Slower growth of S. typhi in hypertonic media, reported previously by the authors, was contradictory to other workers', results which showed better growth of some species of bacteria. To evaluate furthur the effect of hypertonic sucrose on the growth of S. typhi, organisms were suspended in saline or in blood with or without sodium polyanethol sulfonate (SPS) and stored up to 24 hours. And then viable counts were determined on tryptic soy agar (TSA) and experimental blood cultures were done in tryptic soy broth (TSB) and in TSB with 10% sucrose (TSB-H). S. typhi, suspended in blood and kept for 24 hours, were inoculated into TSB and TSB-H and after 4 hour incubation viable counts were made on TSA and on TSA with 10% sucrose (TSA-H). In this study it was found that, during the 24 hour storage, the viable counts of S. typhi suspended in saline with or without SPS were similar and those suspended in blood with SPS were incereasing. Comparison of the growth in TSB and in TSB-H did not show hyperonic media was better for the cultivation of S. thphi which was kept up to 24 hours before inoculation. On the contrary the growth was slower. Viable counts made on TSA and on TSA-H from the TSB and TSB-H, which were inoculated with S. typhi suspended in blood and incubated for 4 hours, showed similar results indicating TSB-H did not support faster growth. From the results of this experiment and of the previous clinical blood cultures, it is concluded that 0.1% SPS does not give adverse effect on S. typhi during the 24 hour storage and that hypertonic sucrose does not give better result in the cultivation of S. typhi.
Blood/microbiology* ; Culture Media ; Hypertonic Solutions ; Salmonella typhi/drug effects ; Salmonella typhi/growth & development* ; Sucrose/pharmacology*

OBJECTIVETo study the effects of different factors on buds and microtuber. These factors included plant growth substances and sucrose.

METHODstems were selected as explants. The effects of three kinds of factors were studied by orthogonal design method including sucrose, 6-BA, NAA on the buds and microtuber induction. The data were analyzed with range analysis and vadance analysis. RESULT AND CONDUSION: The optimal media to induce many buds from stems were MS + 6-BA 1 mg x L(-1) + NAA 1 mg x L(-1) + sucrose 3%, the effect of the three factors was in sequence of sucrose >6-BA > NAA. The optimal media to induce microtuber from stems were MS+6-BA 1.5 mg x V1 +NAA 1.5 mg x L(-1) + sucrose 5%, the effect of the three factors was in sequence of sucrose >6-BA > NAA.

Culture Media ; Dioscorea ; drug effects ; growth & development ; Plant Growth Regulators ; pharmacology ; Sucrose ; pharmacology ; Tissue Culture Techniques ; methods

Taxus suspension cell culture has the potential to provide a sustainable source of anticancer drug paclitaxel (Taxol) and other taxoids. In the cell culture of Taxus chinensis, Taxuyunnanine C (Tc) is the primary taxoid. To design a rational strategy for redirecting the precursor fluxes from other taxoids into paclitaxel production, we employed Real-time Quantitative PCR (RQ-PCR) to understand the dynamic profiling of key biosynthetic pathway genes of palcitaxel and taxoids during the culture process. Six genes (TASY, TDAT, T5alphaH, TalphaH, T10betaH and T14betaH) were quantified under the process condition of double elicitation by 2,3-dihydroxylpropanyl jasmonate (DHPJA) (100 micromol/L on day 7 and day 12), and sucrose feeding (20 g/L) on day 7. This process treatment led to a high accumulation of Tc at (554.46 +/- 21.28) mg/L 8 days after the first elicitation. Then 9 days after the second elicitation, Tc production was as high as (997.72 +/- 1.51) mg/L. The early pathway genes TASY and TDAT were significantly up-regulated by 182-fold and 98-fold, respectively for the first DHPJA elicitation and by 208-fold and 131-fold, respectively for the second elicitation. The induction occurred after each elicitation lasted for about 24 h before their abundances decreased. Things are somewhat different in the case of the other four genes T5alphaH, TalphaH, T10betaH and T14betaH. For gene TalphaH, it was highly up-regulated by 3061-fold for the first DHPJA elicitation and by 1016-fold for the second elicitation. For the other three genes T5alphaH, T10betaH, T14betaH, they were up-regulated by 13-fold, 38-fold and 20-fold, respectively for the first DHPJA elicitation and by 7-fold, 16-fold and 6-fold, respectively for the second elicitation. The RQ-PCR results showed that there is tight correlation between gene expression and Tc accumulation. Gene expression was in accordance with Tc yield. Elicitation could improve expression of six genes. While along with culture course, high expression of the genes weakened. Elicitation for the second time would promote high expression of the genes again.
Cell Culture Techniques ; Culture Media ; Cyclopentanes ; pharmacology ; Gene Expression Regulation, Plant ; Oxylipins ; pharmacology ; Plant Growth Regulators ; pharmacology ; Sucrose ; pharmacology ; Taxoids ; metabolism ; Taxus ; genetics ; metabolism ; Transcriptome

