Fourth Experimental Group Research Articles

Effect of selected feed additives on internal quality parameters of table eggs

The aim of our experiment was to evaluate under experimental conditions the influence of probiotic preparation based on lactobacillus, oregano essential oil, sumac (Rhus coriaria), propolis and pollen on egg quality parameters of laying hens hybrid Lohman Brown. For housing hens (n ​​= 60) three storey enriched battery cage was used in which hens were divided in groups (n = 10). Total floor space given to one animal was 943.2 cm2. In the control group of hens complete feed mixtures without any additions were fed. In the first experimental group complete feed mixture was enriched with probiotic lactobacilli based preparation in a dose of 0.5 g.kg-1. In the second experimental group oregano essential oil was added to the feed mixture at a dose of 0.5 ml.kg-1. The third experimental group was enriched with 1 % concentration of sumac (Rhus coriaria). The fourth experimental group was enriched with 0.2 g of propolis extract per 1 kg of feed mixture and the fifth experimental group was supplemented by pollen extract of the same dose. All groups were fed ad libitum. Eggs quality indicators observed were egg weight (g), yolk percentage (%), yolk index, white percentage (%), whites index and Haugh units of whites (HJ). The results show that addition of probiotics positively, although not significantly, influenced the average egg weight. The addition of oregano oil and sumac insignificantly decreased egg weight (P>0.05), the values ​​of the other eggs quality indicators were comparable with the control group. Supplementation of feed mixture with propolis as well as phytobiotics insignificantly decreased egg weight, but its addition improved the internal quality parameters as the yolks and whites index, whose average values ​​were in this group, although not statistically significantly higher (P>0.05).

Abstract B110: Chemoprevention of rat oral carcinogenesis by licofelone, a dual cyclooxygenase/lipoxygenase inhibitor

Abstract Cyclooxygenase [COX] inhibitors [piroxicam, celecoxib, naproxen, and NO-naproxen] are potent inhibitors of oral carcinogenesis induced in rats by 4-nitroquinoline-1-oxide [NQO]. By contrast, a model 5-lipoxygenase [LOX] inhibitor [zileuton] has no chemopreventive activity in the rat NQO oral cancer model. Bifunctional agents such as licofelone [6-(4-chlorophenyl)-2,2-dimethyl-7-phenyl-2,3-dihydro-1H-pyrrolizin-5-yl]acetic acid] can inhibit both COX and LOX pathways of eicosanoid biosynthesis. As a result of these activities, dual COX/LOX inhibitors (a) may be less toxic than inhibitors of a single pathway of arachidonic acid metabolism, and (b) may prevent redirection of eicosanoid biosynthesis into parallel pathways that stimulate inflammation and carcinogenesis. A chemoprevention efficacy evaluation of dual COX/LOX inhibition was performed using licofelone as a model compound. Squamous cell carcinomas of the tongue were induced in male F344 rats by administration of NQO [20 ppm] in the drinking water for 10 weeks. Beginning two days after completion of NQO administration, groups of 30 rats received daily oral [gavage] administration of licofelone at 0 mg/kg/day [vehicle control], 37.5 mg/kg/day [low dose], or 75 mg/kg/day [high dose]. A fourth experimental group received oral [gavage] administration of 75 mg licofelone/kg beginning 6 weeks post-NQO [high dose, delayed intervention]. Licofelone or vehicle administration was continued until study termination at week 25. When administered beginning two days after NQO, licofelone conferred dose-related protection against oral carcinogenesis [55% and 43% incidences of oral cancer in low and high dose groups, respectively, versus 75% in controls; p < 0.05 for high dose group]. Delayed administration of the high dose of licofelone demonstrated the greatest chemopreventive activity [34% oral cancer incidence versus 75% incidence in controls; p < 0.01]. Licofelone may inhibit oral carcinogenesis by delaying cancer progression; rats receiving the high dose of licofelone starting either 2 days or 6 weeks post-NQO demonstrated (a) decreases in the incidence of the most invasive [+4] cancers [17% in both groups versus 54% in controls; p < 0.01] and (b) increases in the number of rats without invasive malignancies [normal epithelium or epithelial hyperplasia only; 37% and 41% incidence versus 11% in controls; p < 0.01 for both comparisons]. The chemopreventive activity of the low dose of licofelone was marginally significant [0.05 < p < 0.10 for both comparisons]. Licofelone also prevented the pre-terminal body weight loss commonly seen in animals with advanced oral cancer in this model. These data demonstrate that licofelone is an effective inhibitor of oral carcinogenesis induced in rats by NQO, and suggest that licofelone chemopreventive activity results from inhibition or delayed progression of preneoplastic lesions to invasive oral cancers. The chemopreventive activity of delayed administration of licofelone is particularly notable because clinical trials are most likely to involve individuals with preexisting oral lesions. Dual COX/LOX inhibitors may provide an approach to oral cancer prevention that is not limited by the toxicity induced by specific of inhibitors of either COX or LOX. [Supported by NCI-N01-CN-25113] Citation Format: David L. McCormick, Thomas L. Horn, William D. Johnson, Ronald A. Lubet, Vernon E. Steele. Chemoprevention of rat oral carcinogenesis by licofelone, a dual cyclooxygenase/ lipoxygenase inhibitor. [abstract]. In: Proceedings of the Eleventh Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2012 Oct 16-19; Anaheim, CA. Philadelphia (PA): AACR; Cancer Prev Res 2012;5(11 Suppl):Abstract nr B110.

