Effects of supplemental dietary tannins on the performance of white-tailed deer (Odocoileus virginianus).

Tannins are natural and nutritionally significant components of the diets of browsing ungulates. In trials on supplemented pastures and in drylots, we estimated dry matter intake (DMI), weight gain, and urea N, potassium, cortisol and creatinine in urine of captive white-tailed deer fed pelleted diets that differed only in the respective quebracho tannin (QT) content. The low control, medium and high QT rations were 3.6, 63 and 152 g/kg DM respectively. There was no tannin-free pellet option. Trials were divided into winter pasture, restricted choice and spring growth. In winter pasture trial on pasture using QT, deer reduced QT intake relative to that expected under random foraging. This aversion was also apparent during the spring growth trial. While DMI in the winter pasture trial remained similar among treatments (p > 0.05), averaging 130 g/kg(0.75)/day, deer gained more weight (p < 0.05) when given a choice that included the high QT ration. During subsequent spring growth, DMI and weight gains generally exceeded those of the winter period. Unlike the winter pasture trial, weight gains in spring growth trial were higher (p < 0.05) in the low-control QT treatment. In the restricted choice trial, weight gain was again higher (p < 0.05) for deer fed a low-control QT diet. The urea N/creatinine ratio of deer fed the low-control QT diet (0.0357) was over three times that of deer fed the high QT diet (0.0107). Neither potassium/creatinine nor cortisol/creatinine ratios were affected by diet (p > 0.05). Collectively, these results suggest that although deer do not avoid tannins, and even ingested up to 5% under the choice options in these trials, the effect of tannins on deer performance may vary by season as well as by foraging opportunities.

Performance and dietary preferences of white-tailed deer grazing chicory, birdsfoot trefoil or alfalfa in north central Alberta.

Little information exists on the performance of deer on alternative forage species in northern temperate environments during summer and fall, the period of inherent maximum growth in deer. In performance and choice experiments, we compared live weight gain (g/kg(0.75)/day), absolute [kg/ha dry matter (DM)] and relative (% DM) herbage utilization, relative preference index (RPI) as well as plant community visitation of white-tailed deer grazing alfalfa (Medicago sativa), birdsfoot trefoil (Lotus corniculatus) or chicory (Cichorium intybus) in north central Alberta, Canada. Herbage phytomass and quality was also measured on the grazed pastures. Alfalfa had higher dry matter yields and crude protein concentrations than chicory and trefoil. Chicory had lower neutral detergent fiber concentrations than the other forages. Tannin concentrations were greatest in birds foot trefoil (nearly 55 g/kg DM), well above those in the other forages (<5 g/kg DM). Live weight gain was similar among deer feeding within the paddocks seeded to birds foot trefoil and chicory, and more than two times higher (p < 0.05) than deer feeding in paddocks seeded to alfalfa. Deer spent more grazing time (about 40%) on chicory pastures than on alfalfa and birds foot trefoil pastures. RPI values were greatest for birds foot trefoil at 2.11, intermediate for chicory at 1.40, and lowest for alfalfa at <0.60. Absolute herbage utilization remained similar (p > 0.05) among the three forage species. In contrast, relative herbage utilization was greater from birds foot trefoil (52% DM) than chicory (40% DM) or alfalfa (25% DM). These results suggest that the use of alfalfa with other alternative forages may prove beneficial to deer production, rather than using alfalfa pasture alone.

Visualisation of needle position using ultrasonography.

Anaesthetists and intensivists spend a considerable proportion of their working time inserting needles and catheters into patients. In order to access deeper structures like central veins and nerves, they have traditionally relied on surface markings to guide the needle into the correct position. However, patients may present challenges due to anatomical abnormalities and size. Irrespective of the skill of the operator, there is the ever-present risk of needle misplacement with the potential of damage to structures like arteries, nerve bundles and pleura. Repeated attempts, even if ultimately successful, cause patient suffering and probably increase the risk of infection and other long term complications. Portable and affordable, high-resolution ultrasound scanners, has accelerated the interest in the use of ultrasound guidance for interventional procedures. Ultrasound guidance offers several advantages including a greater likelihood of success, fewer complications and less time spent on the procedure. Even if the target structure is identified correctly there is still the challenge to place the needle or other devices in the optimum site. The smaller and deeper the target, the greater the challenge and potential usefulness of ultrasound guidance. As a result of limited training in the use of ultrasound we believe that many clinicians fail to use it to its full potential. A lack of understanding, with regard to imaging the location of the needle tip remains a major obstacle. Needle visualisation and related topics form the basis for this review.

