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[The influence of different cryoprotectant buffers on the fertilizing ability of human sperm "in vitro". I. The choice of cryoprotective buffers and methods of freezing].

We attempted to find the optimal way of cryopreservation of male gametes. We have been tested 9 cryoprotectants currently existing in the world literature. These fluids have been so far separately investigated in different centers of infertility treatment. In this study we compared the effectivity of these fluids in preservation of viable sperm. Basic seminological parameters and two variants of freezing were tested. There were noted significant differences between fast and slow cooling of sperm samples. Nevertheless after fast cooling we found a marked decrease of seminological parameters in preserved sperm when compared to fresh samples.

Cryopreservation↗

Effect of pH buffer molecules on the light-induced currents from oriented purple membrane.

The effect of pH buffers on the microsecond photocurrent component, B2, of oriented purple membranes has been studied. We found that under low salt conditions (less than 10 mM monovalent cationic salt) pH buffers can dramatically alter the waveform of the B2 component. The effect is induced by the protonation process of the buffer molecules by protons expelled from the membrane. These effects can be classified according to the charge transition upon protonation of the buffer. Buffers that carry two positive charges in their protonated form add a negative current component (N component) to B2. Almost all of the other buffers add a positive current component (P component) to B2, which is essentially a mirror image of the N component. Buffers with a pK less than 5.5 have only a small positive buffer component. The pH dependence of the buffer effect is closely related to the pK of the buffer; it requires that the buffer be in its unprotonated form. The rise time of the buffer component increases with the concentration of the buffer molecules. All the buffer effects can be inhibited by the addition of 5 mM of a divalent cation such as Ca2+. Reducing the surface potential slows down the N component but accelerates the P component without affecting the amplitude of the buffer effect significantly. Many of the buffer effects can be explained if we assume that upon protonation of the buffer by a proton expelled from the membrane by light, the buffer molecules move toward the membrane. This backward movement of buffer molecules forms a counter current very similar to that due to cations discussed in Liu, S. Y., R. Govindjee, and T. G. Ebrey. (1990. Biophys. J. 57:951-963).

Bacteriorhodopsins↗

The quantitation of buffering action I. A formal & general approach.

BACKGROUND: Although "buffering" as a homeostatic mechanism is a universal phenomenon, the quantitation of buffering action remains controversial and problematic. Major shortcomings are: lack of a buffering strength unit for some buffering phenomena, multiple and mutually incommensurable units for others, and lack of a genuine ratio scale for buffering strength. Here, I present a concept of buffering that overcomes these shortcomings. THEORY: Briefly, when, for instance, some "free" H+ ions are added to a solution (e.g. in the form of strong acid), buffering is said to be present when not all H+ ions remain "free" (i.e., bound to H2O), but some become "bound" (i.e., bound to molecules other than H2O). The greater the number of H+ ions that become "bound" in this process, the greater the buffering action. This number can be expressed in two ways: 1) With respect to the number of total free ions added as "buffering coefficient b", defined in differential form as b = d(bound)/d(total). This measure expresses buffering action from nil to complete by a dimensionless number between 0 and 1, analogous to probabilites. 2) With respect to the complementary number of added ions that remain free as "buffering ratio B", defined as the differential B = d(bound)/d(free). The buffering ratio B provides an absolute ratio scale, where buffering action from nil to perfect corresponds to dimensionless numbers between 0 and infinity, and where equal differences of buffering action result in equal intervals on the scale. Formulated in purely mathematical, axiomatic form, the concept reveals striking overlap with the mathematical concept of probability. However, the concept also allows one to devise simple physical models capable of visualizing buffered systems and their behavior in an exact yet intuitive way. CONCLUSION: These two measures of buffering action can be generalized easily to any arbitrary quantity that partitions into two compartments or states, and are thus suited to serve as standard units for buffering action. Some exemplary treatments of classical and non-classical buffering phenomena are presented in the accompanying paper.

Buffers↗

Effects of buffered saline solution on nasal mucociliary clearance and nasal airway patency.

OBJECTIVE: To compare the effects of buffered hypertonic and buffered normal saline nasal spray on mucociliary clearance and nasal airway patency. STUDY DESIGN AND SETTING: Double-blind trial with subjects acting as their own controls. Tertiary care academic medical center. RESULTS: Buffered hypertonic saline and buffered normal saline both improved saccharine clearance times ( P < 0.0001 for buffered hypertonic and P = 0.002 for buffered normal saline). Buffered hypertonic saline improved saccharine clearance times more than buffered normal saline (39.6% vs 24.1%, P = 0.007). Neither buffered hypertonic nor buffered normal saline significantly affected nasal airway patency. CONCLUSIONS: Both buffered hypertonic and buffered normal saline nasal spray significantly improved saccharine clearance times without affecting nasal airway patency. Buffered hypertonic saline affected saccharine clearance times to a greater degree than buffered normal saline. CLINICAL SIGNIFICANCE: Buffered hypertonic and buffered normal saline sprays both improve mucociliary clearance and should therefore be beneficial in conditions such as rhinitis and sinusitis, which are associated with disruption of mucociliary clearance. However, these sprays do not appear to affect the nasal airway. Patients may therefore benefit from other treatments for "nasal congestion." EBM RATING: B-2.