In vitro and In vivo antimicrobial activity of propolis on the microbiota from gastrointestinal tract of chickens

The aim of this study was to examine the effect of propolis extracts on the microbial colonization of chicken gastrointestinal tract in vivo. The propolis was administered to both feed mixtures in various amounts except of the control group. The addition of 150 mg propolis to 1 kg of feed was included in the first experimental group, the addition of 450 mg.kg−1 in the second experimental group, the addition of 600 mg.kg−1 the third experimental group and 800 mg kg−1 in the fourth one. The highest count of faecal enterococci was found in the third group (8.6 cfu.g−1) where 600 mg of propolis to 1 kg was added to the feed mixture. The highest count of lactobacilli was detected in the fourth experimental group (8.83 cfu.g−1) where was 800 mg of propolis added to 1 kg of feed mixture and number of Enterobacteriaceae genera count was found in control group (8.73 cfu.g−1). With RTQ PCR detected species from the genus Enterococcus were: E. avium, E. casseliflavus, E cecorum, E. faecalis, E. faecium, E. gallinarum, E. hirae and E. malodoratus and from genus Lactobacillus were: Lactobacillus crispatus, L. acidophilus and L. salivarius. With MALDI TOF MS Biotyper from Enterobacteriaceae genera were identified Citrobacter braakii, Raoultella ornithinolytica, Serratia fonticola, Escherichia coli and Klebsiella oxytoca. Antimicrobial activities In vitro of six species of bacteria isolated from gastrointestinal tract of chickens were also tested. The best antimicrobial effect of Citrobacter braakii on ethanolic propolis extract in all concentrations were found.

INFLUENCE OF ω-3 FATTY ACIDS SUPPLEMENTED TO LAYING HENS' DIET ON LAYING PERFORMANCE

A group-control experiment for a period of 45 days was conducted in order to determine the effects of use of ω-3 fatty acids on the production and economic results of laying hens. A total of 1264 Lohmann Brown layers were randomly devided into 4 experimental treatments with 79 replicates (4 per cage, 316 hens per treatment). The first group was fed complete feed mixture for laying hens with standard raw material and chemical composition. The feed mixtures for treatments 2., 3. and 4. were supplemented with ω-3 fatty acids. The source of ω-3 fatty acids was a microalgae Schizochytrium spp., commercial product “DHA Gold” (Martek, USA) in quantities of 0.5, 0.7 and 1.0%, respectively. The use of ω-3 PUFA supplement in higher inclusion rates, had a positive influence on the laying intensity, significantly higher (p <0.01) in the experimental groups supplemented in the amount of 0.7 and 1.0%. Layers of all experimental groups fed supplemented feed mixtures had lower average egg weights (68.1, 68.7 and 68.4 g) compared to the control group (68.8 g). However, supplementation with higher inclusion rates of ω-3 product resulted in higher total egg mass during the experiment (743 and 764 kg) in the third and fourth experimental groups, compared to the control group (691.9 kg). Experimental groups with a higher inclusion of ω-3 PUFA (0.7 and 1.0%), achieved higher daily consumption of 0.16% and 0.51% in comparison to the control group, but the best feed conversion (2.38 and 2.32), while supplementation in the amount of 0.5% resulted in the weakest conversion in the experimental group 2 (2.63). The lowest feed cost per produced egg was achieved by the control group fed complete, unsupplemented feed. Using supplemented feed with addition of ω-3 PUFA – DHA above the 0.5%, a significant difference was observed regarding the productivity of laying hens.