OBJECTIVETo study the conditions on separation and regeneration of protoplast from Phellinus igniarius.

METHODThe effects of enzymolysis conditions of P. igniarius mycelia on yield of protoplast and culturing conditons on regeneration ratio of protoplast were investigated.

RESULTWhen the 8 days-old mycelia was hydrolysed by 1.5% of lywallzyme adding to driselase of 0. 5% and at 30 degrees C for 3 h and enzymolysis was stablized by sucrose as a stablisher of osmotic pressure, higher yield of P. igniarius protoplast was obtained. If 10 days-old mycelia was used as raw material of enzymolysis and manntol was selected as stablisher of osmotic pressure of enzymolysis, higher regeneration ratio of P. igniarius protoplast also would be obtained in following regeneration step at same time keeping higher yield. For the regeneration processing, it was beneficial for the regeneration of P. igniarius protoplast that PDA plusing mulberry ramulus was used as the culture medium of regeneration and manntol was selected as the osmotic pressure establisher of regeneration culture medium.

CONCLUSIONThe method and conditions to keep both higher yield and regeneration ratio of P. igniarius protoplast were obtained.

Culture Media ; pharmacology ; Fungal Proteins ; pharmacology ; Glucan Endo-1,3-beta-D-Glucosidase ; pharmacology ; Glycoside Hydrolases ; pharmacology ; Mannitol ; pharmacology ; Multienzyme Complexes ; pharmacology ; Osmotic Pressure ; Peptide Hydrolases ; pharmacology ; Polyporaceae ; drug effects ; physiology ; Protoplasts ; drug effects ; physiology ; Regeneration ; drug effects ; Sucrose ; pharmacology ; Temperature

Although ethylene glycol (EG) has been widely used for embryo cryopreservation in domestic animals, few attempts were made to use this molecule to freeze mouse and human embryos. In the few studies that used EG for slow-freezing of mouse and human embryos, complicated protocols for human embryos were used, and the protocols need to be simplified. Besides, freezing mouse morula with EG as a cryoprotectant has not been reported. In this paper, we studied the effects of embryo stages, EG concentration, duration and procedure of equilibration, sucrose supplementation and EG removal after thawing on the development of thawed mouse embryos, using the simple freezing and thawing procedures for bovine embryos. The blastulation and hatching rates (81.92% +/- 2.24% and 68.56% +/- 2.43%, respectively) of the thawed late compact morulae were significantly (P < 0.05) higher than those of embryos frozen-thawed at other stages. When mouse late compact morulae were frozen with different concentrations of EG, the highest rates of blastocyst formation and hatching were obtained with 1.8mol/L EG. The blastulation rate was significantly higher when late morulae were equilibrated in 1.8 mol/L EG for 10 min prior to freezing than when they were equilibrated for 30 min, and the hatching rate of embryos exposed to EG for 10 min was significantly higher than that of embryos exposed for 20 and 30 min. Both rates of blastocyst formation and hatching obtained with two-step equilibration were higher (P < 0.05) than with one-step equilibration in 1.8 mol/L EG. Addition of sucrose to the EG-based solution had no beneficial effects. On the contrary, an increased sucrose level (0.4 mol/L) in the solution impaired the development of the frozen-thawed embryos. In contrast, addition of 0.1 mol/L sucrose to the propylene glycol (PG)-based solution significantly improved the development of the frozen-thawed embryos. Elimination of the cryoprotectant after thawing did not improve the development of the thawed embryos. The cell numbers were less (P < 0.05) in blastocysts developed from the thawed morulae than in the in vivo derived ones. In summary, embryo stage, EG concentration, duration and procedure of equilibration and sucrose supplementation had marked effects on development of the thawed mouse embryos, and a protocol for cryopreservation of mouse embryos is recommended in which the late morulae are frozen in 1.8 mol/L EG using the simple freezing and thawing procedures of bovine embryos after a two-step equilibration and the embryos can be cultured or transferred without EG removal after thawing.
Animals ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Dose-Response Relationship, Drug ; Embryo, Mammalian ; drug effects ; physiology ; Embryonic Development ; physiology ; Ethylene Glycol ; pharmacology ; Female ; Mice ; Morula ; physiology ; Pregnancy ; Sucrose ; pharmacology

