Tetrodotoxin-sensitive uptake of ions and water by slices of rat brain in vitro

1. Exchange of (3)H(2)O with H(2)O takes place rapidly in incubated rat brain slices but at a lower rate in slices from infant brain than from adult brain. The temperature coefficient (Q(10)) of the exchange process, between 37 and 4 degrees C, is 1.76 with infant brain and 1.26 with adult brain. The exchange process is unaffected by the presence of ouabain or 2,4-dinitrophenol. 2. An approximately linear relationship exists between water uptake and the concentration of ATP in the incubated slices in the presence of various concentrations of glucose. Little or no change occurs in water uptake and ATP concentration in the presence of a glucose concentration exceeding 3mm. A linear relationship also exists between the water uptake and ATP concentration in the presence of 10mm-glucose and various concentrations of sodium l-glutamate but the line is parallel to that found with changed glucose concentrations and shifted in the direction of increased water uptake. A similar parallel relationship exists between water uptake and ATP concentration in the presence of 2,4-dinitrophenol, but the amount of water uptake is significantly smaller in the presence of 2,4-dinitrophenol than in its absence. 3. Copper chloride (0.3mm) or mercuric chloride (0.3mm) both increase water uptake and diminish the ATP concentration in slices. Sodium malonate (2mm) or sodium d-glutamate (10mm) has similar effects. 4. Substances, or conditions, affecting water uptake in incubated brain slices may be divided roughly into two classes in accordance with their effects on adenosine triphosphatase and membrane permeability, but there may be considerable lack of specificity.     

A simple model of the hydrophobic effect for molecular simulation of interfacial phenomena

A coarse-grained model for simulation of interfacial phenomena in aqueous systems has been developed. The model captures the hydrophobic effect by only considering the structure and cohesiveness of water. Monte Carlo (MC) simulations of water-oil mixtures show that low concentrations of oil are solvated with little perturbation of the hydrogen bonding network structure of the water, while high concentrations of oil are excluded altogether. Analysis of the water structure in the simulations indicates that the water molecules maintain close to four coordination in the presence of solutes and the distribution of bond angles is not markedly affected by the presence of solutes. MC simulations of an alkane oligomer in water and a poly(ethylene oxide) (PEO) oligomer in water indicate that the chains are quite flexible and also do not perturb the network structure of the water phase.     

Significance of enhanced morphological detection of Cryptosporidium sp. oocysts in water concentrates determined by using 4',6'-diamidino-2-phenylindole and immunofluorescence microscopy

Of 2,361 water concentrates analyzed for the presence of Cryptosporidium spp. oocysts between January 1992 and May 1998, 269 (11.4%) were positive, of which 235 (87.4%) were raw and 34 were final water concentrates. Of 740 oocysts enumerated in positive samples, 656 oocysts (88.7%) were detected in raw and 84 oocysts (11.3%) were detected in final water concentrates by using a commercially available fluorescein isothiocyanate-labeled anti-Cryptosporidium sp. monoclonal antibody and the nuclear fluorogen 4',6'-diamidino-2-phenylindole (DAPI). Of raw water positive samples, 66.8% had oocysts that contained nuclei, while 58.8% of final water samples had oocysts that contained nuclei. The most frequently identified oocysts had either no DAPI-positive nuclei and no internal morphology according to Nomarski differential interference-contrast microscopy (DIC) or four DAPI-positive nuclei together with internal contents according to DIC (39.5 and 32.8% of raw and 42.9 and 30.9% of final water positives, respectively). By use of the presence of DAPI-stained nuclei to support oocyst identification based upon oocyst wall fluorescence, 56.5% of oocysts were identified when at least one nucleus was present, while increasing the number of nuclei necessary for identification to four reduced the percentage identifiable to 32.8% in raw water concentrates. In final water concentrates, 51% of oocysts were identified using oocyst wall fluorescence and the presence of at least one nucleus, while increasing the number of nuclei necessary for identification to four reduced the percentage identifiable to 30.9%. By consolidating our identification criteria from the presence of at least one nucleus to the presence of four nuclei, we excluded approximately 20% of oocysts in either water type. Approximately 40% of oocysts detected in these United Kingdom samples were empty and could not be detected by alternative methods, including the PCR and fluorescence in situ hybridization.     

