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Additionally, chloral hydrate and pentobarbital sodium nembutal ; can produce profound sedation in the child with the risk of airway compromise and deep sedation. Chloride currents activated by ACh, SHT, glutamate, glycine, and GABA Gundersen et al., 1983a, 1984a, c; Kusano et al., 1982 ; . The reversal potential of the response to GABA was virtually unchanged in the presence of pentobarbital, even though larger currents were obtained at potentials away from the equilibrium Figs. 2B, 3 ; . The rectification at negative potentials was also still present Fig. 3 ; . Thus, the potentiation of the GABA response by pentobarbital seen at -60 mV does not arise from any shift in equilibrium potential or change in form of the current-voltage relationship. Instead, the degree of potentiation was similar at all potentials examined. Purpose: To seek behavioural, reflexive and histochemical evidence of long-lasting changes in nociceptive stimulus transmission induced by exposure to doses of pentobarbital that induce nocifensive hyperreflexia. Methods: Nocifensive hyperreflexia was induced in 12 rats with 30 mgkg1 pentobarbital ip. Reflex latency times for withdrawal of the hind paw from noxious radiant heat were measured with an automated electronic timer. Subjective responses to noxious stimulation licking or biting of the stimulated hindpaw ; and the level of sedation were recorded. Histological sections of lumbar spinal cord were stained for immunoreactivity of the immediateearly-gene IEG ; , c-fos, in three rats that received repeated threshold noxious radiant heat stimulation during the period of nocifensive hyperreflexia induced by 30 mgkg 1 pentobarbital ip. Results: Reflex withdrawal latency decreased by 32 8% of control values P 0.001 ; following pentobarbital injection and returned to control values 120 min after drug injection. Once fully alert, pentobarbital-treated animals did not show any increase in nociceptive behaviour relative to saline-injected controls P 0.41 ; . Sustained noxious stimulation to the hindpaw in halothane-anesthetized animals was associated with an increase in c-fos immunoreactivity in the dorsal horn of the lumbar spinal cord ipsilateral to the stimulation P 0.001 ; . Threshold stimulation in the pentobarbital-treated animals was not associated with any increase in c-fos expression. Conclusions: During pentobarbital-induced hyperreflexia, rats did not show any reflexive, behavioural, or histochemical evidence of long-lasting enhancement of nocifensive signal transmission. The results are consistent with previous observations that, in the absence of tissue injury, nocifensive hyperreflexia induced by barbiturates is a short-lived pharmacological effect. Objectif : Dcouvrir les manifestations comportementales, rflexes et histochimiques de modifications persistantes de la transmission d'un stimulus nociceptif induit par l'exposition des doses de pentobarbital qui provoquent une surrflectivit dfensive. Mthode : La surrflectivit dfensive a t induite chez 12 rats avec 30 mgkg1 de pentobarbital ip. Les temps de latence rflexe ncessaire au retrait de la patte arrire d'une source de chaleur radiante ont t mesurs avec un chronomtre lectronique automatis. Les rponses subjectives la stimulation dsagrable lcher ou mordre la patte stimule ; et le niveau de sdation ont t enregistrs. Des sections histologiques de la moelle pinire lombaire ont t colores pour vrifier l'immunoractivit du gne prcoce immdiat GPI ; , c-fos, chez trois rats qui ont reu une stimulation liminale nocive rpte de chaleur radiante pendant la priode de surrflectivit dfensive induite par les 30 mgkg1 de pentobarbital ip. Rsultats : Le temps de latence rflexe a baiss de 32 8 % par rapport aux valeurs tmoins P 0, 001 ; aprs l'injection de pentobarbital et est revenu aux valeurs tmoins 120 min aprs l'injection du mdicament. Une fois compltement rveills, les animaux traits au pentobarbital n'ont pas affich de comportement nociceptif accru compars aux animaux tmoins qui on a inject une solution sale P 0, 41 ; . stimulation nocive la patte arrire, subie par les animaux anesthsis l'halothane, a t associe avec un accroissement de l'immunoractivit au gne c-fos dans la corne suprieure de la moelle pinire lombaire homolatrale la stimulation P 0, 001 ; . La stimulation liminale chez les animaux traits au pentobarbital n'tait pas accompagne d'une augmentation de l'expression de c-fos . Conclusion : Pendant la surrflectivit lie au pentobarbital, les rats n'ont pas donn de signe rflexe, comportemental ou histochimique d'une augmentation persistante de la transmission du signal dfensif. Ces rsultats confirment des observations antrieures qui montraient qu'en l'absence de lsion aux tissus la surrflectivit dfensive induite par les barbituriques prsente un effet pharmacologique de courte dure.