Free cortisol index is better than serum total cortisol in determining hypothalamic-pituitary-adrenal status in patients undergoing surgery.

Serum total cortisol has traditionally been used for the interpretation of tests of the hypothalamic-pituitary-adrenal axis. Approximately 80% of total cortisol is bound to cortisol-binding globulin (CBG), and variation in CBG significantly affects serum total cortisol levels. Reliable assessment of hypothalamic-pituitary-adrenal axis reserve is difficult in severely ill patients, because CBG falls substantially during the acute phase response. The free cortisol index (FCI), defined as the ratio of total cortisol/CBG, correlates well with serum free cortisol. We evaluated the FCI in the context of severe stress and the acute phase response by measuring total cortisol and CBG pre- and postoperatively in 31 patients undergoing major elective surgery. Serum total cortisol increased by 55% from 453 +/- 35.2 (mean +/- SEM) nmol/liter (range, 88-882) to 700 +/- 47.2 (range, 294-1631) nmol/liter. Serum CBG decreased by 30% from 45 +/- 1.7 (range, 26.6-64.1) to 31.4 +/- 1.62 (range, 16.1-51.9) mg/liter, but FCI increased by 130% from 10 +/- 0.8 (range, 2-18) to 23 +/- 1.7 (range, 13-58) nmol/mg. In seven patients (23%), postoperative serum total cortisol was less than 500 nmol/liter, but their postoperative CBG levels were significantly lower than levels in the rest of the group (P < 0.01). However, there was no difference in the FCI between this subgroup and the rest of the group. This study demonstrates the importance of CBG measurement and the calculation of FCI for the interpretation of serum total cortisol in situations where CBG changes significantly.

ASP1 (BACE2) cleaves the amyloid precursor protein at the beta-secretase site.

Sequential proteolytic processing of the Amyloid Precursor Protein (APP) by beta- and gamma-secretases generates the 4-kDa amyloid (A beta) peptide, a key component of the amyloid plaques seen in Alzheimer's disease (AD). We and others have recently reported the identification and characterisation of an aspartic proteinase, Asp2 (BACE), as beta-secretase. Here we describe the characterization of a second highly related aspartic proteinase, Asp1 as a second beta-secretase candidate. Asp1 is expressed in brain as detected at the mRNA level and at the protein level. Transient expression of Asp1 in APP-expressing cells results in an increase in the level of beta-secretase-derived soluble APP and the corresponding carboxy-terminal fragment. Paradoxically there is a decrease in the level of soluble A beta secreted from the cells. Asp1 colocalizes with APP in the Golgi/endoplasmic reticulum compartments of cultured cells. Asp1, when expressed as an Fc fusion protein (Asp1-Fc), has the N-terminal sequence ALEP..., indicating that it has lost the prodomain. Asp1-Fc exhibits beta-secretase activity by cleaving both wild-type and Swedish variant (KM/NL) APP peptides at the beta-secretase site.

Fractalkine cleavage from neuronal membranes represents an acute event in the inflammatory response to excitotoxic brain damage.