Administration, Intranasal↗

The quantitation of buffering action II. Applications of the formal & general approach.

BACKGROUND: The paradigm of "buffering" originated in acid-base physiology, but was subsequently extended to other fields and is now used for a wide and diverse set of phenomena. In the preceding article, we have presented a formal and general approach to the quantitation of buffering action. Here, we use that buffering concept for a systematic treatment of selected classical and other buffering phenomena. RESULTS: H+ buffering by weak acids and "self-buffering" in pure water represent "conservative buffered systems" whose analysis reveals buffering properties that contrast in important aspects from classical textbook descriptions. The buffering of organ perfusion in the face of variable perfusion pressure (also termed "autoregulation") can be treated in terms of "non-conservative buffered systems", the general form of the concept. For the analysis of cytoplasmic Ca++ concentration transients (also termed "muffling"), we develop a related unit that is able to faithfully reflect the time-dependent quantitative aspect of buffering during the pre-steady state period. Steady-state buffering is shown to represent the limiting case of time-dependent muffling, namely for infinitely long time intervals and infinitely small perturbations. Finally, our buffering concept provides a stringent definition of "buffering" on the level of systems and control theory, resulting in four absolute ratio scales for control performance that are suited to measure disturbance rejection and setpoint tracking, and both their static and dynamic aspects. CONCLUSION: Our concept of buffering provides a powerful mathematical tool for the quantitation of buffering action in all its appearances.

Animals↗

Using nitrogen-15 to quantify vegetative buffer effectiveness for sequestering nitrogen in runoff.

Previous studies have observed higher levels of soluble nutrients leaving vegetative buffers than entering them, suggesting that the buffers themselves are acting as a source rather than a sink by releasing previously stored nutrients. This study used 98 atom % (15)N-labeled KNO(3) at a rate of 5 kg ha(-1) to quantify buffer efficiency for sequestering new inputs of NO(-)(3)-N in an extensively grazed irrigated pasture system. Buffer treatments consisted of an 8-m buffer, a 16-m buffer, and a nonbuffered control. Regardless of the form of runoff N (NO(-)(3), NH(+)(4), or dissolved organic nitrogen [DON]), more (15)N was lost from the nonbuffered treatments than from the buffered treatments. The majority of the N attenuation was by vegetative uptake. Over the course of the study, the 8-m buffer decreased NO(-)(3)-(15)N load by 28% and the 16-m buffer decreased load by 42%. For NH(+)(4)-(15)N, the decrease was 34 and 48%, and for DON-(15)N, the decrease was 21 and 9%. Although the buffers were effective overall, the majority of the buffer impact occurred in the first four weeks after (15)N application, with the buffered plots attenuating nearly twice as much (15)N as the nonbuffered plots. For the remainder of the study, buffer effect was not as marked; there was a steady release of (15)N, particularly NO(-)(3)- and DON-(15)N, from the buffers into the runoff. This suggests that for buffers to be sustainable for N sequestration there is a need to manage buffer vegetation to maximize N demand and retention.

Adsorption↗

Sodium bicarbonate or multielement buffer via diet or rumen: effects on performance and acid-base status of lactating cows.

Our objective was to compare the influence of dietary NaHCO3 and a multielement buffer on ruminal acid-base status and lactation performance of dairy cows. Five ruminally fistulated, primiparous and multiparous lactating Holstein cows averaging 123 +/- 21 d postpartum were assigned randomly to treatments in a 5 x 5 Latin square with 3-wk experimental periods. Treatments were a basal diet without supplemental buffers, with 1.5% NaHCO3 or 1.5% multielement buffer, or with NaHCO3 or multielement buffer solutions poured into the rumen via cannula at 2 h postfeeding. Addition of either buffer to the diet reduced ruminal fluid hydrogen ion concentration from 0 to 6 h postfeeding; only NaHCO3 reduced ruminal fluid acidity when dosed via the cannula. Addition of buffers via ruminal cannula appeared to retard the reduction in ruminal fluid acidity that normally occurs from 6 to 12 h postfeeding; this may have been related to a feedback mechanism inhibiting salivary buffer secretion. Buffering capacity of ruminal fluid tended to increase with buffer addition; the increase was greatest during infusion of NaHCO3. The ruminal fluid buffer value index increased by 4 units for control cows from early (0 to 6 h) to late (6 to 12) postfeeding; smaller increases were noted for addition of multielement buffer. This index was not different for NaHCO3 during these two intervals. Milk yield and DMI were not affected by buffer addition. Although milk fat content tended to be higher with the multielement buffer than with NaHCO3, it was not accompanied by the expected alterations in ruminal acid-base status. Therefore, this increase may be related to systemic effects of specific minerals in the multielement buffer rather than to a more stable ruminal environment. Based on the ruminal fluid buffer value index, NaHCO3 tended to maintain the most stable ruminal acid-base status.

Acetates↗