Increased oxygen consumption during the uptake of water by the eversible vesicles of Petrobius brevistylis

The oxygen consumption of individuals of Petrobius brevistylis has been measured before, during, and after the uptake of fluid by the insects eversible abdominal vesicles. There was no difference in consumption between fresh animals and those that had been dehydrated or those that had been allowed to rehydrate after an exposure to a NaCl solution. However, during the process of taking up fluid there was over a 200% increase in the respiration rate. While the increase in the rate of oxygen consumption was related to the concentration of the fluid from which the animals were taking up water, the increase in the amount of oxygen consumed was related to the concentration of the substrate solution and the amount of uptake. Part of the oxygen increase is ascribed to the posture adopted by the animals during the uptake of fluid and the remainder to the active transport processes that may be occurring.     

NaCl intake and preference threshold of spontaneously hypertensive rats.

Both male and female spontaneously hypertensive (SH) rats have an appetite for NaCl solution. The appetite is present when a choice is offered between distilled water and either isotonic or hypertonic (0.25 M) NaCl solution to drink. Total fluid intake (water plus NaCl solution) was greater for SH rats than for controls while food intakes (g/100 g body wt/day) of SH rats were not different from controls. Mean body weight of SH rats was always less than that of controls. The appetite for NaCl solution was accompanied by a significant reduction in preference (detection) threshold. SH rats could detect the difference between distilled water and NaCl solution when the concentration of the latter was 12 mEq/liter compared to a control threshold of 30 mEq/liter. The NaCl appetite and reduced NaCl preference threshold induced by spontaneous hypertension is in marked contrast to the NaCl aversion induced by other types of experimentally induced hypertension in rats. The mechanism or mechanisms responsible for these differences remain for further study.     

A New Method of Silver Impregnation for Nerve Cells and Fibers

Fragments of tissue, immediately after death, are fixed in Debaisieux's modification of the Duboscq-Brazil picro-aceticformol fluid, and treated as follows: Hydrate by soaking 2-6 hr. in distilled water with 30 drops of cone. NH₄OH per 100 cc. Freeze and cut sections about 25μ in thickness. Bleach sections about 15 min. in ammoniacal water (52 drops cone. NH₄OH per 100 cc. water). Transfer to 20% AgNO₃ solution and heat at 45° C. till light brown. Add cone. NH₄OH drop by drop till the Ag precipitates and then redisolves into an opalescent solution. Pour solution and sections into a little distilled water and transfer sections quickly to formaldehyde solution (3 cc. formalin to 100 cc. water). Dip sections in distilled water and transfer to 1% aqueous gold chloride till deep blue. Place for about 10 minutes in 5% aqueous sodium thiosulfate solution for fixing and clearing. Wash thoroly in tap water, dehydrate and mount. Special directions are given for applying this technic to delicate material such as insects, and for use with serial sections.     

A C-terminal aldehyde analog of the insect kinins inhibits diuresis in the housefly

The insect kinins are present in a wide variety of insects and function as potent diuretic peptides in flies. A C-terminal aldehyde insect kinin analog, Fmoc-RFFPWG-H (R-LK-CHO), demonstrates stimulation of Malpighian tubule fluid secretion in crickets, but shows inhibition of both in vitro and in vivo diuresis in the housefly. R-LK-CHO reduced the total amount of urine voided over 3 h from flies injected with 1 microL of distilled water by almost 50%. The analog not only inhibits stimulation of housefly fluid secretion by the native kinin Musdo-K, but also by thapsigargin, a SERCA inhibitor, and by ionomycin, a calcium ionophore. The activity of R-LK-CHO is selective, however, as related C-terminal aldehyde analogs do not demonstrate an inhibitory response on housefly fluid secretion. The selective inhibitory activity of R-LK-CHO on housefly tubules represents an important lead in the development of environmentally friendly insect management agents based on the insect kinins.     