Fractalkine is a recently identified chemokine that exhibits cell adhesion and chemoattractive properties. It represents a unique member of the chemokine superfamily because it is located predominantly in the brain in which it is expressed constitutively on specific subsets of neurons. To elucidate the possible role of neuronally expressed fractalkine in the inflammatory response to neuronal injury, we have analyzed the regulation of fractalkine mRNA expression and protein cleavage under conditions of neurotoxicity. We observed that mRNA encoding fractalkine is unaffected by experimental ischemic stroke (permanent middle cerebral artery occlusion) in the rat. Similarly, in vitro, levels of fractalkine mRNA were unaffected by ensuing excitotoxicity. However, when analyzed at the protein level, we found that fractalkine is rapidly cleaved from cultured neurons in response to an excitotoxic stimulus. More specifically, fractalkine cleavage preceded actual neuronal death by 2-3 hr, and, when evaluated functionally, fractalkine represented the principal chemokine released from the neurons into the culture medium upon an excitotoxic stimulus to promote chemotaxis of primary microglial and monocytic cells. We further demonstrate that cleavage of neuron-derived, chemoattractive fractalkine can be prevented by inhibition of matrix metalloproteases. These data strongly suggest that dynamic proteolytic cleavage of fractalkine from neuronal membranes in response to a neurotoxic insult, and subsequent chemoattraction of reactive immune cells, may represent an early event in the inflammatory response to neuronal injury.

The role of fractalkine in the recruitment of monocytes to the endothelium.

Recombinant fractalkine possesses both chemoattractive and adhesive properties in vitro. Previous studies have demonstrated an upregulation of this molecule on the membranes of activated human endothelial cells and hypothesised that fractalkine plays a role in the recruitment and adherence of monocytes to the activated endothelium. Here we present data analysing both the adhesive and chemoattractive properties of this chemokine expressed by activated human umbilical vein endothelial cells. We demonstrate that both recombinant fractalkine and endogenously produced fractalkine function as adhesion molecules, tethering monocytes to the endothelium. However, our data demonstrate that although recombinant fractalkine has the potential to function as a potent monocyte chemoattractant, the endogenous fractalkine cleaved from activated human umbilical vein endothelial cells is not responsible for the observed chemotaxis in this model. Instead, we show that monocyte chemoattractant protein-1 (MCP-1), secreted from the activated human umbilical vein endothelial cells, is responsible for the chemotaxis of these monocytes.

Structural and functional analysis of the rat metallothionein III genomic locus.

Metallothionein III (MT III) has been reported to suppress neuronal growth in a rat in vitro model system. The protein and its specific mRNA are detected predominantly in the brain, differentiating MT III from the well-characterised archetypal metallothioneins. Isolation, sequencing and functional analysis of the rat MT III genomic locus indicated that, although the organisation of the gene was conserved between MT III and the more conventional metallothioneins, the 5' flanking region of the MT III gene was distinct. Within this region, a number of putative regulatory elements were identified, including the metal regulatory elements (MREs) characteristic of metallothionein promoters. However, despite their conservation in sequence with active elements, the MREs of MT III were unresponsive to zinc. A 'silencing element' was revealed within a 250 bp section of the MT III promoter which suppressed gene expression in two brain cell lines. The operation of this silencing region in conjunction with the inactive MREs may explain the distinct expression profile observed for MT III within the central nervous system and during neuronal development.

Improving the prescribing of antibiotics for urinary tract infection.

BACKGROUND: In recent years there have been changes in the recommended antibiotic treatment for urinary tract infections (UTIs). In particular, the use of amoxycillin or co-trimoxazole is now discouraged, with amoxycillin-potassium clavulanate, cephalexin and trimethoprim becoming first-line agents for uncomplicated lower UTIs. AIM: To examine whether academic detailing, performed by a pharmacist, could modify prescribing practices for antibiotics used in the treatment of UTI in the community setting. METHODS: The intervention was conducted in Southern Tasmania, using the remainder of the State as a control area. The target group of general practitioners was sent educational material designed to assist in the appropriate prescribing of antibiotics in the treatment of UTI. A pharmacist then visited each general practitioner and discussed the rational use of antibiotics for UTIs directly with him/her. Outcomes were measured using evaluation feedback from the general practitioners and pharmacoepidemiological data, which were not linked to diagnosis. The key variable examined was the total defined daily doses (DDDs) dispensed for the recommended first-line agents (amoxycillin-potassium clavulanate, cephalexin and trimethoprim) compared with amoxycillin (3 g single-dose form) and co-trimoxazole. RESULTS: The educational programme was very well received by the general practitioners. Changes in the prescribing of antibiotics commonly used for UTIs were evident in both study regions over the course of the study, but the improvements were significantly greater in the intervention area. CONCLUSION: Educational programmes utilizing academic detailing by pharmacists can modify prescribing practices within the community setting.