Significance of Enhanced Morphological Detection of Cryptosporidium sp. Oocysts in Water Concentrates Determined by Using 4′,6′-Diamidino-2-Phenylindole and Immunofluorescence Microscopy

Of 2,361 water concentrates analyzed for the presence of Cryptosporidium spp. oocysts between January 1992 and May 1998, 269 (11.4%) were positive, of which 235 (87.4%) were raw and 34 were final water concentrates. Of 740 oocysts enumerated in positive samples, 656 oocysts (88.7%) were detected in raw and 84 oocysts (11.3%) were detected in final water concentrates by using a commercially available fluorescein isothiocyanate-labeled anti-Cryptosporidium sp. monoclonal antibody and the nuclear fluorogen 4′,6′-diamidino-2-phenylindole (DAPI). Of raw water positive samples, 66.8% had oocysts that contained nuclei, while 58.8% of final water samples had oocysts that contained nuclei. The most frequently identified oocysts had either no DAPI-positive nuclei and no internal morphology according to Nomarski differential interference-contrast microscopy (DIC) or four DAPI-positive nuclei together with internal contents according to DIC (39.5 and 32.8% of raw and 42.9 and 30.9% of final water positives, respectively). By use of the presence of DAPI-stained nuclei to support oocyst identification based upon oocyst wall fluorescence, 56.5% of oocysts were identified when at least one nucleus was present, while increasing the number of nuclei necessary for identification to four reduced the percentage identifiable to 32.8% in raw water concentrates. In final water concentrates, 51% of oocysts were identified using oocyst wall fluorescence and the presence of at least one nucleus, while increasing the number of nuclei necessary for identification to four reduced the percentage identifiable to 30.9%. By consolidating our identification criteria from the presence of at least one nucleus to the presence of four nuclei, we excluded approximately 20% of oocysts in either water type. Approximately 40% of oocysts detected in these United Kingdom samples were empty and could not be detected by alternative methods, including the PCR and fluorescence in situ hybridization.     

Anticipatory behaviour and stereotypical behaviour in farmed mink (Mustela vison) in the presence,absence and after the removal of swimming water

Captive animals are often provided with cage enrichments in order to improve their welfare. Swimming water is an often-discussed requirement for farmed mink. The present study aimed to give insight into the value of swimming water for farmed mink by measuring anticipatory and stereotypical behaviour in subjects raised and housed in the presence and absence of swimming water and in subjects that were deprived of swimming water.The major findings of the present experiments are that: (1) there was no significant difference in anticipatory behaviour between subjects reared and housed in the presence or absence of swimming water; nor was there a significant difference in anticipatory behaviour after removing the water for 2.5 weeks, (2) there was no significant difference in stereotypical behaviour in winter in the presence, absence and after 2.5 months deprivation of swimming water, and finally, (3) there was no significant correlation between anticipatory activity and stereotypical behaviour.The results suggest that mink might not experience consequences, in the sense of increased reward-sensitivity or stereotypy levels, due to the absence of swimming water if they never experienced this incentive before, and that swimming water and an empty bath, such as used in this experiment, might be equally valued incentives for mink.     

Bioreduction of a carbon–nitrogen double bond using immobilized baker's yeast’a first report

Immobilized baker's yeast entrapped in calcium alginate beads efficiently reduces N-benzylidinemethylamine to N-methylbenzylamine in hexane at 37°C and tetrahydrofuran (THF) at 30°C in the presence of 18-crown-6, while in the presence of water as cosolvent and glucose as an additive N-benzylidinemethylamine undergoes decomposition. Benzaldoxime in a hexane–water (1:9) solvent system containing glucose as an additive is reduced to N-benzylhydroxylamine. On using an ethanol–water (1:1) solvent system, benzaldoxime is converted to benzyl alcohol and in hexane, benzene, THF, hexane–water (1:1) or acetonitrile–water (1:1) solvent systems, or using dried baker's yeast in different solvent systems, transformation of benzaldoxime does not occur.     

