The effect of heptakis (2,6-O-dimethyl) beta-cyclodextrin (Me beta CD) on the production of pertussis toxin was evaluated. The addition of Me beta CD to the medium stimulated cell growth and pertussis toxin production. Me beta CD enhanced pertussis toxin production 100 times more in synthetic media, such as Stainer-Scholte medium (D. W. Stainer and M. J. Scholte, J. Gen. Microbiol. 63:211-220), than in Me beta CD-free medium in 2-day shake cultures. Maximum production of pertussis toxin was estimated as 50 mg of protein per liter of culture broth both by in vitro and in vivo assays. Purified toxin was demonstrated to be biochemically and biologically identical to the toxin produced in Me beta CD-free static cultures.

Bacterial Toxins↗

PubMed1983

Heptakis(2,6-O-dimethyl)beta-cyclodextrin: a novel growth stimulant for Bordetella pertussis phase I.

The effect of cyclodextrins on the growth of Bordetella pertussis Tohama phase I in synthetic medium was evaluated. The addition of cyclodextrins, especially heptakis(2,6-O-dimethyl)beta-cyclodextrin (Me beta CD), to a complete synthetic medium such as Stainer-Scholte medium gave the same number of individual colonies and growth rates as those on Bordet-Gengou medium. Furthermore, with the addition of Me beta CD, growth inhibition by fatty acids such as oleic or palmitic acid was overcome and normal cell growth was observed. This modified Stainer-Scholte medium, designated as cyclodextrin solid medium (CSM), supported excellent growth of 20 lyophilized clinical isolates. Serotypes of the organisms after 10 passages on this CSM plate were not changed. These results suggest that Me beta CD is a significant growth stimulant and CSM is one of the most suitable synthetic media for culture of B. pertussis phase I.

Antigens, Bacterial↗

PubMed1983

Riboflavin metabolism in the single lens of rat.

To study the mode of riboflavin metabolism in the lens in vivo, the uptake of 14C-labeled riboflavin and synthesis of ester forms from 14C-riboflavin were assayed in the single whole lens of the rats. 14C-riboflavin uptake increased in proportion to the substrate concentration in the incubation medium, reaching a plateau within 60 min. At 4 degrees C the uptake was lowest, at 30, 35 and 40 degrees C it was similar. Neither uncoupler (2,4-dinitrophenol and monoiodoacetate) nor ouabain inhibited the uptake. Within 15 min, approximately 35% of transported 14C-riboflavin was converted to ester forms of riboflavin; this ratio remained constant until 120 min of incubation. Characteristically, 2-3% of 14C-riboflavin was bound to lens protein. 2,4-Dinitrophenol or monoiodoacetate inhibited the synthesis of ester forms of riboflavin, but ouabain did not. No synthesis of ester forms of riboflavin was observed in the lens capsule with epithelium. These results suggest that two enzymes, flavokinase and flavin adenine dinucleotide (FAD) pyrophosphorylase which convert riboflavin to flavin mononucleotide and FAD, respectively, exist in the lens of the rat. These enzymes may participate in riboflavin uptake and riboflavin may pass through the lens capsule by simple diffusion without undergoing a conformational change and then may be metabolized to ester forms of riboflavin with subsequent partial binding to lens protein.

Animals↗

PubMed1983

Coronary angiographic findings in various types of unstable angina--study on the pathophysiology of unstable angina.

Coronary angiographic findings were studied in 129 patients with various types of unstable angina in order to clarify the pathophysiology of unstable angina. The subjects were divided into 3 types: effort angina (E), rest angina (R), and effort and rest angina (E+R), and each of these 3 types was subdivided into group I (new onset), II (recurrent) and III (changing pattern). 1) R had less severe coronary lesions than E or E+R. 2) Severity and distribution of coronary atherosclerotic lesions in unstable angina were similar to those in stable angina. 3) Incidence of coronary spasm is higher in unstable R and E+R than in stable R and E+R, respectively. 4) Unstable R and E+R with frequent attacks were associated with a higher frequency of coronary spasm and severe proximal coronary stenosis than those without frequent attacks, respectively. 5) Among unstable E+R-III (changing pattern), the patients who developed E+R from E showed significantly higher incidence of multiple vessel disease than those who developed E+R from R and significantly lower incidence of spontaneous spasm than those with E+R, who remained with the same pattern but in whom the frequency and/or the intensity of the attack increased, without any significant difference in the severity of coronary stenosis from other 2 subgroups. It is concluded that coronary spasm as well as severe coronary atherosclerotic lesions may be responsible for the unstable state of angina. Especially in R and E+R, coronary spasm is the most important factor responsible for the unstabilization of angina.

Adult↗

PubMed1983

Characterization of testosterone binding protein of rat liver cytosol.

