Use of a protective solution in the inoculation of bacterial cultures

The influence of the composition of solutions used for growing the bacterial cultures on the survival of microorganisms has been studied. The use of a protective solution, consisting of sodium chloride, orthophosphates, magnium sulfate, and gelatin, for suspensions of bacterial cultures has provided a higher isolation rate of enteric microorganisms, vs. cultivation with 0.9% sodium chloride solution. In evaluating the sensitivity of thioglycol medium test cultures are recommended to be inoculated with the above protective solution.     

Microbially initiated pitting on 316L stainless steel

Pitting corrosion of 316L stainless steel ennobled in the presence of manganese-oxidizing bacteria, Leptothrix discophora, was studied in a low-concentration sodium chloride solution. Corrosion coupons were first exposed to the microorganisms in a batch reactor until ennoblement occurred, then sodium chloride was added, which initiated pitting. The pits had aspect ratios (length divided by width) and shapes closely resembling the aspect ratio and the shape of the bacteria, which suggested that the microorganisms were involved in pit initiation.     

Development of a Simple Culture Method for the Tissues Contaminated with Microorganisms and Application to Establishment of a Fish Cell line

Cell cultures are good in vitro model systems in place of using living animals. In the present study, we developed a simple culture method in which tissues were pretreated with a low concentration of sodium hypochlorite solution (NaClO) to prevent not only bacteria but also fungi. Scales removed from a goldfish (Carassius auratus) body were treated with 70% ethanol and then with 0.3% of sodium hypochlorite solution, and cultured in vitro in an atmosphere containing 0.5% CO2. The doubling time of the established cells (GAKS) was 24 hr. The GAKS cells contained alkaline phosphatase activity (8.3+/-1.1 nmol/min/mg protein) and secreted 0.32+/-0.07 pg/ml endothelin during a 3 day culture of a full monolayer sheet.     

RESPIRATION AND MITOCHONDRIAL CONTENT IN SINGLE NEURONS OF THE SUPRAOPTIC NUCLEUS : A Correlative Study in Osmotic Stress

The study was undertaken to investigate the possible correlation of total volume of mitochondria per cell with the rate of succinate oxidation in isolated nerve cell bodies, after various functional stresses in the experimental animals. Significant cytological effects were found in the nerve cells of the supraoptic nucleus in rats which had been thirsting for 4–12 days or had been given 2% sodium chloride solution as a substitute for drinking water for a few weeks. Quantitation of mitochondria was done from electron micrographs. The cell volumes were calculated from sections of Epon-embedded tissue under phase-contrast microscopy. Succinate oxidation was measured on groups of 10 nerve cells with the microdiver technique. As a result of either thirst or sodium chloride load, the volume of mitochondria per nerve cell more than doubled. The rate of succinate oxidation was not changed after the rats had been thirsting but was enhanced by over 100% after they had drunk sodium chloride. A linear relationship was found for the amount of mitochondria versus respiration in the supraoptic neurons for all experimental groups except the thirsting animals. The mitochondria in the supraoptic neurons from thirsting animals were of the same size or smaller than those in controls, whereas in animals given sodium chloride solution the mitochondria were considerably enlarged. The observed effects were specific for the supraoptic nucleus.     

Intake of water and sodium chloride solution by the rats after central administration of litorin and immunization with litorin-bovine serum albumin conjugate

Central injection of litorin suppressed water and sodium chloride solution intake, and also decreased the preference of saline of unrestrained rats during 1 hour test. Active immunization of animals with litorin-bovine serum albumin conjugate led to the augmentation of their daily saline preference. Immunized rats increased also avidity to salt after deprivation, though the consumption volume of liquids decreased. The data, obtained on the immunized animals prove the importance of definite level of endogenous bombesin-like peptide in the salt appetite of rats. The central effect of litorin is discussed in the aspect of changing by it the evaluation of gustatory information.     

Paddle cilia and discocilia — Genuine structures?

Kinocilia of epidermal sensory cells in fixed marine Turbellaria often terminate as flattened biconcave discs. The distal part of the ciliary axoneme curves back upon itself forming a 360 degree loop which is enveloped by the plasmalemma. In living animals this structure can be induced by the addition of sodium cacodylate, monobasic sodium phosphate, dibasic sodium phosphate, sucrose, calcium chloride, or formaldehyde to the sea water. Specimens treated with sodium chloride, glutaraldehyde, or osmium tetroxide do not show modified cilia. In animals prepared for EM at low temperature and with a buffered hypotonic fixative less kinocilia are modified than in animals treated with a buffered iso- or hypertonic fixative and at a higher temperature. It is assumed that the unusually shaped cilia, described as "paddle cilia" or "discocilia" in other invertebrates, do not represent a genuine but an artificial structure.     