Establishing presence and status of infection with bovine diarrhoea virus in cattle herds in Central Serbia: Possibilities for control

The objective of our investigations was to determine the presence and status of infection with the bovine viral diarrhoea virus (BVDV) and to identify persistently infected (PI) cattle in 4 herds (experimental groups): 2 with fattening heifers (10 and 13 animals) and 2 with dairy animals with offspring older than 4 months (23 and 16 animals). Two series of blood samples were taken in an interval of 4 weeks (even samples) from all the animals, from each experimental group. Separate blood serums from both groups of samples were examined using the indirect ELISA method (HerdChek*BVDV Ab test kit). The results obtained in our investigations indicate the presence of infection with the BVD virus in the experimental animals. Blood serum samples taken on the first day of investigation were seropositive in 7 experimental animals of the first group, in all (n=13) experimental animals of the second group, seropositive in one and suspect in 2 animals of the third experimental group, and seropositive in one and suspect in 3 experimental animals of the fourth group. Blood serum samples taken on the 28th day of the examinations were seropositive in one, thirteen, two, and three experimental animals of the first, second, third, and fourth experimental groups. Serum samples of animals (n=29) that were established as seronegative in both investigations (on the first and the 28th day) were examined for the possible presence of antigen to the BVD virus using the ELISA method (HerdChek*BVDV Ag/Serum Plus) with the objective of investigating the presence of PI animals. The presence of antigen to the BVD virus was not established in the examined cattle. On the grounds of the results of the investigations and data in literature, measures for controlling BVDV infections were recommended to animal owners. .

Investigation of bacteremia after toothbrushing in orthodontic patients

Objective: The aim of this study was to investigate the occurrence rate of bacteremia following toothbrushing with toothpastes composed of several antibacterial agents and compare the results with the conventional oral hygiene maintaining methods in orthodontic patients. Methods: This clinical study included 100 adult orthodontic patients who were divided into 4 groups. Each group comprised of 25 patients, wearing fixed orthodontic appliances. In the first group, bacteremia was assessed after toothbrushing without using any toothpaste. In the second group, a 0.2% chlorhexidine gluconate mouthrinse was used before brushing with no toothpaste. In the third group, subjects brushed with a commonly used toothpaste which did not include an additional antimicrobial agent. The fourth experimental group used toothpaste which included tea tree oil, clove oil, peppermint oil and bisabolol as antimicrobial elements. Pre- and post-brushing blood samples were obtained using a strict aseptic technique. All samples were microbiologically evaluated using blood culture bottles. Results: Toothbrushing in orthodontic patients yielded to an increase in the occurrence rate of bacteremia when using normal toothpaste or no toothpaste at all. Conclusions: The use of chlorhexidine mouthwash before toothbrushing, and brushing with antimicrobial toothpaste did not show a statistically significant difference in preventing the occurrence of bacteremia (p ＞ 0.05).

The Effect of Using Four Teaching Styles in Developing some skills in Basketball For Female Pupils at Secondary First Year

The research aims at revealing the effect of using four teaching styles in developing some skills in basketball at first year female pupils. The study is based on the experimental approach due to its suitability. The Society of the research were 88 first year female pupils at their intermediate stage in Ba’sheeq. They were divided into four groups each of which consisted of 22 pupils. Equivalence concerning the variables age, height and weight was achieved in addition to a number of elements of physical and some basic skills in basketball. The first experimental group used the inclusion style in teaching whereas the second experimental group used small homogenous groups. The third used the sample of small heterogeneous groups while the fourth experimental group used the command style. The execution of the educational program lasted 8 weeks with the average of two educational units per week. The time devoted for each educational unit was 45 minutes. The data were collected and tested statistically by using mean, standard deviation, simple correlation coefficient (Person), T-test for equal correlated samples, analysis variance in one direction and Duncan’s L.S.R. The analysis showed the following findings: 1. The inclusion style and the two samples of small groups styles were found more effective in developing some basic skills like passing , dribbling and shooting in basketball. 2. the command style were found developing in skill pass. 3. The sample of small homogeneous groups compared with the other educational styles was found more effective in developing the shooting skill.