Testosterone binding protein from rat liver cytosol, which had been incubated with [3H]testosterone followed by treatment with dextran-coated charcoal, was analyzed by DEAE-cellulose and phosphocellulose chromatography. On DEAE-cellulose chromatography, two distinct peaks of radioactivity were eluted at 0.07 M and 0.19 M KCl, both sedimented in 4 S regions. Phosphocellulose chromatography resulted in a broad peak at 0.08 M KCl, with a shoulder at 0.04 M KCl, both sedimented at 4 S. These findings indicated that testosterone binding protein consists of two types of components each with 4 S.

Androgen-Binding Protein↗

PubMed1983

Functional heterogeneity of the Lyb-5- B cell subpopulation: mutant xid B cells and normal Lyb-5- B cells differ in their responsiveness to phenol-extracted lipopolysaccharide.

In the present study, responses stimulated by phenol-extracted lipopolysaccharide (LPS(phenol)) and butanol-extracted LPS (LPS(butanol)) were used to assess the possibility that xid B cells might not be identical to the Lyb-5- B cells present in normal mice. It was found that xid B cells responded well only to LPS(butanol) whereas normal B cells responded well to both LPS(butanol) and LPS(phenol). Thus, LPS(butanol) appeared to be a TI-1 antigen and LPS(phenol) appeared to be a TI-2 antigen. In contrast to classical TI-2 responses, however, responses stimulated by LPS(phenol) did not exhibit a stringent requirement for accessory cells. Furthermore, if LPS(phenol) were a classical TI-2 antigen, it should only activate Lyb-5+ B cells. To determine if the responsiveness of normal B cells to LPS(phenol) were due, at least in part, to the stimulation of normal Lyb-5- B cells, the responsiveness of normal neonatal B cells and normal adult B cells that had been pretreated with anti-Lyb-5.1 + C was assessed. It was found that both normal neonatal B cells and normal adult Lyb-5- B cells did respond well to LPS(phenol). Thus, even though LPS(phenol) does not stimulate xid B cells, these data demonstrate that LPS(phenol) is different from other TI-2 antigens. More importantly, these data also demonstrate that xid B cells and normal Lyb-5- B cells are not identical. It is hypothesized that the normal Lyb-5- B cell subpopulation is heterogeneous, consisting of an Lyb-5(1)- and an Lyb-5(2)-B cell subset with the xid mutation blocking the differentiation of Lyb-5(1)-B cells into Lyb-5(2)-B cells.

Animals↗

PubMed1983

[Studies on carcinoembryonic antigen (CEA), lactate dehydrogenase (LDH), and LDH isozymes in the tissue of colorectal carcinoma].

Tissue carcinoembryonic antigen (CEA), lactate dehydrogenase (LDH) and LDH isozymes were measured to assess as tumor markers of colorectal cancer. Tissue samples were collected from carcinoma and the entire layers of normal colorectum at the distance of 1, 2, 4, 6 and 8 cm from the edge of carcinoma. The mean CEA content in cancer tissue was 18143 ng/g and the elevation of CEA content was found in normal tissue within 6 cm from the edge of carcinoma comparing to that in control materials, gradually decreased as the distance from carcinoma increased. LDH activity in cancer tissue was very high and its gradual decrease in normal tissue surrounding tumor with the distance from carcinoma was recognized. LDH isozyme ratio (LDH4 + LDH5/LDH1 + LDH2) and H subunit % (LDH1 + 3/4LDH2 + 1/2LDH5 + 1/4LDH) in cancer tissue were obviously different from those of control materials, LDH isozyme ratio in normal tissue within 4 cm from the edge of carcinoma and H subunit % within 6 cm showed the significant difference to those in control materials. The author concludes that the increase of tissue CEA content, LDH activity and the deviation of LDH isozymatic pattern in normal tissue could be early signs of malignancy before the morphologic changes.

Adenocarcinoma↗

PubMed1983

[Significance of H-Y antigen in Y-linked gene and sex differentiation].

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Animals↗

PubMed1983

Effects of B2-deficiency on lipoperoxide and its scavenging system in the rat lens.

The level of NADPH-cytochrome c reductase, glutathione reductase and hepatic B2 was significantly decreased in rats fed a B2-deficient diet. The synthesis of ester forms of B2 was decreased more in the lenses of rats fed this diet for 8 weeks than in those fed this diet for 4 weeks. The glutathione reductase activity in the lens decreased as the duration of a B2-deficient diet increased; this may be ascribable to a decrease in the formation in the lenticular riboflavin ester as a coenzyme. The level of lipid peroxides increased in the serum and lenses of rats on the B2-deficient diet, however, compared to the controls, the changes in the activities of superoxide dismutase and glutathione S-transferase were not significant. Lenticular glutathione peroxidase activity was decreased significantly in rats fed the B2-deficient diet for 8 weeks. We suggest that B2 deficiency brings about metabolic disorders in the redox system to maintain lens transparency, that lipoperoxide is accumulated, and that the efficiency of the scavenging system and the activities of glutathione peroxidase and reductase are decreased.

Animals↗