Organization of the optic chiasm in the hatched chick.

In the hatched chick the fibers of the two optic nerves segregate into clearly defined bundles when they cross to the other side. These bundles run in horizontally oriented tiers. The tiers are demarcated by blood vessels and pial tissue. The organization of these tiers was investigated qualitatively and quantitatively using light and electron microscopy as well as tracer techniques. The fibers within the tiers cross to the contralateral optic tract without leaving their respective tier. The mean total number of tiers is 34 with a great individual variation. A preference in the superposition of one side over the other could not be observed. Comparing these data with our earlier study (Rager et al.: Anat. Embryol., 179:135-148, 1988) it can be concluded that neither the segregation of fibers into discrete bundles nor the variability in the number of alternating tiers seem to disturb the topography of fibers as it is achieved in the optic nerve. The pattern of vascularization correlates with the order of crossing axon bundles and contributes to the demarcation of the tiers. The chiasm is vascularized by the Aa. preopticae and the A. infundibularis.


Accuracy and precision of computerized models of the anterior cranial base in young mice.

The quantitative analysis of craniofacial growth in experimental animals relies on computerized reconstructions in order to measure changes in form. The purpose of this study was to determine the validity of computerized three-dimensional models of the anterior cranial base in mice. Ten 1-day-old non-littermates were collected and the anterior cranial base was dissected free from surrounding connective tissues. Eleven measurements were recorded from these cartilages. Twenty developmentally equivalent mice were collected and fixed with either glutaraldehyde or formalin and the anterior cranial base from each specimen was subjected to computerized reconstruction. The corresponding 11 measurements were recorded from these models. Results showed that the measurements recorded from the computerized models were not significantly different from those recorded directly from the actual anterior cranial bases. Therefore, the reconstructions were considered accurate. An analysis of the coefficients of error revealed that measurements derived from the computerized models were significantly more precise than those recorded directly from the actual tissue. The computerized three-dimensional reconstruction method provides accurate and precise models of the anterior cranial base in young mice.


[The use of cryopreserved swine blood for in vitro feeding of the soft tick Ornithodoros moubata].

Porcine stored blood has been used in the in vitro feeding of the soft tick Ornithodoros moubata through a Parafilm membrane. The efficiency of the feeding decreased after storage of 5 weeks by -20 degrees C. Through addition of 10(-3) M ATP/l the feeding results with frozen blood for duration of storage up to 200 days well agreed with the results of feeding tests with fresh porcine blood: The nymphs (N)1-3 fed the 4-5fold, the N4-6 the 2.7fold and the adults (A) the 1.9fold of their body weight. The feeding rate was 98.6% (N1-3); 92.7% (N4-5) and 81.9% (A). The mortality rate was under 10%. Females laid 130 eggs after blood meal.


[The occurrence of Echinococcus granulosis and E. multilocularis in Thuringia].

The occurrence of E. granulosus and E. multilocularis in the region of Thuringia is reported. Parasitological investigations showed 1421 E. granulosus metacestodes, 91.7% of them in lungs and 1.3% in livers of cattle, 6% in lungs and 1% in livers of pigs; that means an infestation rate at slaughter of 0.1%-0.3% in cattle and 0.001-0.004% or less in pigs resp. 90.1% of the hydatid cysts proved to be fertile even in a size of 1.5 cm diameter. Adult E. granulosus was found post mortem in 2 of 324 dogs. In the period from 1985 to 1988, only 11 dogs were infested with E. granulosus as found at autopsy all over the GDR. Out of 23,325 faecal samples 270 samples (1.2%) were positive for eggs of Taenia spp. The animals with egg-shedding were treated as infected with Echinococcus. In experimental infections of 12 Beagles the prepatent period ranged from the minimum of 34 days up to the maximum of 40 days. The detected E. granulosus strain could be identified as a dog-cattle strain. The microscopical examination of the intestine of 805 red foxes (Vulpes vulpes) revealed the presence of E. multilocularis in 12.7% of the animals. Thuringia is one of the endemic distribution area of E. multilocularis in Central Europe. In the west of Thuringia 25% of the foxes were found with E. multilocularis, in the remaining area (lowlands) only 3.3%. In some of mountainous areas, 40% of the foxes were infected with E. multilocularis. 2 cats of 58 from this region were infected with E. multilocularis.


Behavioural aspects and their possible uses in the control of dracontiasis (guinea-worm) in Igwun river basin area of Imo State, Nigeria.

Individuals suffering from dracontiasis from Igwun river basin area of Imo State Nigeria were randomly chosen, examined and interviewed between December 1988 and March 1989 with a view to ascertain some of the behavioural aspects that could be of help in the control/eradication of this disease as well as to ascertain whether local medication was of any chemotherapeutic significance. Of 100 guinea-worm patients males were significantly higher (P < 0.05) in number (63.0%). In the age-related distribution, children less than 10 years old and the members of the villages more than 60 years old accounted for only 5.0% of the patients whilst those in their productive years (10-60 year old) accounted for 95.0%. Only 10.5% of the respondents associated guinea-worm infection with the drinking of "polluted" water while most believed it was a familial trait (36.8%) or implicated their enemies (35.1%). As a result medications against the disease were mainly directed towards consulting the oracle and herbalists, and appeasing the gods. Most (98.0%) of them kept their dressing dry by refraining from immersing them in water. The concomitant behavioural aspects of these results are discussed in relation to their uses in the control/eradication of guinea-worm.


Human helminthosis in a rural community of Plateau State, Nigeria.

Urine and faecal samples were obtained from 1,517 people in Fier, a typical rural village in Plateau State, Nigeria, for a parasitological survey among the population. 643 (42.39%) persons were found to be infected with altogether 9 helminths, namely: Ascaris lumbricoides, hookworm, Taenia sp., Trichuris trichiura, Enterobius vermicularis, Schistosoma mansoni, S. haematobium, Hymenolepis nana and Strongyloides stercoralis. Age and religion as opposed to sex, type of sewage system, and type of housing had a significant effect (P < 0.05) on the prevalence rates of the helminths in the population. Snail vector survey for schistosomatosis revealed the presence of Bulinus (Bulinus) truncatus, Bulinus (Physopsis) globosus and Biomphalaria pfeifferi with the latter being the most common with brevifurcate cercariae, and xiphidiocercariae being the most common cercariae harboured by the snails.


Trials to infect Anopheles stephensi with Plasmodium yoelii nigeriensis by the membrane feeding technique.

The aim of this study was to find optimal conditions for the membrane feeding technique to obtain maximum infection rates of mosquitoes with Plasmodium yoelii nigeriensis. The results show that the malaria parasite Plasmodium yoelii nigeriensis is most infective to Anopheles stephensi mosquitoes on day 3 of the infection in the mice, 1 day before the peak of parasitaemia. The mortality rate of the mosquitoes fed on mice on day 3 after infection was the highest as compared to mosquitoes fed on other days after infection. Gametocytes from mice 3 days after infection were fed to mosquitoes by three different membrane feeding methods. The results indicate that feeding during the first 10 min after blood collection gave the highest infection rates. Keeping the blood meal at a pH of 7.2 yields higher infection rates than keeping it at pH of 8.5. Stirring of the blood and supplying it with CO2 is not necessary when feeding of the mosquitoes is completed within the first 10 min after collection of the blood.