Imipramine treatment ameliorates corticosterone-induced alterations in the effects of 5-HT 1A and 5-HT 4 receptor activation in the CA1 area of rat hippocampus

This study tested whether imipramine reverses adaptive modifications in the function of hippocampal 5-HT 1A and 5-HT 4 receptors induced by repetitive administration of corticosterone. Rats received corticosterone for 1 or 3 weeks or imipramine for 2 weeks. The fourth experimental group was treated with corticosterone for 3 weeks and additionally with imipramine, beginning on the eighth day of corticosterone administration. Hippocampal slices were prepared 48 h after the last drug administration. 5-HT 1A and 5-HT 4 receptor-mediated effects on CA1 population spike amplitude were measured. While repeated corticosterone attenuated the inhibitory effect of 5-HT 1A receptor activation by 8-OH-DPAT and enhanced the excitatory effect of 5-HT 4 receptor activation by zacopride, imipramine treatment of naïve rats resulted in opposite changes. In the corticosterone plus imipramine group, the effect of 8-OH-DPAT and zacopride were not different from control, indicating that corticosterone-induced adaptive changes in the reactivity of 5-HT 1A and 5-HT 4 receptors were reversed by imipramine treatment.

Acute and Chronic Exposition of Mice to Severe Hypoxia: The Role of Acclimatization against Skeletal Muscle Oxidative Stress

The role of acclimatization and the effect of persistent severe hypoxia (7000 m) were analyzed in mice soleus muscle with respect to oxidative stress (glutathione redox status) and damage markers (TBARS and SH protein groups), NAG and SOD activities and HSP70 expression. Forty mice were divided into one normobaric-normoxic control group and four hypobaric-hypoxic experimental groups (n = 8). One experimental group (1 D) was acutely exposed to a simulated altitude of 7000 m in a hypobaric chamber for 1 day. Another experimental group (ACCL + 1 D) was exposed to a 3 days acclimatization period plus 1 day of hypoxia exposure at 7000 m. The third experimental group (ACCL + 8 D) was exposed to the same acclimatization protocol, remaining 8 subsequent days at 7000 m. The fourth experimental group (8 D) was chronically exposed without acclimatization. ACCL + 1 D showed a significant decrease (p < 0.05) in oxidative stress and damage compared to the 1 D group. Concerning chronic severe hypoxia, acclimatization was truly vital, since 8 D animals died after 5 days of exposure. Oxidative stress and damage markers in ACCL + 8 D tended to gradually increase throughout the 8 days of the hypoxic period. Total SOD activity did not change in 1 D compared to control; however, it increased significantly (p < 0.05) in ACCL + 1 D and ACCL + 8 D. HSP70 expression followed the observed oxidative stress and damage pattern, suggesting a protective role against hypoxia-induced oxidative stress. The present study supports the hypothesis that acclimatization attenuates oxidative stress and damage induced by acute hypoxia, although a trend to a gradually increased oxidative deleterious effect in skeletal muscle seems to occur during persistent severe hypoxia even after a previous acclimatization period.

Effects of some pharmacological agents on the survival of unipedicled venous flaps: an experimental study.

Clinical and experimental studies have been conducted to improve the survival of venous flaps. As a result of these studies, although various survival mechanisms were raised, none obtained satisfactory information. Venous stasis, and the resultant venous thrombosis, is a factor that decreases the survival of venous flaps. In this study, we evaluated the effects of two antiinflammatory agents, etodolac and etofenamate, on the survival of unipedicled venous flaps. In this study, 35 male New Zealand white rabbits (3,500-4,000 g) (70 ears) were used. Perichondrocutaneous flaps, 3 x 4.5 cm in size, were designed and raised, keeping the central veins intact in the middle of venous flap. Central arteries and nerves were ligated and transected both proximally and distally, to prepare unipedicled venous flaps. A silicone sheet was placed between the cartilage tissue and flap, to prevent blood flow and revascularization beneath. The subjects were divided into seven groups, consisting of five rabbits (10 ears). In the negative control group (group I), the single vascular pedicle of venous flaps, central veins were ligated and flaps sutured into their own place as the composite graft. In the positive control group (group II), after venous flaps were prepared, normal saline, 0.2 mL, was given subcutaneously. In the first of five experimental groups (group III), unfractionated heparin (100 U/day) was given subcutaneously. In the second experimental group (group IV), etodolac (5 mg/kg/day) was given subcutaneously. In the third experimental group (group V), etophenamate (5 mg/kg/day) was given orally through a feeding tube. In the fourth experimental group (group VI), parnaparin (5 anti-Xa U/kg/day) was given subcutaneously. In the fifth experimental group (group VII), nadroparin (5 anti-Xa U/kg/day) was given subcutaneously, about 7 days postoperatively. At the eighth postoperative day, surviving areas of venous flaps were measured, and the results were evaluated by Kruskal-Wallis ANOVA and Mann-Whitney U-test (P < 0.05). Biopsies were also taken from the flaps for histological evaluation of border of necrotic tissue. Surviving areas of unipedicled venous flaps were larger in experimental groups than those in negative and positive control group (P < 0.05). However, comparison of the experimental groups demonstrated no statistically significant difference (P > 0.05). We concluded that all pharmacological agents used in the experimental groups succeeded in increasing the survival of unipedicled venous flaps. Survival of the unipedicled venous flap was higher in venous flaps than that of composite graft, clearly showing the importance of the venous pedicle.

