A new method of optical detection of yeast acidification power

We describe here a newly developed method for a contact-free optical pH measurement in yeast suspensions supplemented with glucose, and containing the pH sensitive triphenylmethane dye bromocresol green. It is suitable for performing the acidification power test (based on measuring the rate of pH drop of yeast suspension caused by active extrusion of acidity from cells after glucose addition) used for assessing yeast vitality in fermentation industries. Using this methodology we monitored the pH in yeast suspensions in the course of acidification in the pH range of 3.5–5.3. Optical pH measurement allows simultaneous testing of several samples, minimizes the sample volume, simplifies sample handling and reduces the hands-on time in sample processing.     

Factors affecting the outcome of human blastocyst vitrification

This study aimed to test the feasibility of a programme of semen collection and cryopreservation in Griffon vultures. Four wild-caught individuals kept in captivity because of unrecoverable traumas were used. Semen collection attempts were made twice a week during three consecutive reproductive seasons (December – March) using the abdominal massage method. Ejaculation was successfully induced between late January and late February. Semen collection efficiency was rather low (27.9%) and it did not vary among individuals (p > 0.05). No differences were found in ejaculate volumes (12.5 +/- 9.1 μl), spermatozoa concentration (28.4 +/- 30.9 million cells/ml) and viability (61.3 +/- 13.9%) among the 4 vultures. ATP values differed among the four vultures (p < 0.001); B showed higher nucleotide concentration than both C and D, while it did not differ form A, whose values were higher compared with D. After freezing and thawing, semen in vitro viability, DNA integrity and ATP intracellular concentration were determined. Spermatozoa viability after thawing did not differ among the four individuals (52.6 +/- 5.8 in A, 53.4 +/- 4.6 in B, 50.4 +/- 3.2 in C, 42.5 +/- 2.7 in D), but it decreased significantly compared to fresh semen (p < 0.05). During 4 hrs in vitro culture, spermatozoa collected from B maintained over time a higher viability in vitro when compared to A, C and D. As evaluated by the comet assay method, DNA fragmentation after freezing and thawing did not differ in the 4 vultures. ATP concentration in frozen/thawed semen was significantly lower than in fresh semen (p < 0.0001). This study indicates that semen cryopreservation can be considered as a useful tool in the conservation of Griffon vulture genetic resources, but further studies are needed to optimize this technique.     

Factors affecting the outcome of human blastocyst vitrification

With single blastocyst transfer practice becoming more common in ART, there is a greater demand for a convenient and reliable cryostorage of surplus blastocysts. Vitrification has emerged in the last decade as an alternative promising substitute for slow freezing. Blastocysts represent a unique challenge in cryostorage due to their size, multicellular structure and presence of blastocoele. The continuous acquisition of experience and introduction of many different technological developments has led to the improvement of vitrification as a technology and improved the results of its application in blastocyst cryostorage. The current information concerning safety and efficacy of the vitrification of blastocysts will be reviewed along with the variables that can impact the outcome of the procedure.     

Factors affecting the ethanol productivity of yeast in molasses

The ethanol production by a laboratory yeast strain, X2180-1B, was less than half that by an alcohol yeast, YOY655, in a molasses medium containing 30% sugars, although X2180-1B produced approximately the same amount of ethanol as YOY655 in a nutrition medium with the same sugar content. The weak productivity of X2180-1B in the molasses was ascribed to the limitation of sucrose hydrolysis in the molasses. The invertase activity of X2180-1B was 0.019 (mmol sucrose/min/mg protein) in the nutrition medium, but substantially zero in the molasses, while that of YOY655 was 1.75 in the nutrition medium and 1.15 even under the inhibitory conditions in molasses. External addition of invertase greatly enhanced the ethanol productivity of only X2180-1B. The inhibitory factors of invertase in molasses were heat-stable and dialyzable substances.     