Effect of low dissolved oxygen on aquatic life stages of the caddisfly Clistoronia magnifica (Limnephilidae).

Embryos, larval stages (instars I-V), pupal stages, and pharate adults of the caddisfly Clistoronia magnifica (Limnephilidae) were exposed to a range of dissolved oxygen (DO) concentrations (0.9-8.3 mg/L) for 4-88 days in the laboratory. Some embryos suspended growth at low DO, resuming growth and hatch when DO was increased. Embryos and larvae all had 96-h EC50 values (50% mortality at 96 h) of about 2.0 mg/L DO. The statistical Effect and No-Effect Thresholds for larvae exposed through two molts from instars I-III were 1.6 and 2.4 mg/L, respectively. At DO concentrations below 4.6 mg/L, egg hatch, larval development, molting success, time of molting, pupation, and adult emergence were delayed.

Ovine tracheal muscle contraction in vitro: inhibition by calcium channel blockers gallopamil and verapamil.

We compared the inhibitory effects of calcium channel blockers, gallopamil and verapamil on acetylcholine (Ach)-induced contractions of ovine tracheal muscle in vitro. Adult sheep were sacrificed and tracheal strips were obtained by cutting the single tracheal rings from the mid-trachea. Tracheal strips were suspended in Krebs-Henseleit solution and isometric tension measured upon stimulation with cumulative doses of Ach (10(-7) to 10(-4) M) without and after pretreatment with gallopamil (10(-7) to 10(-6) M) or verapamil (10(-6) to 10(-5) M). In untreated tissues, the mean concentration of Ach required to produce 50% of maximal response (EC50) was 4.3 x 10(-6) M Ach. Both gallopamil and verapamil inhibited the Ach-induced contractions of ovine tracheal smooth muscle, by shifting the dose-response curves to Ach to the right. EC50 Ach for gallopamil (10(-6) M) and verapamil (10(-6) M) was 2.6 x 10(-5) and 5.2 x 10(-6) M, respectively. Dose ratio defined as postantagonist EC50 Ach/control EC50 Ach, was 7.7 for gallopamil and 2.0 for verapamil. Thus, the inhibitory effect of gallopamil was approximately 4-fold more potent than that of verapamil. Gallopamil was 17-fold more potent than verapamil in relaxing precontracted tracheal strips. The dose of calcium antagonists required to produce 25% relaxation (EC25) of tracheal strips precontracted with 10(-4) Ach was 3.7 x 10(-5) M for verapamil and 2.2 x 10(-6) M for gallopamil. These results indicate that gallopamil is effective against Ach-induced contractions of ovine trachealis muscles, and is more potent than verapamil.

Cellular LTB4 production differs in allergic sheep with and without late airway responses.

We tested the hypothesis that allergic sheep that develop both early and late airway responses to inhaled Ascaris suum antigen (late responders) have an increased capacity to generate leukotrienes (LTs) compared with allergic sheep that show only early responses to inhaled antigen (acute responders). To test this hypothesis, we measured LTB4 production, in vitro, by granulocytes isolated from peripheral blood and by macrophages isolated from bronchoalveolar lavage (BAL) from both groups of sheep greater than or equal to 2 wk after the animal's last antigen challenge; LTB4 production by granulocytes isolated from BAL from both groups of sheep 6 and 48 h after local airway challenge with A. suum antigen was also measured. LTB4 production was induced by incubating cells (i.e., either granulocytes or macrophages) with calcium ionophore (A23187, 2 microM) and arachidonic acid (30 microM). LTB4 production was quantitated by high-performance liquid chromatography and verified by radioimmunoassay (RIA). On stimulation peripheral blood granulocytes from late responders (n = 7) produced (means +/- SD/10(6) cells) 13.3 +/- 5.2 ng LTB4 compared with 5.3 +/- 1.5 ng LTB4 (P less than 0.05) for acute responders (n = 7). This increased LTB4 production did not result from variations in granulocyte differential or cyclooxygenase activity (as indicated by RIA measurements of prostaglandin E2 production).(ABSTRACT TRUNCATED AT 250 WORDS)

Removal of bronchial secretions by two-phase gas-liquid transport.