On the developmental velocity of Wucheria bancrofti larvae in vector mosquitoes of different susceptibility to filarial infections.

The developmental velocity of Wucheria bancrofti larvae was investigated in mosquito strains with different susceptibility to this filarial species. High susceptibility of the vector strains favoured fast filarial development, e.g. between 17.1 and 25.9% of all discovered larvae on day 13 p.i. had already migrated to the head and mouth parts of the more and partly even highly susceptible Anopheles gambiae, Culex pipiens quinquefasciatus and Aedes aegypti, respectively, whereas between 0 and 3.4% infective larvae had reached the head in the less susceptible Culex strains. On day 15 p.i. in all mosquito strains the majority of the larvae had reached infectivity.


[Morphological and biological characterization of a pure strain of Eimeria mitis].

A pure strain of Eimeria mitis was obtained from a crude field isolate by single oocyst inoculation of chicken. The strain was identified as Eimeria mitis by the morphology of the oocysts. Sporulated oocysts measured 16.1 +/- 2.1 microns in length and 13.6 +/- 1.0 microns in width. The shape index was 1.18. The prepatent period was 4 days and patency lasted 12 to 13 days. The highest number of oocysts was shed at day 6 after infection of chicken with 1 x 10(4), 5 x 10(4), or 1 x 10(5) oocysts and at day 8 after infection with 5 x 10(5) oocysts. A single infection of chicken with 1 x 10(4), 5 x 10(4), 1 x 10(5), or 5 x 10(5) oocysts resulted in reproduction values of 1:118,662, 1:15,720, 1:9,977 and 1:731, respectively.


Observations on mansonellosis among the Ibos of Abia and Imo States, Nigeria.

Between November, 1988 and April, 1991, parasitological and symptomatological methods of diagnosis were used to survey the prevalence of mansonellosis among the Ibo population in Abia and Imo States of Nigeria. 1,197 or 28.6% of the 4,183 persons examined were positive for microfilariae of Mansonella perstans. The prevalence of mansonellosis was significantly higher (P < 0.05) among rural dwellers (34.6%) than among urban dwellers (22.5%), in males (30.8%) than in females (26.3%), in farmers (59.8%) and palm wine tappers (46.1%) than in civil servants (7.6%), and in persons 21 years of age and above (36.2%) than in those in the first two decades of life (9.4%). Clinical signs observed in most infected persons include body itching, joint and back pains, occasional giddiness and elephantoid scrotum. Body itching was the most commonly observed clinical sign (14.7%), followed by joint pains (12.41%) with elephantoid scrotum (3.5%) as the least. The public health implication of the findings is discussed.


An orthodontic study of temporomandibular joint disorders. Part 1: Epidemiological research in Japanese 6-18 year olds.

Malocclusion is considered one of the etiological factors of temporomandibular joint disorder (TMD). The purpose of this study was to investigate the prevalence of TMD and the relationship between TMD and the type of occlusion. The sample consisted of 7337 Japanese children, 6-18 years old, 3219 boys and 4118 girls. TMD symptoms were recorded as well as the type of occlusion in children with TMD. The prevalence of TMD overall was 12.2%. The prevalence increased with age and was slightly higher in girls (13%) than in boys 11.1%. This difference was not statistically significant. Joint sound as the only symptom was more common in younger subjects. TMD symptoms seemed more complicated with age when pain and abnormal jaw movement combined with sound. Joint sound was the most common symptom (89.3%), followed by the combination of sound and pain (2.2%). The incidence of other symptoms was under 1%. In subjects with TMD, 24.9% exhibited crowding, 20.1% had excessive overjet, 6.8% deep bite, 6.3% edge-to-edge bite, 5.6% anterior crossbite, 5.4% open bite, and 3.8% posterior crossbite. Morphologically normal occlusion was observed in 27.1%. In this study, many subjects with TMD had malocclusions. Early treatment may be important in the prevention of severe TMD. Although those with morphologically normal occlusions were included, a more detailed study concerning other causes of TMD is needed also.


Blood flow changes in gingival tissues due to the displacement of teeth.

Changes in human gingival blood flow were measured using a Laser doppler flowmeter. The change of blood flow was correlated to the degree of force applied and there were variations in measurement of decreased blood flow among the subjects. The variation was attributed to the degree of tooth displacement and the size of the interdental space. This study examined the effect of tooth displacement on the gingival blood flow, as well as age and sex differences. Blood flow in gingival tissue was measured using a laser doppler flowmeter, and displacement of the maxillary incisors was measured using an eddy current sensor. The correlation coefficient of the decreased blood flow to the tooth displacement was 0.809, and it was higher than that to the degree of applied force (r = -0.625). The regression coefficient of decreased blood flow to the displacement of teeth was significantly correlated to the interdental space. The regression coefficient of decreased blood flow to the percentage of tooth displacement was independent of the interdental space. However, the regression coefficient of decreased blood flow to the percentage of tooth displacement was significantly higher in young subjects than in adults.


The effects of artificial saliva and topical fluoride treatments on the degradation of the elastic properties of orthodontic chains.

The effect of artificial saliva and topical fluoride treatments on the force relaxation and change in force delivery by three brands of elastomeric chains over a 4 week period was studied. The effect of storage in air and in the different test media on the distraction to achieve forces of 150g and 300g was determined for the chains. The effect of the test media on load relaxation of the chains was also examined. Elastomeric chains exhibit good elastic behavior when distracted to an initial force of less than 300g. When forces exceeded 300g, permanent deformation occurred and the force delivery was less predictable. Exposure to artificial saliva and topical fluoride affected the elastic properties of the elastomeric chains and increased the distraction required to deliver both the 150g and 300g force. The increase in distraction for a force of 150g, however, was relatively small and probably insignificant in the clinical setting. The distraction required to produce 300g was significantly larger and appeared to be clinically significant. Pre-stretching the elastomeric chains by 100% of their initial length was not found to be advantageous in terms of the load relaxation behavior. There was less load relaxation found in chains that were immersed in distilled water and Acidulated Phosphate Fluoride than in chains exposed only to air.


The influence of unilateral cleft lip and palate on maxillary dental arch morphology.

A sample of 97 untreated cleft lip and palate adult patients, with and without Simonart's band, was analyzed. The dimensions and form of the maxillary dental arches were analyzed. Comparison of this sample with a "normal" group indicated maxillary dental arch size and shape are distorted by the presence of a cleft which is characterized by a constriction that becomes more severe in the medial and anterior regions. The presence of Simonart's band affects the cleft arch form, redirecting the anterior extremity of the major segment towards the minor segment.


The asymmetric extraction decision.

Orthodontic study models, intraoral photographs, radiographs and diagnostic set-up casts of four completed adult cases are presented to show the possibilities of atypical treatment planning to correct specific and unusual malocclusions. The cases presented required asymmetric dental extractions in order to achieve the desired treatment results. Three diagnostic procedures were used to assist in making the final treatment decisions; a Bolton tooth size analysis, a space available/space needed assessment, and a pretreatment diagnostic set-up. The completed treatment results closely approximate the original diagnostic predictions, and completely satisfied each patient's chief concern. Most orthodontic practices now treat increasing numbers of non-growing (adult) patients. Traditional extraction considerations must be modified to satisfy the treatment needs of this demanding patient population.