Factors affecting the oligomeric structure of yeast external invertase

It has been assumed that yeast external invertase is a dimer, with each subunit composed of a 60-kDa polypeptide chain. We now present evidence that at its optimal pH of 5.0, the predominant form of external invertase is an octamer with an average size of 8 X 10(5) Da. During ultracentrifugation the octamer dissociated to lower molecular weight forms, including a hexamer, tetramer, and dimer. All forms of the enzyme were shown to possess identical specific activities and to contain a similar carbohydrate to protein ratio. Although the monomer subunits (1 X 10(5) Da) were heterogenous in carbohydrate content, each subunit possessed nine oligosaccharide chains. When stained for protein and enzyme activity following sodium dodecyl sulfate-polyacrylamide gel electrophoresis, only the oligomeric form of the enzyme appeared to be active. Thus, on partially inactivating invertase with 4 M guanidine hydrochloride both octamer and monomer were evident on the gels but only the former was active. Similarly, incubating at pH 2.5 in the presence of sodium dodecyl sulfate yielded only inactive monomer. The monomer, unlike the active oligomeric aggregate, was unable to hydrolyze sucrose after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Consistent with the in vitro studies, freshly prepared yeast lysate was shown to contain the octameric species of external invertase as the major active form of this enzyme. From these studies and others which employed deglycosylated invertase, it is concluded that the carbohydrate component of external invertase contributes not only to stabilizing enzyme activity, but also to maintaining its oligomeric structure.     

Factors affecting outcome in free-tissue transfer in the elderly

Free-tissue transfers have become the preferred surgical technique to treat complex reconstructive defects. Because these procedures typically require longer operative times and recovery periods, the applicability of free-flap reconstruction in the elderly continues to require ongoing review. The authors performed a retrospective analysis of 100 patients aged 65 years and older who underwent free-tissue transfers to determine preoperative and intraoperative predictors of surgical complications, medical complications, and reconstructive failures. The parameters studied included patient demographics, past medical history, American Society of Anesthesiology (ASA) status, site and cause of the defect, the free tissue transferred, operative time, and postoperative complications, including free-flap success or failure. The mean age of the patients was 72 years. A total of 46 patients underwent free-tissue transfer after head and neck ablation, 27 underwent lower extremity reconstruction in the setting of peripheral vascular disease, 10 had lower extremity traumatic wounds, nine had breast reconstructions, four had infected wounds, two had chronic wounds, and two underwent transfer for lower extremity tumor ablation. Two patients had an ASA status of 1, 49 patients had a status of 2, 45 patients had a status of 3, and four had a status of 4. A total of 104 flaps were transferred in these 100 patients. There were 49 radial forearm flaps, 34 rectus abdominis flaps, seven latissimus dorsi flaps, seven fibular osteocutaneous flaps, three omental flaps, three jejunal flaps, and one lateral arm flap. Four patients had planned double free flaps for their reconstruction. Mean operative time was 7.8 hours (range, 3.5 to 16.5 hours). The overall flap success rate was 97 percent, and the overall reconstructive success rate was 92 percent. There were six additional reconstructive failures related to flap loss, all of which occurred more than 1 month after surgery. Patients with a higher ASA designation experienced more medical complications (p = 0.03) but not surgical complications. Increased operative time resulted in more surgical complications (p = 0.019). All eight cases of reconstructive failure occurred in patients undergoing limb salvage surgery in the setting of peripheral vascular disease. Free-tissue transfer in the elderly population demonstrates similar success rates to those of the general population. Age alone should not be considered a contraindication or an independent risk factor for free-tissue transfer. ASA status and length of operative time are significant predictors of postoperative medical and surgical morbidity. The higher rate of reconstructive failure in the elderly peripheral vascular disease population compares favorably with other treatment modalities for this disease process.     

Factors affecting soil acidification under legumes I. Effect of potassium supply

This study examined the effect of K (as K₂SO₄) supply on acid production under N₂-fixing plants of lupin (Lupinus angustifolius L. cv. Gungurru) and clover (Trifolium subterraneum L. cv. Dalkeith) grown in a K-deficient soil with a low pH buffer capacity for 55 days in the glasshouse at 20/12 °C (day/night). Increasing K supply up to 240 mg K kg^-1 soil markedly increased plant growth of both species but clover growth was more responsive than lupin. Growing plants for 55 days decreased soil pH by 0.65–0.85 units under lupin and 0.45–0.83 units under clover. The amounts of H⁺ produced per kg biomass (specific acid production) were the highest at the nil K supply, generally decreased with increasing K level up to 30 mg K kg^-1 under lupin and up to 120 mg K kg^-1 soil under clover and only slightly increased with further increasing K under lupin. Increasing K₂SO₄ supply proportionally increased plant uptake of K and SO ₄ ^2- but generally decreased concentrations of Ca, Mg, Na, P and Cl. Specific acid production correlated well with concentrations of excess cations and ash alkalinity, and total acid production was strongly correlated with total excess cations and total ash alkalinity in plants. These relationships were not affected by K treatment and species. Specific acid production also correlated with plant Ca concentration but not with K concentration. In addition, lupin and clover extruded similar amounts of H⁺ per kg biomass produced. It is suggested that application of K₂SO₄ does not have a significant impact on acid production by lupin and clover.     