As part of a program of research designed to investigate the long-term effects of early feeding experiences, the present study exploited the substantial flavor variation inherent in three classes of commercially available infant formulas: traditional milk-based formulas; formulas based on soy proteins; and those based on hydrolysed proteins. To this aim, we evaluated the preferences of 4to 6-year-old children n 102 ; who were divided into three groups based on the type of formula they were fed as infants. Children played a `taste and smell' game with a wide range of food-related odor qualities including infant formulas, as well as the flavor of milk-based and hydrolysate formulas and differently flavored apple juices. The data revealed that the type of formula that children were fed during infancy influenced their preferences when tested several years later. Children who were fed protein hydrolysate formulas were more likely to prefer the sour-flavored juices, as well as the odor and flavor of formulas, and were less likely to make negative facial responses during the taste tests when compared to children who were fed milk-based formulas. That the effects of differential formula feeding also modified children's food preferences is suggested by the mothers' reports that those fed hydrolysates were significantly more likely to prefer broccoli than were those fed milk formulas. These data are consistent with the. And spleen were only tested in animals from the F1 generation. In each experiment, nontransgenic littermates were used as controls. Because siblings have a very similar genetic background, the utilization of nontransgenic littermates as controls significantly enhances the precision of the observations. Analysis of transgene expression. Tissue concentrations of NPY were measured using a specific antiserum and radioimmunoassays as previously described 5, 25, 26 ; . For dissection of tissues, rats were anesthetized with pentobarbital sodium injection and perfused with saline via the left cardiac ventricle. For dissection of the hypothalamic areas, the brains were quickly removed, frozen on dry ice in isopentane, and stored at 80C. Serial sections of 300 m were cut, and discrete brain areas were microdissected. The landmarks for this dissection were derived from an atlas of the rat brain 34 ; . Total protein content in brain samples was determined by the bicinchoninic acid protein assay Pierce, Rockford, IL ; . NPY distribution in the coronal sections of the brain was determined by immunocytochemistry using the indirect peroxidase-antiperoxidase technique as described in detail 39 ; . Blood samples were collected from conscious rats using chronic vascular catheters dispensing to ice-cold tubes containing 0.5 trypsin-inhibitory unit of aprotinin and 10 IU of heparin. Plasma was immediately separated by centrifugation 14, 000 g, 4C, 15 min ; and stored at 70C until assayed. This protocol has been reported to provide plateletfree plasma and to prevent NPY release from the platelets 46 ; . Direct blood pressure measurement. Direct systolic and diastolic arterial blood pressures and heart rates were measured in resting, unrestrained, and conscious rats using a chronic arterial catheter. For implantation of a catheter, the rat was anesthetized with pentobarbital sodium 50 mg kg ip ; . With the use of a sterile technique, polyurethane tubing 0.033 in. OD and 0.014 in. ID, Micro-Renathane, Braintree Scientific, Braintree, MA ; was inserted into the lower abdominal aorta through the femoral artery. At the same time, another catheter was inserted into the inferior vena cava via the femoral vein for the administration of drugs. Both catheters were then exteriorized at the back of the neck, filled with a mixture of heparin 100 U ml ; , dextrose 50% ; , and penicillin G 1, 000 U ml ; in saline, secured with a silk suture, and plugged with a piece of fish line. During the 3- to 4-day recovery period, the animals were housed individually with food and water ad libitum. To adapt the animals to the and pentostatin.