Net outward wave motion of secretions from airways by two-phase gas-liquid transport is favored by higher airflow during expiration than inspiration. This can be accomplished by IRV in which the controlled mode of mechanical ventilation is adjusted such that the inspiratory cycle is prolonged and the expiratory phase is shortened. Studies were done on six anesthetized, nasally intubated sheep. Simulated mucus was instilled into the bronchi at 15-min intervals during I-E ratio of 1:2.7, 1.9:1, and 3:1. The IRV modes of 1.9:1 and 3:1 promoted transport of simulated mucus outward. Neither systemic blood pressure nor cardiac output were altered by IRV. Thus, IRV might be useful in the management of excessive bronchial secretions in mechanically ventilated patients. However, clinical trials of IRV should take into account its potential for producing adverse hemodynamic effects and barotrauma in patients with compromised cardiac function due to auto-PEEP attendant with its usage.

Effect of chest wall oscillation on mucus clearance: comparison of two vibrators.

This study was designed to investigate the effect of an experimental low-energy chest wall oscillator and of a commercial chest percussor on central airway mucociliary clearance. Five normal dogs were anesthetized, intubated, and placed supine in a trough to which the oscillator or percussor was mounted. Tracheal mucus velocity (TMV) was measured by radiopaque particle or charcoal spot movement. The commercial percussor (a fixed sinusoidal device) used at its minimum frequency of 40 Hz, produced a mean (+/- SE) maximum expiratory flow rate of 0.25 +/- 0.04 L/sec at the airway opening, and had no measurable effect on TMV. The experimental oscillator, when operated at a level sufficient to generate flows of 2-3 L/sec, and with an unbiased 13-Hz sine wave (estimated energy, 150 W), increased mean TMV to 204 +/- 13% of control (P less than 0.003); the percent increase was independent of baseline TMV. We conclude that moderate oscillatory power applied to the chest wall can enhance mucus clearance in central airways, but that currently available commercial percussors may not meet the mechanical requirements for this effect.

Cardiopulmonary effects of direct venous CO2 insufflation in ewes. A model for CO2 hysteroscopy.

In order to mimic the delivery of CO2 into the pelvic circulation during sustained hysteroscopic surgery, direct insufflation of CO2 was done into the femoral vein of six anesthetized ewes. Following the establishment of baseline values in each animal, cardiac output, pulmonary arterial pressure (Ppa), pulmonary arterial wedge pressure (Ppw), arterial pH and gas levels, and ECG changes were recorded at ten-minute intervals. CO2 was delivered into the femoral circulation for 30 minutes. Following the experiment, measurements were repeated during a 20-minute recovery period. Ppa and Ppw increased significantly during the experiment but returned to baseline values after 70 minutes. Cardiac output, which increased significantly, remained high after the same period and was paralleled by cardiac rate. There was no significant change in systemic blood pressure or arterial oxygenation. Only at the highest flow rate were there observable changes in PCO2, accompanied by mild acidosis. One of the six animals displayed premature ventricular contractions at the inception of the highest flow rate. Since the delivery rate of CO2 per kilogram of body weight clearly exceeded that generally used in human hysteroscopic surgery (35-100 mL/min), these experimental results suggest that CO2, when employed as a distension modality in hysteroscopic surgery, displays a wide margin of safety.

The effect of nedocromil sodium and cromolyn sodium on antigen-induced responses in allergic sheep in vivo and in vitro.