Bioreduction of a carbon–nitrogen double bond using immobilized baker''s yeast’a first report

Immobilized baker's yeast entrapped in calcium alginate beads efficiently reduces N-benzylidinemethylamine to N-methylbenzylamine in hexane at 37°C and tetrahydrofuran (THF) at 30°C in the presence of 18-crown-6, while in the presence of water as cosolvent and glucose as an additive N-benzylidinemethylamine undergoes decomposition. Benzaldoxime in a hexane–water (1:9) solvent system containing glucose as an additive is reduced to N-benzylhydroxylamine. On using an ethanol–water (1:1) solvent system, benzaldoxime is converted to benzyl alcohol and in hexane, benzene, THF, hexane–water (1:1) or acetonitrile–water (1:1) solvent systems, or using dried baker's yeast in different solvent systems, transformation of benzaldoxime does not occur.     

Evaluation of a water concentration method for the detection of infectious pancreatic necrosis virus in a fish hatchery

A virus-adsorption-elution (viradel) method using electropositive microporous filters (Virosorb 1-MDS) was used for the concentration and détéction of infectious pancreatic necrosis virus (IPNV) at a hatchery with a past history of IPNV infection. Samples of fish tissue and unconcentrated water were also examined for the presence of IPNV. Virus was isolated from three of the nine concentrated water samples taken from various locations at the hatchery. Virus was also isolated from pooled fish tissues, but not from unconcentrated water samples. Representative numbers of viruses isolated from water and fish tissues were examined for virus neutralization in the presence of anti-IPNV amiserum; resistance to acid, ether, and heat; and replication in the presence of 5-bromo-2'-deoxyuridine (BUDR). When tested by dot-immunobinding assay using a panel of monoclonal antibodies (MABs), the viral isolates were found to belong to the West Buxton serotype of IPNV.     

FLAVOR THRESHOLD AS AFFECTED BY INTERACTION AMONG THREE DAIRY‐RELATED FLAVOR COMPOUNDS*

The hypothesis of the present study was that the presence of one or more flavor compounds in a matrix would alter the perception (either enhance or suppress) of individual compounds as compared to a single flavor compound in a matrix. Thresholds were established for the flavor compounds (diacetyl, hexanal and δ‐decalactone) within each matrix; for each compound in the presence of a second compound; and for each compound in the presence of the remaining two compounds. An ascending order 2‐alternative forced choice test with 12 panelists was used for the evaluations. Results showed that diacetyl in water was suppressed by the presence of hexanal, but the presence of δ‐decalactone with hexanal reversed the trend. In the binary skim milk solutions diacetyl perception was enhanced, while being suppressed in the ternary solution. The perception of hexanal was enhanced in water (binary and ternary solutions). Hexanal and δ‐decalactone were suppressed in skim milk (binary and ternary solutions). δ‐Decalactone showed similar trends as hexanal in water, except that it was suppressed in the presence of diacetyl.     

A study of water binding in lyophilized viral vaccine systems

Differential scanning calorimetry (DSC) has been used to estimate the amount of bound water in freeze-dried live measles vaccine in the presence of two cryoprotective mixtures, with and without sucrose. The purpose of the work was to see if a relationship exists between the amount of bound water in viral systems, the nature of the freeze-drying process, and the final stability of the virus preparations.It was shown that although the presence of sucrose in protective agents improves the retention of activity, the limiting amount of bound water (that for which the heat of fusion is zero) in the rehydrated samples was not related to the presence of sucrose, but depended on the method of freeze-drying.     

Absence of association between total heterotrophic and total coliform bacteria from a public water supply