Role of the cytoskeleton in stimulation of Na+ channels in A6 cells by changes in osmolality.

Permeable supports with A6 cell monolayers were mounted in an Ussing chamber and bilaterally bathed with Ringer solution at room temperature. Short-circuit current (Isc) was recorded continuously, and noise analysis revealed microscopic channel current characteristics. Our investigation focuses on the stimulation of apical Na+ entry caused by exposing the serosal surface of the A6 cell monolayers to hyposmotic Ringer solution. To evaluate the possible role of the cytoskeleton in the regulation of Na+ channels in response to a change in osmolality we used four different experimental approaches. In the control group, which were not exposed to any cytoskeleton-influencing drugs, there was a 1.5-fold increase in Isc and in the number of open Na+ channels after osmotic stimulation. For the second group cytochalasin D (0.1 microg/ml) was present on the serosal side during the experiments. Neither Isc nor the number of open Na+ channels increased after osmotic stimulation. In the third group colchicine (0.2 mM) or nocodazole (20 microM) was present on the serosal side, which resulted in 1.8-fold and 1.5-fold increases in Isc as well as 3-fold and 2-fold increases in the number of Na+ channels, respectively. In the fourth experimental group erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (EHNA, 0.5 mM), a dynein inhibitor, was present on the serosal side. In this group Isc decreased to about 0.4 microA/cm2, and subsequent application of amiloride abolished Isc completely. Under hyposmolar conditions EHNA abolished entirely the sensitivity of Isc to the osmotic challenge. Because of the EHNA-induced down-regulation of Isc, the density of apical Na+ channels in this experimental group could not be determined. These results show that the cytoskeleton is dominantly involved in osmotic channel regulation at the apical membrane, and that actin filaments, microtubules and molecular motors are involved in the recruitment of additional Na+ channels.

Influence of fish meal quality and feed pellet on growth, feed efficiency and muscle composition in gilthead seabream ( sparus aurata)

The effect of fish meal quality on growth, feed efficiency, protein digestibility and fillet composition was studied in a growth experiment with gilthead seabream. The experiment included a comparison of pelleted feed with extruded feed. The fish were fed three extruded diets which varied in the quality of the fish meal used in the respective feeds: low quality fish meal, good quality fish meal, and a mixture (50%) of the two fish meal qualities. Fish meal quality was judged by protein digestibility as measured with mink as the test animal and by the content of biogenic amines. A fourth experimental group was fed a pelleted feed with the same mixed fish meal blend. The experiment started with 70 g seabream and lasted for 3 months until the fish reached about 160 g. All groups showed good growth and feed efficiency during the experimental period, with daily specific growth rates (SGR) of 0.90–1.00% and feed efficiencies (FE; fish weight gain per feed offered) from 0.58 to 0.66. The fish fed the feed with low quality fish meal showed significantly poorer feed efficiency than the other groups, and there was a significant correlation between feed efficiency obtained in seabream and the protein digestibility as measured with mink. No difference was observed among groups fed the extruded diets, for growth, fillet composition or amount of liver or viscera, although a significant correlation was obtained between SGR and protein digestibility in mink. Fillet lipid content was higher in fish fed the extruded feed, compared with those fed the corresponding pelleted diet. Protein digestibilty in seabream showed no difference among the extruded feeds, while the pelleted feed was significantly lower.