OBJECTIVETo compare the effects of the cryoprotectant containing glucose and that containing sucrose on the motility of post-thaw human sperm.

METHODSThe cryoprotectant containing glucose and that containing sucrose were applied to 50 semen samples and the motility of the post-thaw human sperm was compared before and after cryopreservation and between the study groups.

RESULTSThe forward motility and total motility of the sperm were (58.4 +/- 5.7)% and (63.4 +/- 6.1)% before cryopreservation, (43.8 +/- 7.6)% and (48.4 +/- 7.6)% after thawing with the cryoprotectant containing glucose, and(42.6 +/- 8.9)% and (48.0 +/- 8.5)% after thawing with the cryoprotectant containing sucrose. Decreased sperm motility was observed after cryopreservation, with statistic significance (P < 0.01). There was no significant difference in the forward and total motility of the post-thaw sperm between the two cryoprotectants.

CONCLUSIONCryopreservation inflicts obvious damage on sperm. Sucrose is a feasible sperm cryoprotectant.

Adult ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Glucose ; pharmacology ; Humans ; Male ; Semen Preservation ; methods ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects ; physiology ; Sucrose ; pharmacology

OBJECTIVETo study the effect of different factors on induction of microtubers in Zingiber officinale. These factors included NAA, PP333, 6-BA and sucrose.

METHODOrthogonal design and plant tissue culture technique were used.

RESULT AND CONCLUSIONSucrose was the most important factor on the induction of microtubers, followed by PP333 and NAA. 6-BA was the factor which restrained the formation of microtubers. The optimal media to induce microtubers was MS + NAA 1.0 mg x L(-1) + PP333 0.2 mg x L(-1) + sucrose 8%.

Culture Media ; pharmacology ; Ginger ; growth & development ; Plant Growth Regulators ; pharmacology ; Plants, Medicinal ; growth & development ; Rhizome ; drug effects ; growth & development ; Sucrose ; pharmacology ; Tissue Culture Techniques ; methods

Osmotically stabilized media have been reported to increase the recovery rate of various bacteria from blood. This study was made to determine the effect of high concentrations of sucrose on the cultivation of S. typhi from blood. Sucrose in 15% or 30% concentration in the blood culture media retarded the growth. The mean incubation time for the appearance of growth was significantly longer in the media with sucrose. In those blood specimens which rendered growth of S. typhi in both media with and without sucrose, the incubation times were compared; and it was found that the majority of the specimens showed faster growth in the media without sucrose. Experimental cultures showed that the higher the sucrose concentration the lighter and slower were the growths of S. typhi. These tendencies were also observed in the growth of E. coli, P. aeruginosa, S. aureus, beta-hemolytic Streptococcus, alpha-hemolytic Streptococcus and S. pneumoniae.
Culture Media ; Human ; Salmonella typhi/drug effects ; Salmonella typhi/growth & development* ; Salmonella typhi/isolation & purification ; Sucrose/pharmacology*

OBJECTIVETo explore the feasibility of ultra-rapid freezing of human spermatozoa in the cryogenic vial with different concentrations of sucrose solution.

METHODSWe divided 40 normal semen samples prepared with the routine swim-up technique into 6 aliquots, 1 as the control and the other 5 cryopreserved with sucrose solution at the concentrations of 0.15, 0.20, 0.25 and 0.30 mol/L, respectively. After thawing, we determined and compared the motility, progressive motility and plasma membrane integrity of the sperm among the 6 groups.