Factors associated with Pseudomonas pickettii intrinsic contamination of commercial respiratory therapy solutions marketed as sterile.

Laboratory investigations were conducted to study the growth dynamics of Pseudomonas pickettii in commercial 0.9% sodium chloride solution under various environmental conditions and to determine the retention of these organisms after challenge through a 0.2-micron cartridge filter system. Low numbers of P. pickettii (1 to 10 CFU/ml of test solution) inoculated into commercial vials containing 5 ml of 0.9% sodium chloride solution and 500-ml volumes of 0.9% sodium chloride solution were shown to proliferate over a 168-h incubation period. These organisms demonstrated growth over a wide range of temperatures (15 to 42 degrees C) in this salt solution, and survival studies at 50, 55, and 60 degrees C indicated that this strain was not unusually resistant to heat (with the times required at a given temperature to reduce the surviving microbial population 10-fold [D-values] being 26.0, 1.9, and 0.7 min, respectively). A challenge test demonstrated that P. pickettii organisms were not completely retained by a 0.2-micron cartridge filter. The number of organisms detected increased from 1 CFU/liter of effluent at 1 to 2 min to a maximum of 176 CFU/liter at 4 to 5 min. Our results indicate that P. pickettii can penetrate a 0.2-micron filtration system and that the passage of organisms and subsequent microbial growth in the filter effluent probably are the mechanisms by which these organisms were recovered from "sterile" commercial 0.9% sodium chloride solution.     

Studies on Bacterial Pyrogenicity: III. Specificity of the United States Pharmacopeia Rabbit Pyrogen Test

The specificity of the first or “presumptive” portion of the USP rabbit pyrogen test was investigated by use of a new absolute standard of reference. The reference standard was a 0.9% sodium chloride solution prepared to be pyrogen-free. Details of the preparation were described. The hypothesis was explored that the temperature response of rabbits after intravenous injection of the standard solution was independent of exogenous pyrogen. Reactions observed among the rabbits in our colony allowed a classification of these animals ranging from “consistently reliable” to “consistently unreliable.” Details of the experimental results and implications for pyrogen testing are discussed. The recommendation was made that all rabbit test animals be “screened” in sham and actual tests before being used for pyrogen testing.     

Loss of basophilic (sulphated) material from sections of cartilage treated with periodate solution

Synopsis Fresh-frozen sections of cartilage quickly lost much basophilic material (presumably chondroitin sulphate) when placed in a 2% aqueous solution of sodium periodate. The loss is due specifically to the periodate anion. Hydrogen peroxide was as active as sodium periodate in this respect, but little loss occurred in the presence of various thiols.Proteoglycan and connective tissue polysaccharide showed rapid and marked decreases in viscosity on oxidation in low concentrations of aqueous periodate, although only limited oxidation of glycols had taken place.Pretreatment of sections with cetylpyridinium chloride solution greatly diminished the loss of basophilia as did the incorporation of high concentrations (>15%) ofn-propanol in the periodate solution. Acid solutions (pH 3.0) during oxidation, or as a pretreatment, also reduced the loss. This fixation could not be reversed by exposure to neutral buffer or concentrated salt solutions.It is suggested that the loss of basophilia from sections treated with sodium periodate solutions is due to the degradation of proteoglycan, possibly by a free radical mechanism. The relevance of the loss to the structure of cartilage is considered.     

The observation of partial regeneration of the tip of tail in newborn rat pups in a liquid medium

When the wound surface formed as a result of amputation of the tail tip in newborn rats was placed in sterile 0.9% aqueous solution of sodium chloride until full epithelialization of the defect, the epithelium moved onto the blood clot covering the wound. The blood clot was then substituted for connective tissue, into which the traumatized vertebra protruded. The vertebra restored its anatomical integrity, and this led to partial regrowth of the tail. A skin regenerate was formed on the apical tail surface with characteristic features of the intact skin of this locality. In the control animals, no regeneration of the vertebra epiphysis took place and a scar was formed at the stump end.     