Intracellular Ca2+, intercellular electrical coupling, and mechanical activity in ischemic rabbit papillary muscle. Effects of preconditioning and metabolic blockade.

During myocardial ischemia, electrical uncoupling and contracture herald irreversible damage. In the present study, we tested the hypothesis that an increase of intracellular Ca2+ is an important factor initiating these events. Therefore, we simultaneously determined tissue resistance, mechanical activity, pH(0), and intracellular Ca2+ (with the fluorescent indicator indo 1, Molecular Probes, Inc) in arterially perfused rabbit papillary muscles. Sustained ischemia was induced in three experimental groups: (1) control, (2) preparations preconditioned with two 5-minute periods of ischemia followed by reperfusion, and (3) preparations pretreated with 1 mmol/L iodoacetate to block anaerobic metabolism and minimize acidification during ischemia. In a fourth experimental group, intracellular Ca2+ was increased under nonischemic conditions by perfusing with 0.1 mmol/L ionomycin and 0.1 mumol/L gramicidin. Ca2+ transients and contractions rapidly disappeared after the induction of ischemia. In the control group, diastolic Ca2+ began to rise after 12.6 +/- 1.3 minutes of ischemia; uncoupling, after 14.5 +/- 1.2 minutes of ischemia; and contracture, after 12.6 +/- 1.5 minutes of ischemia (mean +/- SEM). Preconditioning significantly postponed Ca2+ rise, uncoupling, and contracture (21.5 +/- 4.0, 24.0 +/- 4.1, and 23.0 +/- 5.3 minutes of ischemia, respectively). Pretreatment with iodoacetate significantly advanced these events (1.9 +/- 0.7, 3.6 +/- 0.9, and 1.9 +/- 0.2 minutes of ischemia, respectively). In all groups, the onset of uncoupling always followed the start of Ca2+ rise, whereas the start of contracture was not different from the rise in Ca2+. Perfusion with ionomycin and gramicidin permitted estimation of a threshold [Ca2+] for electrical uncoupling of 685 +/- 85 nmol/L. In conclusion, the rise in intracellular Ca2+ is the main trigger for cellular uncoupling during ischemia. Contracture is closely associated with the increase of intracellular Ca2+ during ischemia.

A study of the apical microleakage of a gallium alloy as a retrograde filling material

The feasibility of utilizing mercury-free Gallium alloy GF for retrograde filling was investigated by comparing apical microleakage in 184 extracted human teeth. The teeth were divided into four experimental and two control groups. Three experimental groups were apical cavity retrofillings with the Gallium alloy GF, a mercury-containing amalgam, and a glass ionomer. The fourth experimental group was filled with gutta-percha and heat-burnished after apicoectomy. After 24 h, 1 wk, 4 wk, and 12 wk immersion in dye solution, the roots were vertically sectioned, and the deepest point of dye penetration was recorded. The glass ionomer showed the least leakage, followed by the amalgam group and the gallium group (no significant difference). The gutta-percha heat-burnished group displayed the greatest leakage. Gallium alloy GF was shown to have an equivalent sealing potential to dental amalgam for a retrograde filling material.

Decreased duration of emergency department treatment of chronic obstructive pulmonary disease exacerbations with the addition of ipratropium bromide to β-agonist therapy

To determine the benefit of the addition of ipratropium bromide to beta-agonist therapy of acute exacerbations of chronic obstructive pulmonary disease. The trial was randomized and double blinded. The study was conducted in the emergency department of Parkland Memorial Hospital, a busy, inner-city, county hospital. Patients were treated in the medicine emergency department with either the standard regimen of nebulized isoetharine, 0.5 mL of a 1% solution (5.0 mg) diluted to 2.0 mL with normal saline every hour (control group) or with the same regimen plus ipratropium bromide, 54 micrograms (three puffs) after the first isoetharine treatment and 36 micrograms (two puffs) after the second and fourth (experimental group). A placebo metered-dose inhaler used in the same manner as the ipratropium blinded the study to both the patients and medical personnel. The group treated with the addition of ipratropium (30) was discharged from the ED an average of 91 minutes (P less than .05) sooner than the control group (25) and required on the average one less isoetharine treatment (P less than .05). The pulmonary functions tested, forced expiratory volume in the first second, and the forced vital capacity were the same in the two groups initially and on discharge, as identical discharge criteria were used in each group. The addition of ipratropium to standard beta-agonist treatment of chronic obstructive pulmonary disease exacerbations shortens the duration of treatment required in the ED.

