Effects of culture medium composition and PEG on hyperhydricity in Dendrobium officinale

Hyperhydricity is a physiological disorder during plant tissue culture that seriously affects regeneration and micropropagation. In this study, Dendrobium officinale plantlets were cultured on solid Murashige and Skoog (MS) medium supplemented with plant growth regulators and various concentrations of sucrose, agar, and polyethylene glycol (PEG)-6000 to explore the effect of osmotic stress on hyperhydricity. The results show that low concentrations of sucrose or agar, as well as PEG-6000 at various concentrations, significantly increase the hyperhydric rate of D. officinale, whereas high concentrations of sucrose or agar did not. Furthermore, high concentrations of PEG-6000 significantly increase total water content, free-water content, relative electrical conductivity, and peroxidase (POD) activity of D. officinale plantlets, whereas they significantly decrease bound-water content, proline content, soluble protein content, soluble sugar content, and superoxide dismutase (SOD) activity. These results indicate that PEG-6000 disrupts the antioxidant system and water metabolism in D. officinale plantlets, as well as increases cell membrane permeability, which might be the key factors for the occurrence of hyperhydricity in this species.

     

Evaluation of a new presumptive medium for group D streptococci.

A new medium designated as D streptococcus-enterococcus broth was formulated and evaluated for the enrichment and isolation of strains of serological group D streptococci. This medium was made by modifying Todd-Hewitt broth. Most-probable-number multiple-tube and membrane filter techniques were employed to estimate the numbers of enterococci in known cultures, wastewater, and other samples. Preliminary most-probable-number counts with this medium were as much as 3 logs higher than those counts obtained from four other media with which it was compared. The methodology for using this medium to estimate the numbers of group D streptococci in water is discussed.     

Evaluation of a new presumptive medium for group D streptococci.

A new medium designated as D streptococcus-enterococcus broth was formulated and evaluated for the enrichment and isolation of strains of serological group D streptococci. This medium was made by modifying Todd-Hewitt broth. Most-probable-number multiple-tube and membrane filter techniques were employed to estimate the numbers of enterococci in known cultures, wastewater, and other samples. Preliminary most-probable-number counts with this medium were as much as 3 logs higher than those counts obtained from four other media with which it was compared. The methodology for using this medium to estimate the numbers of group D streptococci in water is discussed.     

Effects of medium composition and culture duration on in vitro morphogenesis of sweet potato

In vitro morphogenesis of sweet potato (Ipomoea batatas) shoot explants after cultures in callus initiation medium (CIM) with two sucrose contents and plant regeneration medium (PRM) with three growth regulator combinations for different durations was studied. After 4 weeks, explants on 5 % sucrose CIM had significantly more shoots but similar or lower root fresh mass and callus fresh mass than those on 3 % sucrose CIM subsequent to transfer for 6 weeks on all three PRM. Cultures transferred to growth regulator-free PRM after 4 and 12 weeks on 5 % sucrose CIM formed plants through organogenesis and embryogenesis, respectively. Embryogenic cultures from 4 weeks on CIM + 10 weeks on callus proliferation medium when transferred to PRM without growth regulator for 4 and 8 weeks produced multiple embryos in the prior and both embryos and shoot buds in the later.     

Gentamicin-based medium for the isolation of group D streptococci and application of the medium to water analysis.

Gentamicin-thallous-carbonate (GTC) medium contained (per liter): 40.0 g of Trypticase soy agar, 5.0 g of KH(2)PO(4), 2.0 g of NaHCO(2), 1.0 g of glucose, 1.0 g of esculin, 0.5 g of thallous acetate (TA), 0.5 g of ferric citrate, 0.75 ml of Tween 80, and 2.5 mg of gentamicin sulfate. The NaHCO(3) (20 ml of a 10% solution that had been heated to boiling) was added after sterilization of the basal medium. The spread plate technique was used to compare GTC agar with Pfizer selective enterococcus, TA, and KF agars by using sewage as well as bovine and swine fecal samples. Significantly greater numbers of group D streptococci were recovered on GTC agar than on Pfizer selective enterococcus or KF agars, within and over all samples. Higher counts also were obtained on GTC than on TA agar, but the differences were not statistically significant. The percentage of false positives was about the same for all four media. Samples of riverwater also were plated on GTC, TA, and KF agars, and significantly higher recoveries were obtained with GTC agar. GTC agar was superior to the other media examined primarily because of increased recoveries of Streptococcus bovis and S. equinus; other advantages of GTC agar were large colony size and short (24-h) incubation period. The percentage of false positives from riverwater was 13% for GTC agar and 0% for TA and KF agars; therefore, confirmation would be necessary when GTC agar is used with some types of environmental samples.     