Heterotrophic plate counts (HPC) and total coliforms (TC) are two major microbial indicators that are used to monitor the potability of water. Although the presence of heterotrophs has been hypothesized to predict the presence of TC, there have been few documented reports. Intensive sampling of raw, treated effluent and distribution water from a public water supply serving 400,000 people provided an opportunity to study the relationship between these two indicator groups of bacteria. A total of 26,158 samples were analyzed, including 12,970 from 1986 and 13,188 from 1985. There were 13,429 samples from the distribution system, 5,524 from treatment effluents, and 7,205 from raw water. The associations between HPC and TC were made on both a hits-and-misses and numerical comparison (CFU per milliliter) basis. The periodicity of the two indicators was also analyzed to determine whether the presence of one group could predict the presence of the other. Atypical bacteria were also related to the presence of these two indicator bacteria. Venn diagrams and nonparametric statistics revealed the following correlation coefficients for HPC and TC for 1985 and 1986 combined: raw water r = 0.45, treated effluent r = 0.06, and distribution system r = 0.10. Atypical bacteria showed a similar relationship with HPC. There was no predictive periodicity for HPC and TC within +/- 10 days of isolation of each other. Therefore, in a 2-year survey of a public water supply, the presence of HPC had a low correlation coefficient with TC, as determined by hits-and-misses and numerical comparisons. The enumeration of one group was found to be independent of the other.     

Absence of association between total heterotrophic and total coliform bacteria from a public water supply.

Heterotrophic plate counts (HPC) and total coliforms (TC) are two major microbial indicators that are used to monitor the potability of water. Although the presence of heterotrophs has been hypothesized to predict the presence of TC, there have been few documented reports. Intensive sampling of raw, treated effluent and distribution water from a public water supply serving 400,000 people provided an opportunity to study the relationship between these two indicator groups of bacteria. A total of 26,158 samples were analyzed, including 12,970 from 1986 and 13,188 from 1985. There were 13,429 samples from the distribution system, 5,524 from treatment effluents, and 7,205 from raw water. The associations between HPC and TC were made on both a hits-and-misses and numerical comparison (CFU per milliliter) basis. The periodicity of the two indicators was also analyzed to determine whether the presence of one group could predict the presence of the other. Atypical bacteria were also related to the presence of these two indicator bacteria. Venn diagrams and nonparametric statistics revealed the following correlation coefficients for HPC and TC for 1985 and 1986 combined: raw water r = 0.45, treated effluent r = 0.06, and distribution system r = 0.10. Atypical bacteria showed a similar relationship with HPC. There was no predictive periodicity for HPC and TC within +/- 10 days of isolation of each other. Therefore, in a 2-year survey of a public water supply, the presence of HPC had a low correlation coefficient with TC, as determined by hits-and-misses and numerical comparisons. The enumeration of one group was found to be independent of the other.     

Supercooling characteristics of isolated peach flower bud primordia

The amount of unfrozen water in dormant peach (Prunus persica [L.] Batsch, cv Redhaven) flower buds, isolated primordia, and bud axes was determined during freezing using pulse nuclear magnetic resonance methods. Differential thermal analysis studies were conducted on whole buds and isolated primordia in the presence of ice nucleation. The results showed that some of the water in isolated primordia remained supercooled in the presence of ice nucleation. Although most tissue water froze (57.5%) following ice nucleation at −2.5°C, a considerable amount of water was found to supercool. In the presence of ice nucleation, increased hydration of isolated primordia resulted in the elimination of the supercooling characteristic. The structural integrity of isolated primordia appeared to be essential for supercooling.     

KCl potentiation of the virucidal effectiveness of free chlorine at pH 9.0.

In studies at 5 degrees C and pH 9.0, poliovirus 1 was inactivated about 15 times more rapidly by free chlorine (FC) in purified water in the presence of 1,262 mg of KCl per liter (approximately 0.0169 M) than in the absence of KCl. In the presence of 526 mg of KCl per liter, the virus was inactivated about seven times more rapidly by FC than in the absence of KCl. At a level of 21 mg/liter, KCl did not significantly potentiate the virucidal activity of FC in purified water. Although poliovirus 1 was inactivated almost three times more rapidly by FC in borate-buffered purified water than in purified water, the presence of the buffer did not alter the extent of potentiation by KCl. Most of FC exists as OCl- at pH 9.0. Tap water has been shown to markedly potentiate the polivirucidal effectiveness of FC at pH 9.0. For the same degree of virucidal potentiation of FC at this pH, a considerably greater quantity of KCl was required in purified water than the total salt content that appeared to be present in the tap water.     