RESULTSThe motility, progressive motility and plasma membrane integrity of the sperm were significantly lower after thawing than before cryopreservation ([96.2 +/- 1.8]%, [93.8 +/- 2.8]% and [99.0 +/- 0.8 ]%) (P<0.05). Post-thawing sperm motility was (55.5 +/- 6.3)% in the 0.20 mol/L sucrose group, significantly higher than in the 0.15, 0.25 and 0.30 mol/L groups ([45.9 +/- 6.6]%, [50.4 +/- 9.4]% and [45.5 +/- 11.2]%) (P<0.05), and it was (53.6 +/- 5.0)% in the conventional freezing group, with no statistically significant difference from the 0.20 and 0.25 mol/L sucrose cryopreservation groups (P> 0.05), but remarkably higher than in the 0.15 and 0.30 mol/L groups (P<0.05). Post-thawing progressive sperm motility exhibited no statistically significant differences between the 0.20 mol/L sucrose and conventional freezing groups ([44.4 +/- 7.4]% vs [42.3 +/- 8.1]%, P>0.05), but markedly higher in both than in the 0.15, 0.25 and 0.30 mol/L sucrose groups ([37.1 +/- 8.3 ]%, [33.1 +/- 9.2]% and [22.0 +/- 9.1]%) (P<0.05). Post-thawing plasma membrane integrity was significantly higher in the 0.20 mol/L sucrose cryopreservation group ( [70.1 +/- 6.9]%) than in either the conventional freezing group ([63.1 +/- 6.8]%) or the 0.15, 0.25 and 0.30 mol/L sucrose groups ([57.7 +/- 8.3]%, [63.5 +/- 10.7]% and [57.8 +/- 12.9]%) (P<0.05).

CONCLUSIONAs a simple, safe and effective method, ultra-rapid freezing with sucrose solution at the final concentration of 0.20 mol/L can be used for the cryopreservation of human spermatozoa.

Cell Membrane ; drug effects ; Cryopreservation ; methods ; Humans ; Male ; Semen Preservation ; methods ; Sperm Motility ; drug effects ; Sucrose ; administration & dosage ; pharmacology

Effects of sucrose concentrations and light on the growth and production of total isoflavones and puerarin in Pueraria phaseoloides hairy roots cultured onto solid MS media supplemented with 1%, 3%, 5%, 7% and 9% sucrose, respectively, were investigated. The results showed that among the sucrose concentrations tested, 3% sucrose in the medium enhanced the growth and stimulated accumulation of total isoflavones and puerarin in P. phaseoloides hairy roots, After cultured for 20 days, the biomass of hairy roots reached 0.48 g (DW dry weight)/flask and its contents of total isoflavones and puerarin were 25.44 mg/g (DW) and 11.64 mg/g (DW), respectively. In comparison with 3% sucrose, the dry weight proliferation of hairy roots cultured with 5% sucrose was increased by 7.0%, while cultured with 1%, 7% and 9% sucrose, the dry weight proliferation of hairy roots was decreased by 62.4%, 42.8% and 65.3%, their total isoflavones content was decreased by 57.4%, 13% and 33.4% and their puerarin content was decreased by 47.9%, 15.8% and 35.1%; but their content of total soluble sugars was increased 0.52, 1.45 and 1.54 times, respectively. Compared with hairy roots in blue light and white light, the biomass of hairy roots cultured in the dark for 30 days was 0.83 g (DW)/flask and was increased by 37.1% and 23.3%, respectively. The content of total isoflavones in hairy roots cultured in white light was as much as 1.15 times and 1.19 times that in blue light and in the dark, respectively. It was also observed that hairy roots cultured in blue light and white light partly became light green and that blue light could inhibit accumulation of puerarin in hairy roots and the puerarin content in hairy root cultured in white light and in the dark were 1.61 times and 1.52 times that in blue light, respectively.
Culture Media ; Isoflavones ; biosynthesis ; Light ; Plant Roots ; growth & development ; metabolism ; Pueraria ; growth & development ; metabolism ; Sucrose ; pharmacology ; Tissue Culture Techniques ; methods