TMJ biocompatible orthodontic treatment.

The papers summarized here indicate that TMJ dysfunction remains a complicated problem, requiring a multidisciplinary team approach. Psychological stress is an important factor in diagnosis. New concepts of joint function must be considered. The functional anatomy of the TMJ from an arthroscopic perspective should be studied. New treatment methods, such as the polycentric hinge joint articulator, should be considered. And finally, orthodontic diagnosis and treatment conventions need to be modified, from obtaining a complete history, clinical examination, arriving at a diagnosis and obtaining informed consent for treatment that may include psychological counseling, splint therapy, simultaneous fixed orthodontics and splint therapy and possible TMJ arthroscopic surgery for nearly all orthodontic patients.


Systemic sclerosis. The role of the mast cell and cytokines.

This review of SSc has addressed the clinical features, pathophysiology involving the mast cell, T cell cytokines, and their interaction with other inflammatory cells (Figs 4 and 5). Therapeutic dilemmas and the future use of immunomodulatory therapy is also discussed. The allergist-immunologist may be the first physician to identify this insidious condition, since patients may present with cutaneous changes, swelling, pruritus, dyspnea, and with obstructive and restrictive findings on pulmonary function testing, or gastrointestinal symptoms, mimicking gastrointestinal hypersensitivity.


Treatment of atopic dermatitis with alpha 1-proteinase inhibitor.

Alpha 1-proteinase inhibitor (alpha 1-PI), a serine protease inhibitor, was tested for its efficacy for the treatment of recalcitrant atopic dermatitis. Atopic dermatitis affects both children and adults and has no established etiology. We hypothesized that during inflammation there is an excess of serine proteases and a deficiency of their naturally occurring inhibitors at the local site of tissue injury, even though there is a normal serum level of serine protease inhibitors. This pilot study consisted of a nonblinded trial using alpha 1-PI at a concentration of 20 mg/mL in an aqueous solution in an alternate day schedule in conjunction with a 1% cream of alpha 1-PI (Stage I) and a 5% cream of alpha 1-PI for maintenance therapy (Stage II). Before enrollment in this trial all six patients failed to respond to high potency topical steroids. Safety was gauged by careful clinical monitoring of subjective complaints, objective findings of erythema, edema, and serial measurements of blood chemistries and complete blood counts. Wound healing was documented by serial photography. Written informed consent was obtained from each patient. All six patients showed significant clinical improvement within 6 to 21 days of initiation of alternate day therapy. Alpha 1-PI stopped pain, pruritus, and promoted tissue healing without scarring in all six patients. No adverse side effects of therapy were documented by clinical history, physical examination, or by blood studies after 120 days of therapy. Atopic dermatitis may be one example where inflammation is due to an imbalance of serine proteases and their naturally occurring inhibitors.(ABSTRACT TRUNCATED AT 250 WORDS)


Aerobiology of the Colorado Rockies: pollen count comparisons between Vail and Denver, Colorado.

Pollen patterns were compared between Vail, CO (8,200 feet elevation), Aspen, CO (7,900 feet) and Denver, CO (5,280 feet) from 1984 through 1988. Counts were obtained at all sites with a volumetric intermittent cycling rotating impaction sampler. Aspen and Denver were compared in 1984, and Vail and Denver from 1985 through 1988. While counts were generally lower in the mountain sites than Denver, certain pollens, especially trees, were quite high. Ragweed was essentially absent from Aspen and Vail, and chenopod-amaranth counts were very low. Cedar, pine, and aspen frequently pollinated despite active snowfall.


Measurement of IgG subclass antibodies to the group II antigen of Dermatophagoides pteronyssinus (Der p II) in sera from children with bronchial asthma.

Seven IgG-binding components in crude mite allergen extracted from Dermatophagoides pteronyssinus (Der p) were identified by immunoblotting analysis. Of these components, the 15-kDa protein was strongly recognized by IgG from patients with bronchial asthma, whereas it was less recognized by IgG from healthy individuals. Then, the 15-kDa protein was purified by reverse-phase high performance liquid chromatography and the protein was demonstrated to induce immediate positive skin reactions in patients with bronchial asthma. The N-terminal amino acid sequence of the 15-kDa protein was also determined and was in agreement with that of Der p II. In an attempt to understand the role of IgG subclass antibody of patients with bronchial asthma, we quantitated the antibodies of IgG subclass to Der p II. The levels of antibodies of total IgG and of all IgG subclasses in patients with bronchial asthma who had never received immunotherapy were higher than those of nonatopic healthy individuals. The levels of Der p II-specific IgG1 and IgG4 in asthmatic patients were 1.97 times and 2.20 times higher, respectively, than those in the control group. The present study demonstrates that patients with bronchial asthma are able to produce antibodies of all IgG subclasses specific to Der p II in response to natural exposure to mite antigen.


Clinical evaluation of the "Siriraj Spacer" in asthmatic Thai children.

We designed a new, washable, and collapsible bag spacer (the Siriraj Spacer) for use with metered-dose inhalers (MDI) by Thai asthmatic patients. The Siriraj Spacer consists of a mouthpiece, a front panel to which any type of MDI could be attached and a collapsible 800 mL ringed-plastic bag. Fifteen asthmatic children (6-13 years of age) were enrolled into a randomized, double-blind, triple crossover study (spanning a period of three days) to compare the clinical effectiveness of the Siriraj Spacer with that of the Volumatic Spacer (Glaxo, Inc., Research Triangle Park, NC, U.S.A.) and with the use of MDI alone. Medication used with the active method of administration was 2 puffs of albuterol (100 micrograms/puff) while 2 puffs from placebo MDI were used with the other two methods in succession. All children were stable asthmatic patients, and had been instructed how to use MDI properly by an open mouth technique just before the initiation of the study. Spirometry (FEV1, FVC, PEFR, and FEF25-75%) was followed for six hours after the administration of albuterol. The baseline FEV1s of the three study days were within 50% to 70% of the predicted values (with baseline variability of less than 20%). Data were expressed as percentage of improvement from baseline. By an analysis of variance with repeated measures, no significant differences were observed between pulmonary function data obtained with any of the three methods of bronchodilator administration (P > .05) at any time point throughout the 6-hour period.(ABSTRACT TRUNCATED AT 250 WORDS)


Buckwheat-induced anaphylaxis: a case report.

Buckwheat (Fagopyrum schulentum) is not taxonomically related to wheat and other cereal grains. Buckwheat flour is used as a wheat substitute in breads, biscuits, pancakes, and crepes. Occupational exposure to buckwheat flour has been associated with rhinitis, conjunctivitis, contact urticaria, and occupational asthma. We present a patient who developed urticaria and hypotension after ingestion of buckwheat crepes. Skin testing by the prick technique revealed 3+ positive reaction to buckwheat with negative reactions to other foods including wheat, egg white, and milk. RAST for anti-buckwheat IgE was strongly positive. Buckwheat ingestion is a potential cause of food-related anaphylaxis. There does not appear to be cross-reactivity between buckwheat and wheat allergy.


Evaluation of immune complexes after immunotherapy with wheat flour in bakers' asthma.