Factors affecting the performance of crossflow filtration of yeast cell suspension

Factors affecting the performance of crossflow filtration were investigated with a thin-channel module and yeast cells. In crossflow filtration of Saccharomyces cerevisiae cells cultivated with YPD medium (Yeast extract, polypeptone, and dextrose) and suspended in saline, a steady state was attained within several minutes when the cell concentration was low and the circulation flow rate was high. The steady-state flux and the change in flux during the initial unsteady state were explained well by conventional filtration theory, with the amount of cake deposited and the mean specific resistance to the cake measured in a dead-end filtration apparatus used in calculation. When the circulation flow rate was lower than a critical value, a part of the channel of the crossflow filtration module was plugged with cell cake, and thus the steady-state flux was low. In crossflow filtration of suspensions of commercially available baker's yeast, the flux gradually decreased, and the flux after 8 h of filtration was lower than the value calculated by filtration theory. Fine particles contaminating the baker's yeast was responsible for the decrease. A similar phenomenon was responsible for the decrease. A similar phenomenon was observed in crossflow filtration of a broth of S. cerevisiae cells cultivated in molasses medium, which also contains such particles, had no effect of the permeation flux during crossflow filtration. (c) 1993 John Wiley & Sons, Inc.     

Factors affecting the inhibition of yeast plasma membrane ATPase by vanadate

Inhibition of yeast plasma membrane ATPase by vanadate occurs only if either Mg2+ or MgATP2- is bound to the enzyme. The dissociation constant of the complex of vanadate and inhibitory sites is 0.14-0.20 microM in the presence of optimal concentrations of Mg2+ and of the order of 1 microM if the enzyme is saturated with MgATP2-. The dissociation constants of Mg2+ and MgATP2- for the sites involved are 0.4 and 0.62-0.73 mM, respectively, at pH 7. KCl does not increase the affinity of vanadate to the inhibitory sites as was found with (Na+ + K+)-ATPase. On the other hand, the effect of Mg2+ upon vanadate binding is similar to that upon (Na+ + K+)-ATPase, and the corresponding affinity constants of Mg2+ and vanadate for the two enzymes are of the same order of magnitude.     

Factors affecting quality of informed consent.

OBJECTIVE--To examine the factors influencing quality of informed consent. DESIGN--Prospective study comprising interviews with patients and patients'' completing standard questionnaires. SETTING--Academic surgical unit of large teaching hospital. PATIENTS--265 patients undergoing intrathoracic, intraperitoneal, and vascular surgical procedures. Of these patients, 192 have been followed up for six months. MAIN OUTCOME MEASURES--Patients'' recall of information at various points in the study; this score was compared by age, provision of written information, cognitive function, intelligence quotient (IQ), mood state and personality traits, and health locus of control. RESULTS--The patients were best informed immediately after signing the consent form and from then on recall of information deteriorated. A total of 172/250 (69%) patients admitted to not reading the consent form before signing it. Old age adversely affected recall of information at all assessment points. Impaired cognitive function reduced information recall only during the stay in hospital. Patients with above average IQs handled information better than those with a lower IQ except immediately after the signing of consent forms. Patients with an internal locus of health control (that is, those who believed their health to be in their own control) were better informed than those with an external locus of health control. Operation information cards improved recall only on the day of discharge. CONCLUSION--Elderly patients and patients with below average IQ, impaired cognitive functions, and an external locus of control have poor information recall. Written information may be more useful if given before admission to hospital.     

Factors affecting mycelial to yeast phase conversion and growth of the yeast phase ofHistoplasma capsulatum

A procedure for rapid induction of mycelial to yeast phase (M→Y) conversion ofHistoplasma capsulatum has been devised. Exposure of mycelial fragments to low oxidation-reduction (O/R) potentials (+5 to+65 mv) either aerobically (ascorbic acid treatment) or anaerobically (nitrogen atmosphere) for 18 to 24 hours at 37° C resulted in induction of the M→Y conversion process whether or not an organic sulfur source was available. However, aerobic conditions and a suitable organic sulfur source, such as cysteine, cystine or lanthionine were found essential for outgrowth and maintenance in the yeastlike phase.     