Interaction of group A streptococci with a culture of HEp-2 cells]

An initial stage of the interaction of the virulent (matt) and avirulent (glossy) strains of group A streptococcus with the human epithelial tissue (Hep-2) was studied. M+ and M- variants of three strains of group A streptococcus belonging to serological types 2 and 4 possessed different biological activity against the Hep-2 epithelial cells in the monolayer. M+-variant actively affected the great majority of the cells of the monolayer, multiplied of their surface and formed microcolonies. M--variant affected only an insignificant number of cells and failed to multiply on them. In difference from M+-streptococci, the activity of M+-variants is explained by their capacity to adhere to the surface of the animal cells irreversibly. This process started immediately and terminated by 1 1/2 hours of the microbial incubation in vitro with the Hep-2 monolayer. Trypsin treatment of M+-streptococci sharply diminished their capacity to adhesion, this apparently being the result of M-protein digestion. The data presented point to the important role played by M-protein in the phenomenon of the streptococci adhesion to the epithelial cells.     

The modification of the cell wall proteins in group A streptococci type M 29 under the influence of spermidine contained in the culture medium

The addition of spermidine into growth medium used for the cultivation of group A streptococci, type M 29, leads to changes in the amino acid composition of cell walls and surface proteins isolated by the method of E. H. Beachey et al. The separation of surface proteins into fibrinogen-binding proteins and fibrinogen receptors by affinity chromatography techniques on cellulose with covalently bound fibrinogen indicates that the proportion of these proteins in pepsin extracts obtained from different strains varies. Both spermidine and avirulent strains have similar content of fibrinogen-binding proteins, although these proteins are absent in virulent strains. Different amounts of fibrinogen receptors are extracted from all strains. As shown in the enzyme immunoassay, fibrinogen receptors contain no group-specific polysaccharide A, Fc-receptors and interact with total antiserum to group A streptococci, type M 29 [correction of 28]. Fibrinogen receptors isolated from the strains under study have been found to have similar amino acid composition. On the basis of these results we believe that neither receptor capacity to fibrinogen nor amino acid composition is indicative of the protective properties of protein M.     

Effect of medium composition on the susceptibility of oral streptococci to mercuric chloride

Although there is considerable interest in identifying mercury-resistant bacteria, no standardized assay exists for this purpose. In this study, the effect of the composition of the medium on the susceptibility of oral streptococci to HgCl₂ was investigated. The minimum inhibitory concentration (MIC) of HgCl₂ for 52 streptococcal strains and the reproducibility of MIC values for Hg-sensitive and Hg-resistant strains was determined with 11 different media. Addition of blood increased the MIC values, and some media (tryptone soya agar, with or without blood) could not discriminate between Hg-sensitive and Hg-resistant strains. The proportion of streptococci that appeared to be resistant to Hg was very high (>70%) on some media (mitis-salivarius, tryptone soya, Columbia), but not on others (Mueller-Hinton, Brain Heart Infusion, Isosensitest). The MICs of the control strains varied considerably on different testing occasions for tryptone soya agar (with and without blood), Isosensitest agar, and Columbia agar (with blood). Mueller-Hinton (without blood) appeared to be the most suitable medium for isolating Hg-resistant oral streptococci. Received: 21 December 2001 / Accepted: 17 January 2002     

Effect of culture medium composition on callus formation and regeneration in oil flax anther culture

Regeneration ability in callus cultures from anthers of two hybrid genotypes of oil flax was studied on N6 and LMA-1 nutrient media at various concentrations of cytokinine 6-benzylamynopurine (BAP). It was shown that callus grew and developed better at BAP concentrations of 2 mg/l, comparing with 4 and 6 mg/l. Shoot and root regeneration was observed in F1 genotype 6-8-gnezdny x M22 only and did not depend on BAP concentration in the medium and on the medium composition itself. Transfer onto fresh medium often stimulated dedifferentiation of the regenerated structures.     

Tetracycline resistance in pneumococci and group A streptococci. Report of an ad-hoc study group on antibiotic resistance.

In a nationwide survey in the spring of 1975 into the prevalence of tetracycline resistance among pneumoccoci and group A streptococci isolates, 21 laboratories reported the sensitivity of isolates and details of the patients. Altogether 13% of the 1528 pneumococci isolated were resistant to tetracycline, but there were wide geographical variations. Thirty-six per cent of the 1515 streptococci isolated were resistant, and again there was considerable geographical variation. A high level of resistance in one organism did not correlate with a high level in the other. For both organisms resistance was commoner among inpatients and those aged 50 or over. Tetracycline should probably not be the drug of choice in penicillin-sensitive patients with group A streptococcal infections, but geographical variations were so wide that decisions on treatment are best made on the basis of local survey data.     