Inhalant food allergy has been described many times in literature, but double-blind clinical trials to support successful hyposensitization to these allergens has seldom been reported. Some authors have suspected that certain adverse reactions after immunotherapy may be mediated by immune complexes. Furthermore, the FDA does not recommend injection therapy with food extracts. We present a study on the detection of adverse effects after immunotherapy with an inhalant food (wheat flour) in a double-blind clinical trial in 26 patients with bakers' asthma. We investigated the presence of circulating immune complexes (CICs) after 2 years of treatment with hyposensitization to wheat flour.


Increased incidence of stings in venom-sensitive patients.

We compared the histories of 29 venom-sensitive and 28 control subjects who were selected from our venom referral and general allergy clinics respectively. The variables in the study included insect avoidance knowledge, the number of stings during the previous 2 years, insects involved, and time spent out of doors per week. There was no significant difference between the two groups with respect to age. All venom-sensitive patients were well versed in avoidance techniques while only 3 of 28 controls (11%) claimed such knowledge. Venom-sensitive subjects were stung almost ten times more frequently than control subjects. Wasp stings were the most common, followed by yellow jacket, honey bee, and hornet. The venom-sensitive patients also reported spending a greater amount of time outdoors (x 17.4 hours versus x 11.8, P < .05). An analysis of covariance showed that this difference in outdoor exposure was insufficient to account for the disparity in the number of stings. We conclude that other factors such as intrinsic attractants must be responsible for this phenomenon.


[Human pancreatic lipase activity: review of methods and general recommendations].

The properties of human pancreatic lipase are described, especially as regards the influence of the kind and the presentation of the substrate, as well as the effects of bile pigments and colipase. The authors have made a classification of the described methods for the determination of lipase activity in serum or plasma and have proposed recommendations for this assay.


[Serum myoglobin: evaluation of two immunolatex methods. Comparison with a radioimmunoassay method].

Latex immunoturbidimetric (ITA) and immunonephelemetric (INA) assays, were evaluated and compared to a radioimmunological assay (RIA) for the determination of serum myoglobin. Within-run and between-run assays and accuracy and linearity studies gave satisfactory results, very similar to those of RIA. Results were closely correlated with those of RIA for 100 samples of up to 2,000 micrograms/l (correlation coefficients respectively 0.96 and 0.97 for ITA and INA). Reference values (150 men and 150 women; range 20-65 years) were determined and expressed in percentiles. It is concluded that the two latex immunologic assays facilitate laboratory myoglobin evaluation (no radioactive material, short performance time). ITA is more suitable for emergency analysis and INA is more convenient for serial analysis and particular samples (presence of hemoglobin or chylomicrons).


[Comparative study of LpAI lipoparticles, HDL cholesterol and apolipoprotein AI in a control population, in a group of subjects with coronary diseases, and in a group of subjects with angiographically normal coronary vessels].

A new method for directly measuring LpAI lipoparticles containing apolipoprotein AI, but not apolipoprotein AII, is now disponible for laboratories. Concentrations of LpAI were measured in serum from 158 presumably healthy normolipidemic subjects (72 male, 86 female), for the age group 30-60 years. Concentrations of LpAI were also measured in subjects with angiographically defined coronary artery disease (coro+) and without angiographically defined coronary artery disease (coro-). After comparison of the groups, lipoprotein particle LpAI did not appear to be a better discriminative marker than HDL cholesterol or apolipoprotein AI for atherogen risk.


Spectrophotometric determination of lipase activity in the presence of increased triolein concentration.

A novel approach for the determination of pancreatic lipase (EC 3.1.1.3) activity by a spectrophotometric method is described. Enzyme activity is measured in the presence of 1 mmol/l triolein, a concentration much higher than usually employed in turbidimetry. The primary reaction medium is optimized as regards bile salt and colipase concentrations. The released fatty acids are enzymatically determined via a secondary reaction scheme using the constituents of a commercially available kit. The proposed method is easy to perform and may prove useful in determining lipase activity of standard lipase preparations, which is required for indirect assays. In addition to satisfactory precision and linearity as well as close correlation with other lipase assays, the procedure described in the present paper by-passes a number of short-comings (eg uncontrolled increase in absorbance) inherent to the turbidimetric methods.


Signal enhancement in fluorescence microscopy and flow cytometry using fluorescent liposome-antibody conjugates as second layer reagents.

We have developed small unilamellar liposomes (SUV) containing carboxyfluorescein for indirect immunophenotyping of cells by fluorescent microscopy and flow cytometry. Covalently coupled to anti-mouse Ig and anti-human Ig with mouse monoclonal antibodies and human typing antisera respectively, these conjugates can increase sensitivity by a factor of 10 compared to fluorescein isothiocyanate (FITC) conjugates. No increase in background fluorescence was observed. The reagents have been used to detect a low density antigen, the interleukin-2 receptor, on resting T-cells and to overcome the quenching effect of haemoglobin in fluorescent phenotyping of red cells. This has allowed to accurately estimate blood grouping in mixed fields and with small numbers of cells. We have determined the blood group of fetal red cells obtained from a chorionic villous sample at 10 weeks gestation and predicted severe hemolytic disease. This was confirmed by fetal sampling at 19 weeks and several intra-uterine transfusions were undertaken before a successful delivery at 36 weeks. Applications of the method in leucocyte phenotyping and transfusion medicine are discussed.


Biosensors: a new realism.

For many years biosensors have been hailed as the solution to many analytical problems. There is general agreement that biosensors offer the potential for easy-to-use, low-cost, rapid analysis. With such versatile, economic, reliable and cheap analytical devices at their disposal, manufacturers in industries as diverse as pharmaceuticals, food and drink, medical diagnostics and defence must surely be reaping vast profits from their biosensor-based products? In fact, biosensors have made only a very modest impact and this article attempts to present a realistic review of their current commercial potential. Consideration is given to the features and benefits of biosensors, the potential application markets, the impact of legislation, the needs of the user and the real commercial potential in the light of these factors and the existing competition.


Modification of the selected method for the determination of serum iron. Substitution of bathophenanthroline by ferene S.

The previously selected method (1977) for the measurement of iron in human serum has been modified by the working group on Iron and Iron-Transport Proteins (Sociéte Française de Biologie Clinique). Ferene S is used as the chromogen, it is more sensitive and cheaper than bathophenanthroline. The sample size has been reduced from 2 to 0.5 ml. No interference could be shown with high concentrations of bilirubin, copper or hemoglobin.


[Modified LDL and atherosclerosis. Nature of modifications. Physicochemical and biological properties].

The role of plasma LDL in atherogenesis is now well established. Cholesteryl ester accumulation within macrophages leads to foam cell formation, an early atherosclerotic process. In vitro, foam cell formation scarcely ever occurs in the presence of native LDL. Modification of these lipoproteins is necessary for their binding to macrophage scavenger receptors. In vitro modifications reported have involved chemical reactions, physical mechanisms, enzymatic reactions, cellular interactions and association with macromolecules. In vivo, they can occur by glycation or desialylation, smoking or by hemodynamic interactions. Alterations of the physicochemical properties of LDL are induced by oxidation and include an increase in their density and their net electronegative charge, changes in lecithin composition, polyunsaturated fatty acid peroxidation of lipids and apolipoprotein B100 degradation. Apolipoprotein B100 fragmentation leads to an impairment of uptake through LDL receptors, while uptake through macrophage scavenger receptors is enhanced. Modified LDL have also other particularities such as cytotoxicity, chemotactism for circulating monocytes, inhibition of resident macrophage mobility, vasoconstriction, perturbation of the arachidonic acid cascade, involvement in haemostasis and immune mechanisms. Hypotheses concerning the role of modified LDL, in particular oxidized LDL, in atherogenesis open new therapeutic prospects.