Organization of efferent projections of the subthalamic nucleus in the squirrel monkey as revealed by retrograde labeling methods

The cellular origin and degree of collateralization of the subthalamostriatal, subthamonigral and subthalamopallidal projections in the squirrel monkey ( Saimiri sciureus) were studied using lectin-conjugated horseradish peroxidase (WGA-HRP), Nuclear yellow (NY) and Fast blue (FB) as retrograde tracers. In a first experimental group, WGA-HRP was injected in the left putamen and the right caudate nucleus. Followingl these injections numerous retrogradely labeled neurons occured in the dorsolateral two-thirds of the subthalamic nucleus on the putamen-injected side, whereas a smaller numebr of positive cells were found in the ventromedial third of the same nucleus on the caudate-injected side. In a second experimental group NY was injected in the putamen whereas FB was delivered in the substantia nigra on the same side of the brain. After putaminonigral injections subthalamic cells containing the tracer injected in the putamen (about 75–80% of all retrogradely labeled neurons) occured in the dorsolateral two-thirds of the nucleus, whereas those containing the tracer injected in the substantia nigra (about 20–25% of all positive subthalamic cells) were confined to the ventromedial third of the structure. Approximately 5–10% of all subthalamic positive neurons were double-labeled following putaminonigral injections. In a third experimental group, NY was injected in the caudate nucleus and FB in the substantia nigra on the same side. After such injections cells retrogradely labeled with NY or FB were present in about equal number and appeared closely intermingled in the ventromedial third of the subthalamic nucleus. Less than 10% of all positive subthalamic neurons were double-labeled following caudatonigral injections. In a fourth experimental group, NY was delivered in the globus pallidus and FB in the substantia nigra on the same side. In these animal cells containing the tracer delivered in the pallidum were about 4 times more numerous than those labeled with the tracer injected in the substantia nigra, and approximately 10–20% of all positive subthalamic neurons were double-labeled following pallidonigral injections. Most of these double-labeled cells occured in the zone where the two populations of single-labeled cells overlapped. Finally, in a last experimental group, NY was injected in the pedunculopontine nucleus on one side and in the substantia nigra on the other. Following these injections cells containing the tracer delivered in the pedunculopontine nucleus were found to be 5–6 times less numerous than those labeled after substantia nigra injections. These findings suggest that in regard to its efferent projections, the primate subthalamic nucleus is organized according to a complex pattern consisting of: (1) a large (80%) dorsolateral ‘sensorimotor’ zone where most neurons project to the lenticular nucleus and terminate in the globus pallidus and/or the putamen. (2) a small (20%) ventromedial ‘associative’ zone neurons give to either ascending projections to caudate nucleus or descending projections to the substantia nigra, and (3) a smaller (10%) overlapping zone whose neurons send axon collaterals to both the lenticular nucleus and the substantia nigra.

Corticosterone Rhythmicity in the Rat: Interactive Effects of Dietary Restriction and Schedule of Feeding

Effects of changes in diet composition and schedule of restricted feeding on the circadian rhythm of plasma corticosterone in the adult rat were examined. Three restricted feeding schedules were used: diet presented 3 hours after lights-on, 1 hour before lights-off or continuously by automated feeder. Animals were paired to controls (consuming 24% protein diet ad libitum) and provided 80% of control food intake/g body weight/day. A fourth experimental group was offered 2.4% protein diet ad libitum. Plasma corticoids were measured every 4 hours from 0800 to 0400 hours. Corticoid levels 20 min after food presentation were also determined. Both control and protein-restricted animals displayed a typical circadian rhythm, with corticoids lowest at 0800 hours and a peak at 2000 hours. Animals fed at 0900 hours exhibited a 12-hour shift in peak corticoids, while the circadian rhythm was totally abolished in animals fed by automated feeder. Offering the restricted ration at 1700 hours, however, resulted in a diurnal rhythm similar to that of controls. While pre and post-feeding corticoids were similar in control and protein-restricted animals, a post-feeding suppression was found in all three food-restricted groups. Clearly, the relationship between feeding schedule and diurnal rhythm must not be ignored in nutritional paradigms.calorie restriction circadian rhythm corticosterone