We studied the effects of nedocromil sodium and cromolyn sodium on early and late bronchial responses to inhaled Ascaris suum antigen in allergic sheep in vivo, and the antigen-induced contractile responses of sheep tracheal smooth muscle in vitro. For the in vivo studies the sheep were pretreated with aerosols of placebo (buffered saline solution), 20 mg of nedocromil sodium, or 20 mg of cromolyn sodium (both dissolved in 3 ml of buffered saline solution) and then challenged with aerosol antigen. Specific pulmonary resistance (SRL) was measured before and after challenge to document the responses of the airways. In the trial with placebo, challenge with antigen resulted in significant early and late increases in SRL. Treatment with nedocromil sodium significantly reduced the early response to antigen and blocked the late response. Cromolyn sodium gave the same results; there were no statistical differences between the responses of the airways for the two dogs. In vitro nedocromil sodium at doses of 10(-6)M and 10(-5)M inhibited significantly the contractile responses of sheep tracheal smooth muscle to A suum. Cromolyn sodium only showed efficacy at 10(-5)M. These results suggest that nedocromil sodium may be potentially useful in the treatment of reversible allergic disease of the airways.

Laser smoke effect on the bronchial system.

The photoresection of endobronchial tumors produces smoke which is partly inhaled by the patient as well as the surgical staff. In an animal study we investigated whether a single exposure or repetitive exposures to smoke might have harmful side effects on the airways. Eleven sheep were exposed to smoke produced by laser-vaporizing (6,500 J) sections of bronchial tissue (1 cm3) in a Plexiglas chamber. The smoke analysis revealed 0.92 mg/liter particles with a mean particle size of 0.54 micron. Carbon monoxide content was estimated as 0.04%. We measured the effects of one or three separate ten-minute exposures on airway resistance, gas exchange, and mucociliary clearance rate in the trachea. We found that the smoke inhalation resulted in a decrease of arterial PO2 with relatively little change in airway mechanics. Tracheal mucus velocity, a marker of lung mucociliary clearance, was significantly depressed in a dose-dependent manner with increasing smoke exposures. Results of bronchoalveolar lavages performed before and one day after the exposure showed that the smoke inhalation induced a severe inflammation with dramatic increases of inflammatory cells. The total number of cells per milliliter lavage return increased from 3.2 million to 25 million; percent neutrophils increased from 2.3 to 45.6% and percent macrophages decreased from 86 to 41%. These findings indicate that the side effects of smoke inhalation during intrabronchial laser surgery should not be neglected. The impairment of the defense mechanism of the lung combined with the inflammation as well as hypoxia might be fatal in compromised patients. Effective smoke removal devices should be developed to protect the patient as well as the surgeon.

The effect of an orally active leukotriene D4/E4 antagonist, LY171883, on antigen-induced airway responses in allergic sheep.

Leukotriene (LT) D4 is a putative mediator of allergic asthma: inhaled LTD4 produces early and late increases in specific lung resistance (SRL) and slows tracheal mucus velocity (TMV) similar to inhaled antigen. In this study we examined the effects of an orally active LTD4/LTE4 antagonist, LY171883 [1-less than 2-Hydroxy-3-propyl-4-less than 4-(1H-Tetrazol-5-yl) Butoxy greater than Phenyl greater than Ethanone], on early and late changes in SRL and TMV following airway challenge with Ascaris suum antigen in conscious allergic sheep. SRL and TMV were measured before and up to 8 h and 24 h after antigen challenge after either LY171883 (30 mg/kg, p.o. 2 h before challenge) or placebo pretreatment. After placebo pretreatment antigen challenge resulted in significant early (483% over baseline) and late (221% over baseline) increases in SRL (n = 9). LY171883 pretreatment, however, significantly reduced the early increase in SRL (163% over baseline) and blocked the late response. LY171883 did not prevent the antigen-induced fall in TMV from 5-8 h post challenge (n = 6), but TMV recovered more rapidly in the drug trial returning to baseline values by 24 h. These results suggest that the generation of LTD4, and its metabolite LTE4, during airway anaphylaxis contributes to the early increase in SRL and is important for eliciting the late increase in SRL as well as contributing to the fall in TMV.