[LH and TSH receptors. A new family of G protein-coupled receptors].

Monoclonal antibodies have been raised against porcine LH receptor and allowed to clone the corresponding messenger RNA from testicular cells. Cross hybridisation with the LH receptor clone allowed to isolate a clone corresponding to the human TSH receptor from thyroids. The structure of both receptors have been determined. They show similarities but also differences to other G protein coupled receptors. In particular a large extracellular domain is specific of that new family of receptors. Variant forms of the LH receptor lacking transmembrane domains have been isolated. The obtention of monoclonal antibodies against both receptors allowed immunochemical and immunocytochemical studies to be performed. The human LH receptor gene have been localized to chromosome 2p21 and TSH receptor gene to chromosome 14q31. The complete organisation of the human TSH receptor gene has been determined.


Application of ECCLS guidelines to the analysis of Toxoplasma IgG and Rubella IgG using laboratory kits based on the Meia method.

The Meia method is an enzymo-immunoassay involving fluorometric detection, which is used in the Abbott IMx automatic analyzer. The purpose of this report was to analyse the Meia Toxoplasma gondii IgG antibody and Rubella IgG antibody assays, following ECCLS guidelines for the analysis of laboratory kits. The results showed that between-run imprecision for Rubella IgG was close to 15%; for Toxoplasma IgG the percentage was 13%. The mean recovery for Rubella IgG was 104% and 94% for Toxoplasma IgG. The carry-over for Rubella IgG was 0.64% and 0.26% for Toxoplasma IgG which, in both cases, was less than the analytical variability. Both Meia and Elisa showed a linear relationship in the analytical range of the method. Comparing Meia with the Elisa method, constant and proportional differences were found for IgG Rubella and proportional differences for IgG Toxoplasma. The Meia method has many positive analytical features to recommend and it can easily be used in a multidisciplinary laboratory, needing only a small number of serum samples.


[An automatic gas scanning device, technical improvement for obtaining a favourable atmosphere for in vitro culture of anaerobic bacteria].

All methods for growth of anaerobic bacteria on solid media depend on the elimination of atmospheric O2 through use of a palladium catalyst (Deoxo-Catalyst), active in presence of at least 5% H2 with resultant formation of water. Anaerobic chambers and jars are the two conventional methods employed. Both are based on the elimination of air by means of a pump and its replacement with gas from a cylinder (evacuation-replacement technique). An alternative chemical technique for use in anaerobic jars consists of adding internal gas-generating sachets. The former techniques are more efficient but the trend, particularly in the clinical laboratories, is to use the simpler chemical system that has two inconveniences: a slow establishment of anaerobiosis, and a high cost. We propose a new system that does not require a vacuum pump and consists in flushing anaerobic jars with a convenient gas mixture (H2, CO2, N2: 4.5; 5; 90.5 v/v) by means of an automaton regulating both time and gas flow. Gas-liquid chromatography analysis of the gas inside the jar shows a rapid elimination of gaseous O2, whose residual concentration is low enough to permit growth of all anaerobes of clinical interest, including those which are more O2-sensitive. Comparative qualitative and quantitative data obtained with all available techniques demonstrate the advantages of the new system.


[Pertinence of simultaneous measurements of pO2 and sO2 on ABL 510].

Blood tonometry is the only method to assess the accuracy of the pO2 determination on blood gas analyzers. ABL instruments by Radiometer were tested by two types of tonometry (film and bubble tonometry) and the validity of the algorithm for pO2 correction was analysed with these results. The role of the presentation of the specimen is also discussed. For the precision study on the ABL 510 analyzer, coefficients of variation for pO2 were < 0.37% and < 1.7% for within-run and day-to-day series respectively. pO2 accuracy was excellent. Linearity was verified between 0-620 mmHg (82.5 kPa), and interinstrument comparisons demonstrated a strict correlation with the Ciba-Corning 178 instrument. ABL 510 measures simultaneously oxygen saturation by spectrophotometry. Analytical results showed an acceptable level of imprecision, but the definition and the clinical significance of this parameter are ambiguous.


[Hemodialysis and ferritinemia].

Iron-deficiency is a common phenomenon in chronic renal diseases and haemodialysis patients, a treatment with iron or transfusions is always provided in an early preventive way; yet, an overload may appear. Serum ferritin, in spite of analytic variability, remains at the present time a good witness to appreciate patients' iron stores. Authors report the results obtained with four commercial reagents in healthy population and in haemodialysed children or adults. The comparison of results for this parameter shows that haemodialysed sera, with or without treatment, have the same behaviour with reagents as those of healthy subjects.


[Quality assurance or quality control. Lessons from an intercomparison circuit].

This article deals with the evaluation of the three year results of the French interlaboratory control for serum copper and zinc analysis. We pointed out some results interpreted on the basis of the existence of differences between the notions of quality control and total quality assurance. The main finding was a surprising change in the annual range in the final results of the laboratories. All 'good' laboratories in the first year showed a significant fall in their individual performance the second year. We can interpret that as a falling off of vigilance in relation with the self satisfaction after the publication of the first results. We concluded that good laboratory practices must be accompanied by a quality program not limited to an analytical control.


Epidermal growth factor-like activity in mares' milk.

Epidermal growth factor (EGF)-like activity was measured in mares' colostrum and milk by radioreceptor assay. Milk samples were collected from 22 mares 1 or more times during early lactation. Samples of colostrum were taken after parturition and before the foal first suckled (presuckle), within 6 hours after the foal first suckled (postsuckle), and on days 1, 2, 4, and 8 of lactation. In the 5 mares from which milk samples were obtained at each sampling time, presuckle colostral mean EGF-like activity (17.8 ng/ml) was greatest (P less than 0.05). The mean values for EGF-like activity at all other sampling times were not significantly different from each other (postsuckle colostrum, 9.7 ng/ml; day 1, 9.6 ng/ml; day 2, 8.5 ng/ml; day 4, 8.0 ng/ml; day 8, 7.8 ng/ml).


Ultrasonography of the urinary tract of female sheep.

We determined the position, dimensions, and structure of the kidneys, ureters, bladder, and urethra of 62 female sheep by use of ultrasonography. A 5.0-MHz convex transducer was placed over the right flank to examine the kidneys, and a 5.0 MHz-linear transducer was used to examine the bladder and urethra transrectally. All examinations were performed on sheep in standing position. The left kidney was 7.1 to 8.9 cm long, 3.4 to 5.5 cm wide, and 3.3 to 4.7 cm deep. Diameter of the parenchyma and renal sinus of the left kidney ranged between 1.1 and 1.9 cm and 1.1 and 2.0 cm, respectively. Circumference of the medullary pyramids varied between 2.1 and 3.3 cm. Similar ultrasonographic measurements were obtained for the right kidney. The diameter of the bladder varied between 0.3 and 6.9 cm in 96.8% of the sheep. The diameter of the bladder could not be determined in 32% of the sheep because it was > 10 cm, and, therefore, was beyond the penetration depth of the scanner. The only part of the urethra that could be ultrasonographically visualized was the internal urethral orifice. It had diameter between 0.1 and 0.2 cm. The ureters could not be ultrasonographically visualized in any of the sheep examined. The urinary tract of 8 sheep was examined 10 times within 2 weeks to examine whether measurements were reproducible. The interassay variation coefficient determined ranged from 3.1 to 31.8%, although for most variables, it ranged between 5 and 11%.(ABSTRACT TRUNCATED AT 250 WORDS)


Radiographically determined growth dynamics of primary lung tumors induced in dogs by inhalation of plutonium.