Factors affecting the lytic susceptibility of some marine and terrestrial bacteria

Eighteen gram-negative marine bacteria and two terrestrial species, Escherichia coli and Pseudomonas aeruginosa, were examined for their sensitivity to lysis in distilled water after exposure to a salt solution containing a sea water concentration of Mg2+ (0.05 M) or to 0.5 M NaCl. A spectrum of lytic susceptibility was observed among the marine bacteria ranging from those organisms which lysed in distilled water after exposure to the Mg2+-containing solution, through organisms which could be sensitized to lysis by washing with the NaCl solution, to organisms which failed to lyse in distilled water even after having been washed with a solution of 0.5 M NaCl. Pseudomonas aeruginosa and E. coli fell within this spectrum, the former being capable of being induced to lyse in distilled water by washing with 0.5 M NaCl, while the latter failed to lyse in distilled water after this treatment. It was thus concluded that no overall distinction could be made between marine and terrestrial bacteria on the basis of the sensitivity of the two groups of organisms to lysis in freshwater. Quite large decreases in optical density and increases in the release of ultraviolet-absorbing material took place when cells preexposed to the Mg2+-containing solution or to 0.5 M NaCl were subsequently suspended in distilled water even though in some cases no loss of cell numbers could be detected. In most cases two to three times as much K+ as Na+ and 1/10 to 1/100 as much Mg2+ was required to prevent these changes. For three of the marine bacteria and P. aeruginosa grown in a terrestrial type medium little difference in the requirements for Na+ and K+ to prevent the optical density changes was noted. For P. aeruginosa grown in a marine type medium, cells required more K+ than Na+ to prevent these changes.     

Fluid secretion by single isolated Malpighian tubules of the house cricket, Acheta domesticus, and their response to diuretic hormone

ABSTRACT. A simple modification of the Ramsay method is described for the isolation in vitro of Malpighian tubules from Acheta domesticus. Isolated tubules continue to secrete for many hours in a simple Ringer solution. The rate of secretion is linearly related to the length of tubule within the bathing fluid.
Secretion rates during the immediate post-equilibration period relate to the physiological state of the donor insect. Highest rates are recorded in tubules removed from animals during the scotophase when the crickets are most actively feeding. Starvation reduces the rate of secretion and when previously starved insects are given access to food the rate increases. These results are consistent with variations in the haemolymph titre of DH that are induced by feeding.
Tubule fluid secretion is stimulated by either CC extract (1 gland pair/50 μ1) or dibutyryl cAMP (10^-3M). CC extracts increase the rate of fluid secretion by c. 200 pl/mm/min for up to 6 h after the tubules have been isolated. The response varies little with the age or physiological state of the donor insect. However, tubules from newly moulted insects secrete at very low rates and respond poorly to CC extract, although they are stimulated by cAMP. The significance of these results is discussed with particular reference to the use of Acheta tubules as a bioassay preparation for DH.     

Inhibition of thirst when dehydrated rats drink water or saline

The present experiments sought to identify the physiological signals that inhibit thirst when dehydrated rats drink water or NaCl solution. Rats were deprived of drinking fluid but not food overnight. When allowed to drink again, the dehydrated animals consumed water or saline (0.05 M, 0.10 M, 0.15 M, or 0.20 M NaCl solution) almost continuously for 5-8 min before stopping. The volumes consumed were similar regardless of which fluid they ingested, but blood analyses indicated that increased plasma osmolality and decreased plasma volume, or both, still remained when drinking terminated. These results suggest that the composition of the ingested fluid is less significant than its volume in providing an early signal that inhibits thirst and fluid consumption by dehydrated rats. Analyses of the gastrointestinal tracts revealed that the cumulative volume in the stomach and small intestine correlated highly with the amount consumed regardless of which fluid was ingested. These and other results suggest that the volume of fluid ingested by dehydrated rats is sensed by stretch receptors detecting distension of the stomach and small intestine, which provide an early inhibitory stimulus of thirst.     

Motility of Mugil cephalus L. spermatozoa in coelomic fluid, seminal fluid and saline media

The aim of this study was to assess the motility duration of Mugil cephalus when exposed to seminal fluid, coelomic fluid and saline media. Hypo-osmotic activation medium (distilled water containing bovine serum albumin 10 or 30 mg ml⁻¹) did not trigger sperm motility. Saline solution containing 500 m m NaCl, 3.1 m m KCl, 0.2 m m Tris, 3.4 m m CaCl₂, pH 7.5 initiated the sperm activation and the motility lasted for more than 2 min. Coelomic fluid showed an inhibitory effect for triggering the motility of spermatozoa. Higher salinity increases the motility duration of sperm. The optimum motility duration was shown in salinity 32 psu.     