A smoke-derived butenolide alleviates HgCl2 and ZnCl2 inhibition of water uptake during germination and subsequent growth of tomato--possible involvement of aquaporins

Aquaporins, concentrated in zones of cell division and enlargement, play a major role in governing the movement of water between neighboring cells during seed germination. The enhanced germination and growth found with smoke-water and butenolide could be the result of better water uptake, suggesting the involvement of aquaporins. The effects of butenolide, known aquaporin inhibitors (HgCl(2) and ZnCl(2)), along with several chemical agents known to reverse the inhibitory effects of mercuric chloride on the activity of aquaporins were tested. Seedlings raised in the presence of butenolide had higher moisture content (93%) compared to those imbibed in water only (85%). This suggests enhanced activity of aquaporins. The presence of aquaporin inhibitors (HgCl(2) and ZnCl(2)) reduced seedling water content and altered root development. The presence of HgCl(2) (10, 20 or 30muM) reduced the percentage imbibition of seeds by 11-12%. A corresponding gradual decline, from 17.5% (10muM) to 22.6% (30muM) (p0.05), in the root length was recorded. Addition of dithiothreitol (DTT, 500muM), beta-mercaptoethanol (ME, 250muM) and butenolide (0.1muM) along with the HgCl(2) overcame the observed inhibitory effects. The presence of ZnCl(2) (12.5 or 25muM) affected percentage imbibition as well as root length, which was reversed to some extent following addition of the butenolide. Though zinc chloride-mediated inhibition remained unaffected by the presence of DTT and ME, the butenolide reversed the effect. These results are interesting as they suggest additional avenues of research for uncovering the profound effect butenolide has on germination and growth.     

Water channels in platelet volume regulation

The regulation of platelet volume significantly affects its function. Because water is the major molecule in cells and its active transport via water channels called aquaporins (AQPs) have been implicated in cellular and organelle volume regulation, the presence of water channels in platelets and their potential role in platelet volume regulation was investigated. G-protein-mediated AQP regulation in secretory vesicle swelling has previously been reported in neurons and in pancreatic acinar cells. Mercuric chloride has been demonstrated to inhibit most AQPs except AQP6, which is stimulated by the compound. Exposure of platelets to HgCl(2)-induced swelling in a dose-dependent manner, suggesting the presence of AQP6 in platelets. Immunoblot analysis of platelet protein confirmed the presence of AQP6, and also of G(αo), G(αi-1) and G(αi-3) proteins. Results from this study demonstrate for the first time that in platelets AQP6 is involved in cell volume regulation via a G-protein-mediated pathway.     

Effects of NaCl and Na2SO4 on Water Relations ofPlantago maritima (L.) andPlantago lanceolata (L.)

Transpiration and water absorption rates, stomatal and cuticular resistances to water vapour diffusion, were measured onPlantago maritima (halophyte) andPlantago lanceolata (glycophyte) grown in the presence of NaCl or Na₂SO₄. Water absorption was reduced in the presence of Na₂SO₄ and transpiration rate was increased when NaCl was added to the nutrient solution. The glycophyte daily water balance was more disturbed than that of the halophyte in the presence of Na₂SO₄.     

The combined effect of environmental factors and neighbouring populations on the distribution and abundance of Arvicola terrestris. An approach using rule-based models

We investigated how the interplay between environmental factors and presence of neighbouring populations determines the distribution and abundance of a small, endangered rodent, the water vole ( Arvicola terrestris ). We studied thriving and non-fragmented populations of water voles in the absence of their main predator, the introduced American mink ( Mustela vison ). A low degree of population fragmentation, such as the one characterizing the studied populations, was probably typical of water voles before their decline started. We found that under these conditions water voles' distribution is mainly determined by three environmental factors: presence of freshwater, adequate food, and cover. Variance in other factors is well tolerated by water voles. We obtained this result by the use of rule-based models in two separate areas. The two models correctly classified 81% and 83% of the observed cases, respectively. When optimised on one area and cross-validated on the other area the performance of the models was still high (73% and 79%) indicating that the models were robust and generalizable. We also found that the density of animals was lower in sub-optimal than in optimal habitat. We then tested the hypothesis that the number of neighbouring colonies determines the probability of finding voles in a given section. We found that the presence of nearby colonies was an important factor in determining the presence of water voles in sub-optimal habitat, while isolated patches of suitable habitat were less likely to host water voles. These observations suggest the possible presence of a source-sink dynamic, where an optimal habitat acts as a source for populating sub-optimal habitats that may be considered a sink habitat. These findings are discussed in the context of water vole conservation.     