Beagles were exposed to aerosols of 239PuO2, 238PuO2, or 239Pu(NO3)4. Exponential growth constants for 50 primary lung tumors (23 bronchioloalveolar carcinomas, 22 papillary adenocarcinomas, 5 adenosquamous carcinomas) were calculated in 37 dogs, using sequential thoracic radiography. A wide range in doubling time (6 to 287 days) was observed. Mean +/- SEM doubling time was 93 +/- 10 days for bronchioloalveolar carcinoma, 107 +/- 13 days for papillary adenocarcinoma, and 101 +/- 36 days for adenosquamous carcinoma. Lung tumor growth rate in dogs was comparable to that in human patients with similar histologic tumor types. Linear regression analysis revealed significant (P < or = 0.0001) correlation between doubling time and survival of individual dogs. Doubling time was not significantly dependent on tumor type, sex, age at time of diagnosis, initial lung deposition, or isotope. Extrapolating time to tumor onset from tumor doubling time cannot be used to reliably predict the onset of malignancy.


Arterial blood gas tensions in healthy aged dogs.

Twenty-four healthy dogs > 8 years old were recruited. In each instance, arterial blood gas tensions were analyzed. The alveolar-to-arterial oxygen gradient (P[A-a]O2) was calculated to assess adequacy of pulmonary gas exchange. Thoracic radiographs were evaluated to ensure lack of visible signs of pulmonary disease and that lung features were similar to those in aged dogs of previous reports. Unlike findings in aged human beings, arterial partial pressure of oxygen (PaO2) was not decreased in this group of aged dogs (mean +/- SD, 102.9 +/- 7.8 mm of Hg). Similarly, P(A-a)O2 also was not increased. The thoracic radiographic findings were consistent with those of previous reports of pulmonary changes in aged dogs. The extent of radiographic abnormalities and the PaO2 were not correlated.


Effect of diet on results obtained by use of two commercial test kits for detection of occult blood in feces of dogs.

To evaluate the effect of diet on results obtained by use of 2 commercial test kits for detection of occult blood in feces, 5 dogs were fed 7 diets in randomized sequence. Dry and canned diets with various principal ingredients were evaluated. Each diet was offered twice over a 24-hour period, followed by a 36-hour nonfeeding period. Fecal specimens were collected twice daily, and tests for occult blood were performed within 12 hours. The dietary origin of fecal specimens was confirmed by use of colored markers fed with each diet, and was correlated with estimates of gastrointestinal tract transit time. A modified guaiac paper test and an o-tolidine tablet test were performed on each specimen. Of 59 specimens, 4 were positive for occult blood, using the o-tolidine tablet test. Three positive results were associated with a mutton-based canned diet, and 1 positive result was associated with a canned beef-based diet. Of 59 specimens, 11 were positive for occult blood, using the modified guaiac paper test. Four positive results were associated with the mutton diet, and 3 positive results were associated with the beef diet. Of the remaining 5 diets, 4 caused 1 positive reaction. Results were inconsistent with the null hypothesis that the distribution of positive occult blood test results is not affected by diet (P < 0.025), and indicate that diet can affect the specificity of peroxidase-based tests for detection of occult blood in canine feces. Diet modification prior to testing is recommended.


Determination of protein concentrations and their molecular weight in tears from cats with normal corneas and cats with corneal sequestrum.

Protein concentration was determined, using the Bradford technique, in tears from cats with normal corneas and from cats with corneal sequestrum. Tears from the former group contained 5.81 +/- 2.29 mg of protein/ml; those from corneal sequestrum-affected cats contained 6.21 +/- 2.21 mg/ml. Difference between the 2 values was not significant. Molecular weight determination was made, using 4 to 20% sodium dodecyl sulfate-polyacrylamide gels. Molecular mass of proteins ranged from 263 to 14 kDa. There was no detectable difference in the band patterns for the 2 groups.


Evaluation of an automated system for hemoglobin measurement in animals.

In veterinary medicine, PCV determined by centrifugation of blood in a microhematocrit tube is the most common clinical test used to initially assess and monitor anemic and polycythemic animals. In contrast, blood hemoglobin (Hb) concentration, rather than PCV, is generally determined in human patients. One automated system photometrically measures blood Hb concentration after conversion of Hb to azide methemoglobin without dilution and was found to be a simple and accurate instrument for use in human medicine. We evaluated the system for its accuracy in measuring blood Hb concentration in animals by comparing it with standard techniques and for its suitability in veterinary practice. Blood samples, anticoagulated with potassium EDTA, from 78 healthy animals (33 dogs, 17 cats, 13 horses, and 15 cows) and 58 dogs and 4 cats with various blood abnormalities (10 anemia, 11 polycythemia, 21 lipemia, 16 leukocytosis, and 6 icterus) were analyzed. In all species, blood Hb concentration of healthy animals determined by the system was comparable to that measured by standard cyanmethemoglobin methods (ie, an automated counter; rI = 0.987 to 0.998 and a hemoglobin kit, rI = 0.946 to 0.993). The aforementioned system also yielded similar values to those obtained by use of standard methods in anemic, polycythemic, and icteric dogs and cats. Moreover, the system reads the absorbance at 2 wavelengths to correct for turbidity, and therefore, accurately measured Hb concentration in blood samples with severe lipemia (triglycerides concentration > 500 mg/dl) and marked leukocytosis (> 50,000 WBC/microliter), whereas other standard Hb techniques are known to give falsely high results.(ABSTRACT TRUNCATED AT 250 WORDS)


Reference serum biochemical values for emus and ostriches.

Reference serum biochemical values were determined in blood samples from 15 male, 18 female, and 4 unsexed emus (Dromaius novaehollandiae) 1 to 48 months old. Serum biochemical values also were obtained for 19 male, 26 female, and 4 unsexed ostriches (Struthio camelus) 1 to 60 months old. Parametric (mean +/- 2 SD) and non-parametric (fifth to 95th percentile) reference ranges and linear trends as influenced by age were determined for enzyme activities and concentrations of glucose, inorganic phosphate, BUN, uric acid, creatinine, triglyceride, cholesterol, total protein, and albumin. Species differences for all analytes, except cholesterol and inorganic phosphate concentrations, were detected. Creatine kinase values in ostriches were higher than those in emus. There were no linear relationships between age and analyte values in emus, and sex did not significantly (P < 0.05) affect the values in emus. Analyte values in ostriches tended to increase with age, but cholesterol, creatine kinase, inorganic phosphate, and alkaline phosphatase concentrations decreased with age. Glucose, triglyceride, gamma-glutamyltransferase, and cholinesterase concentrations in ostriches were not linearly associated with age. Age had a greater effect on the analyte values of female ostriches than it did on male ostriches. Concentrations generally increased with age in female ostriches, except for cholesterol, cholinesterase, inorganic phosphate, and alkaline phosphatase concentrations, which decreased with age.