The protective effect of sodium chloride against thermal destruction of the phage-forming mechanism in staphylococci

Staphylococci which have been allowed to grow rapidly in a favorable environment and subsequently have been maintained in a resting state characteristically produce a sharp rise in [phage] (activity) titre when added to phage. This capacity is quickly lost when the cells are suspended in distilled water and are exposed to 44 degrees C. for a period of 15 minutes; at the same time the viable count drops to approximately 1 to 3.5 per cent of the initial value. 1 M NaCl protects "activated" cells against thermal destruction and preserves the phage-augmenting property. "Non-activated" staphylococci in distilled water suspension do not show this thermolability.     

Identification and characterization of functional aquaporin water channel protein from alimentary tract of whitefly, Bemisia tabaci

Some hemipteran xylem and phloem-feeding insects have evolved specialized alimentary structures or filter chambers that rapidly transport water for excretion or osmoregulation. In the whitefly, Bemisia tabaci, mass movement of water through opposing alimentary tract tissues within the filter chamber is likely facilitated by an aquaporin protein. B. tabaci aquaporin-1 (BtAQP1) possesses characteristic aquaporin topology and conserved pore-forming residues found in water-specific aquaporins. As predicted for an integral transmembrane protein, recombinant BtAQP1 expressed in cultured insect cells localized within the plasma membrane. BtAQP1 is primarily expressed in early instar nymphs and adults, where in adults it is localized in the filter chamber and hindgut. Xenopus oocytes expressing BtAQP1 were water permeable and mercury-sensitive, both characteristics of classical water-specific aquaporins. These data support the hypothesis that BtAQP1 is a water transport protein within the specialized filter chamber of the alimentary tract and functions to translocate water across tissues for maintenance of osmotic pressure and/or excretion of excess dietary fluid.     

Taste sensitivity in the embryo of the domestic fowl

Two experiments were carried out to investigate the sense of taste in embryos of the domestic fowl. In the first, four taste substances; NaCl, HCl, glucose and SOA were diluted with distilled water and the response was compared with that to distilled water alone. No significant effects of taste were found. In the second experiment five taste substances: HCl, fructose, NaCl, KCl and quinine were diluted with fluids normally imbibed by the embryo: amniotic and/or allantoic fluid taken from other eggs. These solutions and also distilled water were compared with egg-fluid alone. A highly significant effect of the five solutions was found showing that the taste system becomes functional before the time of hatching. Distilled water produced on an unexpectedly large response in the embryo; possible reasons for this are discussed.     

A Ribonuclease-Gallocyanin Stain for Sexing Skin Biopsies

Skin biopsies for sexing can be fixed best in 10-15% aqueous formalin or this solution saturated with HgCl₂. Bouin's fluid and all chromate mixtures should be avoided. Celloidin-paraffin double embedding is recommended but not essential. Sections are brought to water, mercurial residues removed if necessary, and then washed in distilled water. They are incubated at 37°C in a ribo-nuclease solution: approximately 1 mg of ribonuclease powder (Light's) in 100 ml of glass-distilled water; boiled 3-5 sec after dissolving, and kept in a refrigerator (usable about a week). The sections are rinsed and incubated at 37°C overnight in gallocyanin-chromalum (Einarson, 1951) made as follows: Dissolve 5 gm of chromalum in 100 ml of distilled water, add 0.15 gm of gallocyanin, shake thoroughly, heat slowly and boil 5 min; cool, filter, and wash through the filter with distilled water until the filtrate reaches 100 ml. This solution is usable at once and keeps at least a month. Sections should be dipped in acid alcohol to clean (optional), but no attempt made to differentiate them, and washed in tap water. Dehydration, clearing and covering complete the process. The method is nearly as precise as the Feulgen and more convenient and reliable for routine use on miscellaneous material.     

Differential Staining Of Tissue In The Block With Picric Acid And The Feulgen Reaction

The authors have found a modification of the Feulgen reaction to be a satisfactory stain for tissue in the block.

Pieces of fresh mammalian tissue not thicker than 5 mm. are fixed for approximately 48 hours at 25° C. in a mixture of equal parts of 5% aqueous sulfosalicylic acid and saturated aqueous picric acid. They are washed for 30 minutes in three ten-minute changes of distilled water and placed in Feulgen's staining solution diluted to one-half strength with distilled water. The staining solution is allowed to act for 24 hours (2 to 3 mm. thick blocks) up to 48 hours for 5 mm. thickness. After staining, the specimens are transferred to a mixture of sodium bisulfite, 0.5 g. and N hydrochloric acid, 5 ml. in' 100 ml. of distilled water. Two changes of IS to 30 min. each in the acid sulfite are given and these are followed by dehydration through 50%, 70% and 95% alcohol. One to two hours are allowed for each change except the last 95%, in which the stained tissue is allowed to remain overnight. The dehydration is completed in two changes of absolute alcohol with subsequent clearing in xylene and embedding in paraffin. Sections may be cut 10 μ or other thickness desired, mounted on slides, paraffin removed, and covered in the usual manner. Nuclei stain reddish violet against a lemon yellow background when the stain is typical. Orange G, 200 mg. per 100 ml. may be added to the fixing fluid if a more polychromatic effect is desired.     