Effect of genotype and medium composition on flax Linum usitatissimum L. anther culture

Identification of responsive genotypes and development of efficient protocols are the prerequisite to an effective doubled haploid production system in applied breeding programs. Evaluation of 16 low linolenic flax (Linola) genotypes/populations with diverse genetic backgrounds from a Linola breeding program using A₂₂C medium containing 9% sucrose (A₂₂C-9) led to the identification of a number of responsive genotypes. For 96-3-F₁ hybrid, callus induction was greater in modified NLN medium containing 12% sucrose (NLN-12) than in A₂₂C-9. But there was no difference in shoot regeneration between NLN-12 and A₂₂C-9. For 96-45-F₁ hybrid, there was no difference in callus production between the two media. However, A₂₂C-9 had a greater shoot regeneration than NLN-12. In comparison to sucrose, lactose was found to increase callus induction from anthers for all three genotypes tested. However, the effect of lactose on shoot regeneration appeared to be genotype-dependent.     

Antibiotic sensitivity of beta-hemolytic Streptococcus group A on a new nutrient medium for the cultivation of streptococci

Sensitivity of 167 strains of beta-hemolytic streptococci of group A was studied with the method of serial dilutions on a solid agar medium for cultivation of streptococci. The medium was developed at the I. I. Mechnikov Research Institute of Vaccines and Sera. It does not require addition of blood or serum. The strains were found to be highly sensitive to penicillin, cephalothin and erythromycin. The number of the strains resistant to tetracycline, streptomycin, gentamycin, levomycetin (chloramphenicol) and ristomycin amounted to 51, 36, 23, 1.8 and 1.8 per cent, respectively. One of the strains (0.6 per cent) was resistant to lincomycin. Strains with multiple resistance were isolated. The necessity of regular control of distribution of antibiotic resistance among staphylococci is indicated.     

Dynamics of proteins in a culture broth growing beta-hemolytic streptococci group C

The dynamic study of the protein spectrum of culture fluid during the growth of beta-hemolytic streptococcal strain H46A has been carried out by the methods of electrophoresis and isoelectrofocusing in polyacrylamide gel. Changes in the protein spectrum have phasic character and, on the whole, reflect the state of the microbial population, the presence of fractions corresponding to streptokinase and streptolysins being detected at all phases of growth. The electrophoretic mobility of streptokinase perceptibly changes at the end of the logarithmic phase; as shown by electrofocusing, at all stages of the population growth the heterogeneity of streptokinase is observed.     

培养基质量对脑膜炎奈瑟菌培养结果的影响

通过对A群C群流脑多糖疫苗生产用培养基质量指标与培养脑膜炎奈瑟菌菌数之间的关系进行分析,发现不同型别的脑膜炎奈瑟菌在相同的培养基和培养条件下具有不同的培养结果,对培养基总氮含量、氨氮含量、氯化钠含量进行检测分析,在一定范围内并未发现与培养菌数之间明显的关系,但这些质量指标作为培养基制备过程的质量控制,保证不同批次培养基始终保持相对稳定、可控的质量,具有十分重要的意义。     

Effect of group racial composition on weight loss in African Americans

Objective: We do not know how racial composition of a group influences behavior change for African Americans (AAs) in group‐based weight loss programs. We tested the hypothesis that AA who participate in all AA weight loss intervention groups will lose more weight than AA who participate in mixed race groups. Methods and Procedures: This observational study was ancillary to Phase 1 of the Weight Loss Maintenance Study, a multi‐center trial of strategies to maintain weight loss after a 20‐week behavior modification program. Three of four centers recruited several all‐AA intervention groups. Remaining groups were combinations of AA and non‐AA participants. All participants received the same weight loss intervention. Change in weight was the primary outcome, comparing participants of all‐AA groups with AA participants of mixed race groups conducted by the same AA interventionists. Secondary outcomes included measures of intervention adherence and behavior change. Results: Participants of all‐AA groups (n = 271) were comparable to other AA participants (n = 106). The mean proportion of AA in mixed race groups was 56%. All‐AA group participants had similar weight loss as those in mixed groups (?4.2 vs. ?4.2 kg, P = 0.97). There were no differences between the groups in mean number of sessions attended or changes in dietary intake. Discussion: Significant weight loss was observed in both groups, with no effect of group composition on adherence or weight loss outcomes. Special logistics to accommodate all‐AA groups may not be necessary. Despite varying instructional environments, AA appeared to respond positively to intervention messages with significant changes in dietary intake, physical activity (PA), and weight.