Furosemide-induced electrolyte depletion associated with echinocytosis in horses.

Echinocytes have been incriminated in the pathogenesis of exertional diseases in horses. To evaluate the hypothesis that echinocytes are dehydrated erythrocytes, we decreased blood sodium and potassium concentrations in 4 horses by administering furosemide (1.0 mg/kg of body weight, q 12 h) for 2 days and we monitored CBC, serum and erythrocyte sodium and potassium concentrations, and echinocyte numbers. Serum sodium concentration decreased progressively over the 48 hours of furosemide administration, then returned to near baseline concentration at 168 hours. A statistically significant decrease (P < 0.05) in serum potassium concentration was observed at 24, 48, and 72 hours after initial furosemide administration, and remained less than the baseline value at the end of the study. Mean erythrocyte potassium concentration decreased rapidly and remained low at the end of the study. Minimal changes were observed in erythrocyte sodium concentration during the first 72 hours after furosemide administration, but the value was significantly (P < 0.05) increased at 168 hours. Type-I and type-II echinocyte numbers increased by 4 hours after furosemide administration and persisted throughout the study. Type-III echinocytes were not seen in baseline samples, but numbers increased only modestly after furosemide administration. Administration of epinephrine to well-hydrated horses increased echinocyte numbers only minimally, indicating that splenic contraction was not the likely cause for the furosemide-associated increase. To determine whether the decrease in erythrocyte potassium concentration and increase in sodium concentration was caused by furosemide acting directly on the erythrocyte membrane, we quantified erythrocyte potassium and sodium concentrations before and after incubation with furosemide in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)


Reference hematologic values and morphologic features of blood cells in healthy adult llamas.

Hematologic values and cellular morphologic features were evaluated for 38 healthy adult llamas. Reference ranges were determined for PCV, reticulocyte concentration, leukocyte concentration, and leukocyte differential counts. The approach used in this study was to focus on hematologic values that may be determined by use of techniques readily available to the practicing veterinarian and nonveterinary laboratory. Unique cellular morphologic features commonly observed and interpreted as normal included large granular lymphocytes, hyposegmented eosinophil nuclei, folded erythrocytes, and hemoglobin crystals.


Characterization of erythrocytic indices and serum iron values in healthy llamas.

An electronic particle counter with attached particle-size analyzer was configured to directly determine concentration, mean cell volume, and volume distribution of erythrocytes in llama blood. Blood from 38 healthy llamas was used to characterize erythrocytic measurements and serum iron values for this species. Volume distribution curves for llama erythrocytes were similar in shape to those of other species. These curves had a unimodal, symmetric shape with a tail skewed to the right. Reference ranges for directly measured mean cell volume, erythrocyte concentration, hemoglobin concentration, and mean cell hemoglobin concentration were 21 to 28 fl, 11.3 x to 17.5 x 10(6) cells/microliters, 12.8 to 17.6 g/dl, and 43.2 to 46.6 g/dl, respectively. Reference ranges for serum iron concentration, total iron-binding capacity, and transferrin saturation were determined to be 70 to 148 micrograms/dl, 230 to 370 micrograms/dl, and 22 to 50%, respectively.


Variability of serum bile acid concentrations over time in dairy cattle, and effect of feed deprivation on the variability.

Twelve nonlactating dairy cows, free of signs of liver disease and with normal serum activities of liver-derived enzymes and normal liver biopsy tissue, were examined over a 72-hour period for serum total bile acid concentrations. The cattle were fed hay twice daily, and blood samples were obtained every hour for 24 hours, every other hour for 24 hours, then every hour for 24 hours. After 3 weeks, the study was repeated on 6 of the cattle, thus providing data for eighteen 72-hour periods. Serum bile acid concentration varied greatly over the 72 hours, with the range being from one third to 3 times the median. There were variations by as much as 60 mumol/L from 1 hour to the next. After another 3 weeks, 8 of the cattle were deprived of hay for 48 hours and then fed hay morning and afternoon of the third (last) day of the study. There was no significant reduction in bile acid concentration after withholding the hay, but the variability was reduced (P = 0.02) during the last 20 hours of the hay-deprivation period. In 3 ancillary studies, serum bile acid concentrations were examined over a 48-hour period in 2 cows in early lactation, 3 cows in midlactation, and two 6-month-old heifers. The cows were fed hay and grain twice daily, and the heifers were fed only hay twice daily. In comparison with values for the 12 nonlactating cows fed hay twice daily, mean serum bile acid concentration in the recently freshened cows was significantly (P < 0.002) higher (62.9 vs 22.0 mumol/L).(ABSTRACT TRUNCATED AT 250 WORDS)


Serum bile acid concentrations in clinically normal cattle: comparison by type, age, and stage of lactation.

Serum total bile acid concentrations were determined for various types and ages of cattle. There was extreme variability among all the cattle, but the variance was twice as large (0.50 vs 0.22 in logarithmic scale) for beef cattle than for dairy cattle. There was no significant difference in serum total bile acid concentrations between beef cattle and dairy cattle in midlactation. Values for calves < 6 weeks old and for 6-month-old heifers were significantly (P = < 0.05) lower than values for lactating dairy cows. The 5th to 95th percentile range of values (mumol/L) for beef cattle was 9 to 126; for lactating dairy cattle, 15 to 88; and for 6-month-old dairy heifers, 11 to 64.


Evaluation of hemostatic analytes after use of hypertonic saline solution combined with colloids for resuscitation of dogs with hypovolemia.

The effects of hypertonic saline solution (HTSS) combined with colloids on hemostatic analytes were studied in 15 dogs. The analytes evaluated included platelet counts, one-stage prothrombin time, activated partial thromboplastin time, von Willebrand's factor antigen (vWf:Ag), and buccal mucosa bleeding times. The dogs were anesthetized, and jugular phlebotomy was used to induced hypovolemia (mean arterial blood pressure = 50 mm of Hg). Treatment dogs (n = 12) were resuscitated by infusion (6 ml/kg of body weight) of 1 of 3 solutions: HTSS combined with 6% dextran 70, 6% hetastarch, or 10% pentastarch. The control dogs (n = 3) were autotransfused. Hemostatic analytes were evaluated prior to induction of hypovolemia (baseline) and then after resuscitation (after 30 minutes of sustained hypovolemia) at 0.25, 0.5, 1, 6 and 24 hours. All treatment dogs responded rapidly and dramatically to resuscitation with hypertonic solutions. Clinically apparent hemostatic defects (epistaxis, petechiae, hematoma) were not observed in any dog. All coagulation variables evaluated, with the exception of vWf:Ag, remained within reference ranges over the 24-hour period. The vWf:Ag values were not statistically different than values from control dogs, and actual values were only slightly lower than reference ranges. Significant (P < or = 0.04) differences were detected for one-stage prothrombin time, but did not exceed reference ranges. The results of this study suggested that small volume HTSS/colloid solutions do not cause significant alterations in hemostatic analytes and should be considered for initial treatment of hypovolemic or hemorrhagic shock.