Droplet freezing of antibody-linked indicator red cells of sheep, ox, and human origin

A droplet freezing technique for the cryopreservation of indicator red cells is described. Recovery was crucially dependent on the composition of the solution in which the cells were suspended. Preliminary experiments to determine the relative importance of sucrose, glucose, sodium chloride and hydroxyethyl starch (HES) in determining the survival of trypsin-treated sheep red cells showed that the addition of sucrose or HES or both to isotonic sodium chloride solution increased recovery, whereas the additional inclusion of glucose was detrimental. It was shown that glucose penetrated the cells whereas sucrose did not. The optimum combination of sucrose and sodium chloride concentration, in the presence of 6 g/dl HES, was 7 g/dl sucrose plus 0.3 g/dl sodium chloride. Recovery was increased by increasing the concentration of HES, and maximal recovery was obtained by thawing the frozen droplets in phosphate-buffered saline at 40 °C. Trypsintreated ox and human cells gave much lower recovery than sheep cells when HES was used in the freezing mixture but the substitution of dextran (10 g/dl) for HES gave greater than 80% recovery with all three species. Ten different antibody-coupled reagent cells all gave >83% recovery. The effects of hematocrit, incubation time, and storage temperature are described. The preservation technique described is simple and convenient, and will make it possible to extend the use of immunoassay procedures using antibody-coupled red cells.     

Natriuretic reaction of the vertebrate kidney

After injection of a 10% solution of NaCl, the kidney of rats, and to a lower extent that of hens, rapidly excretes the excess sodium. Effective sodium excretion in homoiothermic animals is based on high level of glomerular filtration and proximal reabsorption. After injection of a 10% solution of NaCl to the Lamprey Lampetr fluviatilis and frog Rana temporaria, sodiumuretic reaction of the kidney was less evident. During water diuresis, cells of renal tubuli of lampreys, frogs, hens and rats produce high concentration gradient between the blood serum and urine, preventing loss of sodium. In evolution, the kidney was formed presumably as an organ which prevents sodium loss from the organism; this function remains highly effective in all vertebrates, whereas in homoiothermic animals the kidney attains also the capacity for rapid extrusion of excessive sodium.     

高渗氯化钠联合纳洛酮对失血性休克患者的临床疗效及安全性分析

目的:探讨高渗氯化钠联合纳洛酮对失血性休克患者的临床疗效及安全性。方法:收集我院收治的72例创伤失血性休克患者,随机分为实验组和对照组,每组36例,两组入院后均给予ICU常规治疗,对照组患者给予纳洛酮注射液1.2 mg加入5%葡萄糖静注,5 min/次,3次后改用4 mg持续静滴;实验组患者在对照组的基础上给予10%氯化钠液220 m L加入生理盐水80 m L配成7.5%的高渗氯化钠溶液静脉滴注。治疗结束后,对两组患者苏醒时间、血清NO、CD18水平以及临床疗效进行检测和比较。结果:与治疗前相比,两组患者的血清NO、CD18水平均下降(P0.05);与对照组相比,实验组患者的苏醒时间较短,血清NO、CD18水平较低,并发症的发生率也较低(P0.05)。结论:高渗氯化钠联合纳洛酮能够提高失血性休克患者的临床疗效,且安全性高,可能与其降低失血性休克患者血清NO、CD18水平有关。     

Effects of endotoxic shock on diaphragmatic function in mechanically ventilated rats.

Diaphragmatic function was investigated in mechanically ventilated rats during endotoxic shock (group E, n = 18) and after saline solution injection (group C, n = 8). Endotoxic shock was produced by a 1-min injection of Escherichia coli endotoxin (10 mg/kg iv) suspended in saline. Diaphragmatic strength was assessed before (T0) and 15 (T15) and 60 (T60) min after injection by measuring transdiaphragmatic pressure (Pdi) generated during bilateral phrenic stimulation at 0.5, 10, 20, 30, 50, and 100 Hz. Diaphragmatic neuromuscular transmission was assessed by measuring the integrated electrical activity of the diaphragm. Diaphragmatic endurance was assessed 75 min after injection from the rate of Pdi decline after a 30-s continuous 10-Hz phrenic stimulation. In 16 additional animals, diaphragmatic glycogen content was determined 60 min after inoculation with endotoxin (n = 8) or 0.9% sodium chloride solution (n = 8). Diaphragmatic resting membrane potential (Em) was measured in 16 additional animals 60 min after endotoxin (n = 8) or saline injection (n = 8). Mean blood pressure decreased from 74 +/- 3 to 53 +/- 6 mmHg at T60 in group E, whereas it was maintained in group C. At T60 Pdi was decreased in group E for frequencies of 50 and 100 Hz and was associated with a decreased diaphragmatic electromyographic activity of 25.3 +/- 2.5 and 26.5 +/- 5.2% for 50- and 100-Hz stimulations, respectively, in comparison with T0 values.(ABSTRACT TRUNCATED AT 250 WORDS)     

Combined effect of alkali pretreatment and sodium chloride addition on the olive fermentation process

Green olives of the Tunisian variety "Meski" were treated according to a Spanish-style green olive preservation process by using an alkaline treatment (1.5, 2 and 2.5% (w/v) NaOH) to eliminate bitterness, combined with different brine concentrations (6, 9 and 12% (w/v) NaCl). A spontaneous fermentation by the environmental microflora took place. Results showed that 2% NaOH solution and 9% sodium chloride brine was an optimal combination inducing the best growth of Lactobacillus species (10(8) CFU/ml) and acidity of 0.726 g lactic acid/100 ml brine. In all trials and independently of the treatment, Lb. plantarum was the most dominant strain of Lactobacillus. Moreover, pretreatment with lye and lactic fermentation of olives contributed to coliform elimination.     