Urinary kallikrein excretion after potassium adaptation in the rat

24-h urinary kallikrein excretion in male Sprague-Dawley rats was measured before and after 14 days with 100 mM potassium chloride as drinking fluid ad libitum. Urinary kallikrein excretion increased in K+-adaptation. The increase was greater when the rats were given distilled water rather than 100 mM sodium chloride to drink prior to the potassium chloride. The urinary potassium excretion increased in all rats studied. The urinary sodium excretion, urine volume and fluid intake increased significantly in rats that had distilled water to drink prior to the KCl. In marked contrast, when rats were offered NaCl prior to KCl, the urinary sodium excretion was unaffected while the urine volume and fluid intake decreased significantly. This study shows that prior NaCl intake abolishes the natriuretic and diuretic effects of KCl load and only suppresses the increase in urinary kallikrein excretion. This suggests that K+ secretory activity at the distal tubules is the major determinant of the release of renal kallikrein in the rat.     

Viscoelastic properties of human tracheobronchial mucin in aqueous solution

Human tracheobronchial mucin isolated from cystic fibrosis patients (CF HTBM) was purified using a combination of gel filtration and density gradient centrifugation. The resulting mucin was fractionated to reduce polydispersity and to facilitate studies of the molecular weight dependence of mucin viscoelasticity in concentrated solution. The viscoelastic properties of CF HTBM were examined in distilled water, 0.1M salt solutions and chaotropic solvents. In controlled strain experiments (strain ≥ 5%) with increasing mucin concentration, a crossover from sol to gel behavior is observed. The gel strength, as measured by the magnitude of the storage modulus at comparable mucin concentrations, is greatest for distilled water, intermediate for 0.1M NaCl, and lowest far 6M GdnHCl. In distilled water, high molecular weight mucin undergoes a sol-gel transition at ～ 12 mg/mL, and shows evidence of a plateau modulus at higher concentrations. The storage and loss moduli of concentrated high molecular weight fractions in 6M GdnHCl exhibit a power law dependence on frequency typical of weak gels near the sol–gel transition at 20 mg/mL. Similar rheology is observed in 0.1M NaCl and 0.091M NaCl/3 mM CaCl₂, but with evidence for additional weak associations at low frequency. The power law exponent in these systems is 0.70 ± 0.02, in good agreement with prediction for networks formed by a percolation mechanism. Low molecular weight fractions in these solvents exhibit a fluid-like viscoelastic response. However, low molecular weight mucin in distilled water shows a strain-dependent increase in elasticity at low frequency indicative of weak intermolecular associations. Comparison of the rheological behavior of CF HTBM with our earlier studies of ovine submaxillary mucin lends support to the idea that carbohydrate side-chain interactions are important in the gelation mechanism of mucins. © 1995 John Wiley & Sons, Inc.     

Role of abscisic acid in the closure of stomata in some arid zone plant species

Initially closed stomata in isolated leaf epidermal peelings ofCalotropis procera, Prosopis cineraria andTephrosia purpurea, opened when incubated in distilled water in light or in darkness. Water-induced open stomata closed down on incubation in abscisic acid (0.01 ppm) solution within half an hour. Sodium chloride induced the opening of stomatal aperture. NaCl-induced open stomata also completely closed down when incubated in abscisic acid solution, but open again when transferred to NaCl.     

The effects of temperature and housing on water balance in a burrowing mouse,Peromyscus polionotus

Summary The effects of temperature and humidity on the water balance of two races ofPeromyscus polionotus were compared to individuals of the same race given moist sand to burrow in. Temperature had the greatest effect on fluid consumption. A change from 20 to 30°C resulted in a significant decrease in the amount of water and 0.4 M NaCl solution consumed. Generally, there was no difference in the amount of water or 0.4 M NaCl consumed at 20°C under different housing conditions, but at 30°C both races consumed significantly more fluids at the low humidity than when occupying a burrow. Considering all types of housing, temperature had a significant effect on a mouse's ability to maintain body weight when stressed with salt water. Irrespective of temperature, weight loss was less and survival greater while drinking salt water for mice in a burrow (92% survival) compared with the high (77% survival) and low (67% survival) humidity conditions.     