Factors affecting the feeding quality of barley for ruminants

Variation in the nutritive value of barley is well documented. Although bulk density is a convenient means of evaluating barley, it does not fully account for differences observed in animal performance from feeding different sources of barley. Recent data indicate that starch and (or) fiber content may be more reliable criteria for evaluating sources of barley. As progress has been made toward higher starch content and plumper barley, greater attention should be given to proper processing of barley before feeding. Improper processing might well negate potential improvements expected from higher-quality barley.     

Factors affecting goat milk production and quality

Differences between production systems based on grazing and browsing vs. use of harvested feedstuffs in confinement largely depend on specific feedstuffs and plants available and being consumed. Low forage nutrient ingestion should have relatively greater impact on tissue mobilization than milk production in early than later periods of lactation, with a transition to proportionally greater change in milk production in late lactation. However, low body condition at kidding would limit tissue energy mobilization and restrict impact of level of nutrient intake to milk yield and, likewise, tissue mobilization would be less with one vs. two or three milkings per day. As lactation advances after freshening, fat and protein levels decrease with increasing milk yield, and when production declines in mid- to late lactation, fat and protein concentrations increase. Milk production generally peaks at a parity of 3 or 4, thereafter declining slowly. Elevated somatic cell count alone in dairy goats is not a valid indication of mammary infection. Extended lactations offer opportunities to minimize or avoid seasonal fluctuations in milk production and lessen production costs. If differences in performance between suckled and machine-milked dairy goats occur, they may be restricted to or of greater magnitude during the suckling period compared with post-weaning, and differences in milk yield will either be absent or less with one kid compared with greater litter sizes. The magnitude of effects of milking frequency on milk yield is less for goats of low vs. high production potential and with low vs. high diet quality. Likewise, the effect of milking frequency is greater in early and mid-lactation when yield is higher than in late lactation, along with a shorter period of peak production with one vs. two daily milkings. Physical form of the diet can affect production and composition of goat milk, although effects appear of smaller magnitude than in dairy cattle. When tissue is mobilized to support milk production in early lactation, levels of C18:0 and C18:1 cis in milk increase and levels of medium-chain fatty acids decline. Effects of elevated levels of dietary fatty acids on specific long-chain fatty acids in milk and milk products vary with the fatty acid profile of fat sources used.     

Factors affecting oxygen-induced changes in the activity of CTP-dependent lipid kinases in yeast

Previous work from this Laboratory (Szkopińska et al., 1988, Arch. Biochem. Biophys., 266, 124-131) indicated that CTP is a phosphate donor for the synthesis of phosphatidic acid and dolichyl phosphates. The elucidation of the role of mitochondrial membranes and mitochondrial proteins (isolation of rho- mutant) as well as specific detergents and sterols has been the aim of this work.     

Factors affecting the susceptibility of sensitive yeast cells to killer toxin K1

Optimum conditions for action of the killer toxin K1 on sensitive strainS. cerevisiae S6 were established. Maximum killing was reached in a very narrow pH range of 4.5–4.6. Maximum susceptibility to toxin was displayed by highly energized fresh cells from the early exponential phase in the presence of an external energy source (at least 200 mmol/L glucose). Further, maintenance of maximum membrane potential was necessary for killer action, as documented by decreasing toxin activity in the presence of increasing concentrations of KCl. The killing was strongly stimulated in the presence of millimolar concentrations of Ca²⁺ and Mg²⁺.     

Factors encoded by and affecting the holding stability of yeast plasmid pSR1

Summary A 2 m DNA-like plasmid, pSR1, isolated from a strain of Zygosaccharomyces rouxii has three coding frames, P, S and R. Insertional inactivation of R completely abolished the intramolecular recombination, and the defect was complemented by an intact R frame on a coexistent plasmid molecule. The P and S regions were also transactive and important, but not essential, for the stable maintenance of the plasmid molecules. Insertional disruption of the P frame suggested that it produces a protein factor. Similar insertional disruption of the S frame affected the plasmid stability in Z. rouxii and Saccharomyces cerevisiae hosts differently, depending on whether the inserted DNA fragment was a short 8 bp SalI linker or a long (2.2 kb) DNA fragment. Results strongly suggested that the S region encodes two factors, one RNA and the other a protein, and that the S protein is compatible with a sprecific hostfactor in Z. rouxii, but not in S. cerevisiae. In addition, a cis-acting locus, Z, was found at a site in the plasmid molecule where no distinct open reading frames were located. No long direct repeats or inverted repeats were observed in the Z region, such as are found in the REP3 locus of 2 m DNA.