An automatic method for the measurement of the effect of short stresses on the microbial ATP pool

An automatic method for measuring the effects of brief contacts between microorganisms and toxic agents is described.In this method, both a microbial culture and a toxic solution are injected into a mixing tube, until inhibition of bacteria and ATP extraction take place.The range of the contact times, defined as the residence periods of the cultures in the mixing tube, is quite wide (from 0.3 to 16 s), with increased coefficient of variation (from 5 to 14%) for the lower times.     

Specific changes in the sodium chloride intake and activity of the taste receptor apparatus of rats administered the peptide litorin and its albumin conjugate

The role of bombesin-like peptide litorin in control of salt consumption was studied in rats. The influence of exogenous injection and binding of the endogenous litorin in the organism on the consumption of sodium chloride and sucrose solutions was analyzed in deprived rats in the process of 15-min. testing. It was found that intraperitoneal injection of 10(-6) - 10(-5) M of litorin led to an increase, and rats immunization by peptide conjugate with ox serum albumin--to a decrease of the consumption of salt solution, without changing the consumption of sucrose solution. Titres of antibodies to litorin in the plasma of rats immunized with the conjugate were higher than in the control. Intraperitoneal, subcutaneous or intranasal injections of 10(-5) M of litorin led to selective increase of gustatory responses (recorded from the chorda tympani) to tongue stimulation by salt and acid solutions not influencing the responses to quinine and sucrose in anesthetized rats. The study shows, that the peptide litorin selectively participates in the control of sodium chloride consumption. In the process of such control the increase of taste apparatus sensitivity to the sodium chloride is probably involved.     

Effect of lithium preparations on the toxic effects of adrenaline

Experiments on rats have shown that intravenous injection of adrenaline in a dose of 0.3-0.4 mg/kg causes cardiac arrhythmia. In this case the primary arrhythmia developing immediately after adrenaline injection is followed by the recovery of sinusal rhythm which was replaced by the secondary arrhythmia. Apart from arrhythmias, there developed pulmonary edema. The animals died 2--3 minutes after adrenaline injections. Lithium chloride and lithium hydroxybutyrate removed the secondary arrhythmia and pulmonary edema. Lithium hydroxybutyrate has proved to be more effective.     

Structure and Function of the Chloride Cell of Fundulus heteroclitus and Other Teleosts

Teleosts, the bony fishes, inhabit both freshwater and seawaterenvironments. Some euryhaline fish, such as Fundulus heteroclitus,alternate between the two milieux several times daily. Regardlessof adaptation, the gills of these animals possess a highly specializedcell type called the chloride cell. This cell contains numerousmitochondria and exhibits a greatly amplified basolateral cellsurface richly endowed with Na,K-ATPase. Recent studies on isolatedopercular epithelia containing chloride cells have demonstratedactive chloride secretion and passive transepithelial sodiummovements, and have established the chloride secretory roleof this cell type in seawater-adapted teleosts. Current modelssuggest that chloride transport occurs via a transcellular route.Seawater chloride cells exist in multicellular units and sharesimple, shallow tight junctions which are thought to be theroute for passive sodium movement. Freshwater chloride cells,whose function remains to be elucidated, are generally describedas existing in a unicellular configuration. However, recentobservations in Fundulus heteroclitus adapted to salinitiesas low as 1% sea water reveal that chloride cells persist inmulticellular complexes with apical crypts. Strikingly, tightjunctions between chloride cells in this freshwater environmentare deep     

Comparison of the instrumental reinforcer devaluation effect in two strains of rats (Wistar and Lister)

In this experiment, the effect of the reinforcer devaluation upon instrumental performance was analysed in two strains of rats (Wistar and Lister): Food deprived rats were trained to press a lever for sucrose pellets in a single session. Immediately after the fulfilment of this session, half of the Wistar and Lister rats received an injection of lithium chloride (LiCl), while the remaining animals were injected with a sodium chloride (NaCl) solution. A subsequent extinction test showed that the subjects who had received immediate LiCl did not press the lever as often as those injected with NaCl. No differences in response suppression were found between the two strains of rats. These results also show that a single devaluation experience is sufficient for an impact on instrumental performance.