NaCl溶液对贵阳腐霉菌丝生长、孢子囊产生及灭蚊能力的影响

研究不同浓度NaCl溶液对贵阳腐霉菌丝生长、产孢能力及灭蚊能力的影响,探讨贵阳腐霉对NaCl溶液的耐受性,为其在含盐水体中控制蚊虫种群的应用提供理论依据。结果表明：低浓度NaCl溶液可以促进贵阳腐霉菌丝生长及孢子囊形成,高浓度NaCl溶液则抑制菌丝生长及孢子囊形成。于0.01%浓度NaCl溶液中培养24h菌丝生长速度最快,产孢子囊数量最多,与对照相比差异显著。随着NaCl浓度升高,菌丝及孢子囊形成均受影响。菌丝生长速度降低,孢子囊形成数量减少。在0.75% NaCl溶液中贵阳腐霉的菌丝生长严重受抑制,且产孢子囊能力为零。灭蚊实验表明,0.01% NaCl溶液中贵阳腐霉的灭蚊能力在72–96h时均比蒸馏水对照组强,但二者未有显著差异,120h蒸馏水对照组灭蚊能力均明显高于各浓度NaCl溶液处理组;蚊虫总感染率蒸馏水对照组均显著高于NaCl溶液处理组。     

Presystemic influences on thirst, salt appetite, and vasopressin secretion in the hypovolemic rat

The present studies investigated the influence of presystemic signals on the control of thirst, salt appetite, and vasopressin (VP) secretion in rats during nonhypotensive hypovolemia. Rats were injected with 30% polyethylene glycol (PEG) solution, deprived of food and water overnight, and then allowed to drink water, 0.15 M NaCl, or 0.30 M NaCl. The PEG treatment, which produced 30-40% plasma volume deficits, elicited rapid intakes in an initial bout of drinking, but rats consumed much more 0.15 M NaCl than water or 0.30 M NaCl. In considering why drinking stopped sooner when water or concentrated saline was ingested, it seemed relevant that little or no change in systemic plasma Na(+) concentration was observed during the initial bouts and that the partial repair of hypovolemia was comparable, regardless of which fluid was consumed. In rats that drank 0.15 M NaCl, gastric emptying was fastest and the combined volume of ingested fluid in the stomach and small intestine was largest. These and other observations are consistent with the hypothesis that fluid ingestion by hypovolemic rats is inhibited by distension of the stomach and proximal small intestine and that movement of dilute or concentrated fluid into the small intestine provides another presystemic signal that inhibits thirst or salt appetite, respectively. On the other hand, an early effect of water or saline consumption on VP secretion in PEG-treated rats was not observed, in contrast to recent findings in dehydrated rats. Thus the controls of fluid ingestion and VP secretion are similar but not identical during hypovolemia.     

The Mechanism of Isotonic Water Transport

The mechanism by which active solute transport causes water transport in isotonic proportions across epithelial membranes has been investigated. The principle of the experiments was to measure the osmolarity of the transported fluid when the osmolarity of the bathing solution was varied over an eightfold range by varying the NaCl concentration or by adding impermeant non-electrolytes. An in vitro preparation of rabbit gall bladder was suspended in moist oxygen without an outer bathing solution, and the pure transported fluid was collected as it dripped off the serosal surface. Under all conditions the transported fluid was found to approximate an NaCl solution isotonic to whatever bathing solution used. This finding means that the mechanism of isotonic water transport in the gall bladder is neither the double membrane effect nor co-diffusion but rather local osmosis. In other words, active NaCl transport maintains a locally high concentration of solute in some restricted space in the vicinity of the cell membrane, and water follows NaCl in response to this local osmotic gradient. An equation has been derived enabling one to calculate whether the passive water permeability of an organ is high enough to account for complete osmotic equilibration of actively transported solute. By application of this equation, water transport associated with active NaCl transport in the gall bladder cannot go through the channels for water flow under passive conditions, since these channels are grossly too impermeable. Furthermore, solute-linked water transport fails to produce the streaming potentials expected for water flow through these passive channels. Hence solute-linked water transport does not occur in the passive channels but instead involves special structures in the cell membrane, which remain to be identified.