Fishing spiders, green sunfish, and a stream-dwelling water strider: male-female conflict and prey responses to single versus multiple predator environments

Many studies have experimentally addressed the effects of a particular predator species on prey behavior. In nature, however, prey frequently face multiple species of predators that often vary in their predatory mode and in their level of predation risk. Relatively few studies have considered prey responses under these complex conditions. In Kentucky, the stream-dwelling water strider (Aquariusremigis) coexists with many potentially dangerous predators, two of which are the green sunfish (Lepomiscyanellus) and the fishing spider (Dolomedesvittatus). Green sunfish occupy stream pools and attack water striders from below. In contrast, fishing spiders hunt along stream shorelines where they perch on overhanging vegetation or rocks and attack water striders near shore. We compared how A. remigis individuals respond to these two very different predators in pools with one or both predators. The presence of sunfish in pools had strong effects on male water strider behavior, including increased use of three types of refuge from sunfish (riffles, climbing out of the water, sitting on the water but at the edges of pools), decreased activity and a decreased number of aggressive males on the water. Spiders also influenced water strider behavior; male water striders avoided spiders by shifting away from the edges of pools. Comparisons of the effects of the two predator species showed that in general, antipredator responses by male water striders were stronger in pools with fish alone than in those with spiders alone. In the presence of both predators, male water strider behavior (microhabitat use and activity) was generally similar to behavior in the presence of fish alone. In contrast, female water striders showed no significant response to the presence of sunfish, and little response to the presence of spiders. This lack of response could be because females spent much of their time in refuges even in the absence of predators (apparently hiding from harassment by males). Both spiders and fish caused decreases in water strider mating activity. The presence of fish reduced both the number of matings per pool (mating frequency), and mean mating durations. Spiders induced a decrease in mean mating duration, but not in mating frequency. The largest reductions in mating activity occurred in pools with both predators present. Pools with either spiders or fish alone suffered 15–20% water strider mortality during our experiment (versus no mortality in predator-free pools). Extant theory suggests that when prey face conflicting microhabitat responses to two predators (as in this study), the predators should have facilitative effects on predation rates (i.e., prey that avoid one predator are often killed by the other and vice versa). Mortality rates in pools with both predators present, however, were not significantly different from that predicted by a null model of multiple predator effects. The lack of predator facilitation can be explained by the compensatory reductions in water strider activity and mating activity in the presence of both predators. Received: 26 August 1996 / Accepted: 12 June 1998     

Macromolecular source as dependent on osmotic pressure and water source: effects on bovine in vitro embryo development and quality

This study evaluated the protective effect of protein, as dependent on osmolarity, and the quality of water sources used to prepare embryo culture media. In Experiment 1, two concentrations of NaCl were used to obtain culture media with normal (280 mOSM) and low (245 mOSM) osmolarity, each supplemented with either bovine serum albumin (BSA) or polyvinyl alcohol (PVA). Low osmolarity improved blastocyst rates in the presence of BSA (P < 0.01) and tended to do it in medium containing PVA (P < 0.07). Furthermore, low osmolarity allowed PVA to increase inner cell mass (ICM) numbers and ICM/total cell rate (P < 0.05), while trophectoderm (TE) and total cell counts tended to decrease (P < 0.08). In Experiment 2, culture media were prepared with two water sources (Milli-Q and Sigma-W3500-) in combination with BSA or PVA. Both water sources yielded similar embryo development rates, but in the presence of BSA, Milli-Q water produced embryos with increased ICM/total cells rates (P < 0.05). On the contrary, Sigma water tended to increase trophectoderm cell counts (P < 0.08). In conclusion, the present study showed that low osmolarity is beneficial to embryo development and combinations of macromolecule and osmolarity influence trophectoderm differentiation. Both Milli-Q and Sigma supported embryo development at comparable rates, although in the presence of BSA, blastocysts obtained in the medium prepared with Milli-Q water had superior quality in terms of ICM/total cells rates.