Yield a final concentration of 106 cells ml. The cul- teers from the foreleg veins at 0.5, 1, 2, and 6 h. tures were then incubated at 37 C with shaking for The concentrations of antibiotic in each serum saman initial 8 h and without shaking for a subsequent ple was determined by the disk method, using 16-h period. The number of viable cells was mea- standard solutions prepared with sera of respective sured at regular intervals throughout the incuba- animals and volunteers. ii ; Urinary excretion. Urine of rats was coltion period. Protein binding. i ; Ultrafiltration method. To lected with a urine collector at 0 to 3, and 6 to 4.5 ml of human serum Consera ; or of fresh serum 24 h after intramuscular injection of 20 mg of the from each animal, 0.5 ml of antibiotic solution 300 test drugs per kg. Urine was collected from dogs and gg ml ; in phosphate buffer pH 7.0 ; was added monkeys with a catheter at specified intervals after and incubated at 37 C for 1 h. The mixture was intramuscular injection. The concentration of antipoured into a Visking tube 8 32 in size ; hung in a biotic in each urine sample was bioassayed with the 15-ml polypropylene tube and centrifuged at 1, 000 x standard buffer at pH 7.0, and the urinary recovery g for 30 min. The concentration-free antibiotic in the rate was calculated. iii ; Identification of active substances excreted ultrafiltrate was determined by the disk method. When these values are expressed as X and the val- into urine. Ceftezole was given intramuscularly to ues obtained in the reference experiment with buffer experimental animals 20 mg kg ; and human volunin place of serum are expressed as Y, the binding teers 500 mg ; . The urine was collected over a period rate B ; ofthe antibiotics was determined as follows: of 6 h after administration and examined by thinlayer chromatography and bioautography. The standard ceftezole solution 1 mg ml ; and urine samples x 100 B ; y were examined chromatographically by the use of Eastman Chromagram sheet no. 6061. The sheet was ii ; Effect of buffer dilution on antibiotic-protein then dried and developed with the solvent system nbinding. The antibiotic solution 500 , Ag ml ; was butanol-acetic acid-water 4: 1: 5, top layer ; . The dried mixed with ninefold volumes of human serum Con- sheet was then placed on an agar plate that had sera ; . The mixture was allowed to stand at room been seeded with 0.2% of the spore suspension 2 x temperature for 1 h, and then was diluted two-, 108 spores ml ; of B. subtilis ATCC-6633. iv ; Biliary excretion. Rats anesthetized with four-, and eightfold with M 15 phosphate buffer pH 7.0 ; . The concentration of antibiotic in each pentobarbital were fixed in a supine position, and a dilution was determined by the disk method. When polyethylene cannula was inserted into the bile M 15 phosphate buffer was used instead of serum, duct. Bile samples were collected at 0 to 3, and the concentration of antibiotic was expressed in 6 to after intramuscular injection of 20 mg of the test drugs per kg. The antibiotic levels in the bile terms of relative value as 100. Protective effect on infections in mice. Male ICR samples were assayed with the standard solutions white mice, 4 weeks of age and each weighing 17 to prepared with M 15 phosphate buffer pH 7.0 ; . v ; Tissue distribution. Groups of three rats each 23 g, were used in groups of 10 mice each. The organisms were cultivated overnight at 37 C received intramuscularly 20 mg of each of the test brain heart infusion agar and were then suspended antibiotics per kg and were sacrificed at 30, 60, and in 2 or 5% mucin solution to obtain microbial cell 90 min after drug administration. Liver, kidneys, concentrations of 1 x 106 to 4 x 106 ml. Mice were lungs, heart, and spleen were removed and after inoculated intraperitoneally with 0.5 ml of the sus- light washing in saline solution, the organs from pension. Each of the test antibiotics was adminis- each group of animals were pooled, mixed with tered subcutaneously in varying dosages to a group ethanol, and homogenized in a polytron homogeof 10 mice, 1 h after challenge. The mean effective nizer. The concentrations of antibiotic in the superdose ED50 ; values were found by the probit method natants obtained