cells which was reduced by sulfhydryl reagent dithiothreitol (DTT) to a 70 kDa band, (2) a disulfide-unlinked 120 kDa protein, specific to MDCK cells whose Mr was not reduced by DTT and (3) a 68-70 kDa protein prevalent in both RTASM and MDCK cells whose Mr was not reduced by DTT. The non-reducible 68-70 kDa and the reducible 140 kDa proteins showed strong affinities to the full-length ANF (99-126) and truncated ANF (103-123) peptides, however, non-reducible 120 kDa protein showed strong affinity only to the full length ANF (99-126) but negligible or very weak affinity to truncated ANF (103-123). These findings suggest that distinct ANF receptor subtypes are present in renal and vascular cells which might be linked to diverse physiological functions of ANF such as natriuresis and diuresis in kidney and vasorelaxation in vascular smooth muscle cells.