by centrifuging the tissue homogefrom the number of surviving animals after 2 weeks nates at 10, 000 rpm for 10 min were bioassayed with the standard solutions prepared with M 15 phosof observation. Microbiological assay. Each 10 ml of agar me- phate buffer pH 7.0 ; containing 66% ethanol. Each dium 1% sodium citrate, 0.5% polypeptone, 0.3% experiment was repeated three times in the same meat extract, and 1% agar ; was inoculated with 105 manner, and the values were averaged. vi ; Antibiotic concentrations in pouch exudates. spores of Bacillus subtilis ATCC 6633 per ml and placed in petri dishes. Test disks 6 or 8 diam- After subcutaneous injection of 20 ml air into the eter were immersed in the standard solutions or test backs of rats, 1 ml of 1% olive oil containing croton solutions containing the antibiotics under study. oil was injected into the pouch of each animal to After removing excess water, the disks were placed obtain aseptic inflammation. A 20-mg amount of on the above media. After standing at 37 C for 20 h, each antibiotic per kg was given intramuscularly on the diameters of the inhibitory zones were measured. day 6 or 7 after formation of the pouch, and the Absorption and excretion. i ; Drug concentra- exudates were collected at 0.5, 1, 2, and 4 h. The tions in serum. The test drugs were given intramus- antibiotic concentrations in the exudates were cularly in a dose of 20 mg kg to experimental ani- bioassayed with the standard solutions diluted with mals and in a dose of 500 mg to healthy human the exudates. volunteers. Groups of 10 rats each were anestheRESULTS tized with chloroform, and blood was collected from Antimicrobial activity. In the experiments the heart at 0.25, 0.5, 1, and 2 h. The blood was collected from dogs, monkeys, and human volun- conducted as a part of this study, ceftezole and peppermint.

National Chemotherapy Service Center, National Institutes of Health, Bethesda, Md. ; , vincristine Eli Lilly & Co., Indianapolis, Ind. ; , and Salmonella typhosa 0901 W lipopolysaccharide Difco ; were dissolved or suspended in sterile 0.15 M NaCl and adjusted to pH 7.0. Cyclophosphamide Mead Johnson Laboratories, Evansville, Ind. ; and methotrexate Lederle Laboratories, Pearl River, N.Y. ; were dissolved in sterile distilled water and adjusted to pH 7.0. Poly I: C was supplied as a stock solution of 1 mg per ml in 0.01 M phosphate-buffered 0.15 M NaCl pH 7.6 to 7.8 ; . Use of this stock solution necessitated ip injections of 0.02 ml per g of mouse weight 20 mg of poly I: C per kg ; . All drug plus endotoxin injections were performed simultaneously unless indicated otherwise. Probit analyses of data leading to LDw determinations, tests for parallelism, and calculation of potency ratios were done by the method of Litchfield and Wilcoxon 9 ; . The strain of P. aeruginosa employed in these studies was isolated from BALB c mouse feces. The culture was maintained on Nutrient Agar Difco ; . Cultures grown in peptone-yeast extract 0.5 and 0.3%, respectively ; broth for 18 hr at were harvested by centrifugation and suspended in sterile saline. The cell suspensions were heated in a water bath at 58 C for 1 hr. The number of viable organisms present in the suspensions before and after heating was determined as colony-forming units after 24 hr of incubation at 37 C Nutrient Agar or Penassay Agar Difco ; . The LD5o for mice, determined 3 days after injection, was 7 X 108 cells per kg. Pretreatment of mice with S. typhosa endotoxin involved the following regimen: 2 mg of endotoxin per kg on day -5, 4 mg of endotoxin per kg on days -4 and -3, and 8 mg of endotoxin per kg on day -2. The mice were challenged on day 0. Sedation of BALB c mice was measured after ip injections of aqueous solutions of 45 mg of sodium pentobarbital per kg Diamond Laboratories, Des Moines, Iowa ; . The duration of pentobarbital-induced sedation was expressed as the time interval between the loss and restoration of the righting reflex ability or disability to right themselves two times within a 10-sec period ; . Differences in the duration of sedation between test mice and control mice were evaluated by "one-tailed" Student t tests at the P 0.01 level of and percodan.

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