Evaluation of serum fructosamine concentration as an index of blood glucose control in cats with diabetes mellitus.

Fructosamine, a glycated serum protein, was evaluated as an index of glycemic control in normal and diabetic cats. Fructosamine was determined manually by use of a modification of an automated method. The within-run precision was 2.4 to 3.2%, and the day-to-day precision was 2.7 to 3.1%. Fructosamine was found to be stable in serum samples stored for 1 week at 4 C and for 2 weeks at -20 C. The reference range for serum fructosamine concentration in 31 clinically normal colony cats was 2.19 to 3.47 mmol/L (mean, 2.83 +/- 0.32 mmol/L). In 27 samples from 16 cats with poorly controlled diabetes mellitus, the range for fructosamine concentration was 3.04 to 8.83 mmol/L (mean, 5.93 +/- 1.35 mmol/L). Fructosamine concentration was directly and highly correlated to blood glucose concentration. Fructosamine concentration also remained high in consort with increased blood glucose concentration in cats with poorly controlled diabetes mellitus over extended periods. It is concluded that measurement of serum fructosamine concentration can be a valuable adjunct to blood glucose monitoring to evaluate glycemic control in diabetic cats. The question of whether fructosamine can replace glucose for monitoring control of diabetes mellitus requires further study.


Effects of ketamine infusion on halothane minimal alveolar concentration in horses.

Eight adult horses were used in a study to determine ketamine's ability to reduce halothane requirement. To obtain steady-state plasma concentrations of 0.5, 1.0, 2.0, 4.0, and 8.0 micrograms/ml, loading doses and constant infusions for ketamine were calculated for each horse on the basis of data from other studies in which the pharmacokinetic properties of ketamine were investigated. Blood samples for determination of plasma ketamine concentrations were collected periodically during each experiment. Plasma ketamine concentrations were determined by capillary gas chromatography/mass spectrometry under electron-impact ionization conditions, using lidocaine as the internal standard. Halothane minimal alveolar concentration (MAC; concentration at which half the horses moved in response to an electrical stimulus) and plasma ketamine concentration were determined after steady-state concentrations of each ketamine infusion had been reached. Plasma ketamine concentrations > 1.0 microgram/ml decreased halothane MAC. The degree of MAC reduction was correlated directly with the square root of the plasma ketamine concentration, reaching a maximum of 37% reduction at a plasma ketamine concentration of 10.8 +/- 2.7 micrograms/ml. Heart rate, mean arterial blood pressure, and the rate of increase of right ventricular pressure did not change with increasing plasma ketamine concentration and halothane MAC reduction. Cardiac output increased significantly during ketamine infusions and halothane MAC reduction. Our findings suggest that plasma ketamine concentrations > 1.0 micron/ml reduce halothane MAC and produce beneficial hemodynamic effects.


Pharmacokinetics of metronidazole and its concentration in body fluids and endometrial tissues of mares.

Serum concentrations of metronidazole were determined in 6 healthy adult mares after a single IV injection of metronidazole (15 mg/kg of body weight). The mean elimination rate (K) was 0.23 h-1, and the mean elimination half-life (t1/2) was 3.1 hours. The apparent volume of distribution at steady state was 0.69 L/kg, and the clearance was 168 ml/h/kg. Each mare was then given a loading dose (15 mg/kg) of metronidazole at time 0, followed by 4 maintenance doses (7.5 mg/kg, q 6 h) by nasogastric tube. Metronidazole concentrations were measured in serial samples of serum, synovia, peritoneal fluid, and urine. Metronidazole concentrations in CSF and endometrial tissues were measured after the fourth maintenance dose. The highest mean concentration in serum was 13.9 +/- 2.18 micrograms/ml at 40 minutes after the loading dose (time 0). The highest mean synovial and peritoneal fluid concentrations were 8.9 +/- 1.31 micrograms/ml and 12.8 +/- 3.21 micrograms/ml, respectively, 2 hours after the loading dose. The lowest mean trough concentration in urine was 32 micrograms/ml. Mean concentration of metronidazole in CSF was 4.3 +/- 2.51 micrograms/ml and the mean concentration in endometrial tissues was 0.9 +/- 0.48 micrograms/g at 3 hours after the fourth maintenance dose. Two mares hospitalized for treatment of bacterial pleuropneumonia were given metronidazole (15.0 mg/kg, PO, initially then 7.5 mg/kg, PO, q 6 h), while concurrently receiving gentamicin, potassium penicillin, and flunixin meglumine IV. Metronidazole pharmacokinetics and serum concentrations in the sick mares were similar to those obtained in the healthy mares.


Effects of xylazine on airway function in ponies with recurrent airway obstruction.

The effect of IV administration of the alpha 2-adrenoceptor agonist xylazine hydrochloride (0.5 mg/kg of body weight) was examined in ponies with recurrent obstructive pulmonary disease, commonly called heaves. Six ponies with the disease (principals) were studied during clinical remission and during an acute attack of airway obstruction precipitated by stabling and feeding of dusty hay. Six control ponies were also studied. In principal ponies with airway obstruction, xylazine administration significantly (P < 0.05) decreased pulmonary resistance and increased dynamic compliance, but did not affect PaO2 or PaCO2. The alpha 2-antagonist yohimbine blocked the pulmonary effects of xylazine. Administration of saline solution was without effect in both groups of ponies at all periods and xylazine did not have effect in controls or in principals in clinical remission.


Effect of the somatostatin analogue octreotide on gastric fluid pH in ponies.

The effect of the somatostatin analogue, octreotide, on gastric fluid pH was investigated in 4 ponies. Gastric fluid pH was determined after SC administration of octreotide or physiologic saline solution (control). A baseline sample of fluid was obtained, the agent was given, and 8 additional samples were collected hourly. Administration of octreotide at all dosages tested (0.1, 0.5, 1.0, and 5.0 micrograms/kg of body weight) increased gastric pH to > 5.0. Baseline values were consistently < 2.7. Administration of octreotide at these same dosages induced gastric pH values > 4.0 for 2.4 +/- 1.2, 4.8 +/- 0.8, 5.7 +/- 1.3, and 5.4 +/- 2.6 (mean +/- SD) continuous hours, respectively. Treatment at all dosages increased the pH of gastric fluid, compared with control values. The duration of the increase in pH was significantly (P < 0.05) different than that of the control treatment, even for the lowest dosage, 0.1 microgram/kg.


Potential use of simple manganese salts as antioxidant drugs in horses.

The scavenging of superoxide radicals by endogenous and therapeutically administered superoxide dismutases may prevent superoxide-mediated oxidative stress leading to lipid peroxidation, membrane lysis, and cell death in a wide variety of normal and pathologic states. Simple inorganic manganous salts such as MnCl2 also have superoxide dismutase-like activity and are extremely inexpensive, compared with enzymatic superoxide dismutase preparations. In this study, we explored the use of Mn salts as antioxidant drugs. We used the percentage of inhibition of nitroblue tetrazolium reduction by superoxide as a measure of the amount of superoxide dismutase-like activity. We found concentration-related increases in superoxide scavenging activity in simple buffer solutions upon addition of 1.25, 2.5, and 5.0 microM MnSO4. To determine whether Mn salts can inhibit oxidative damage in tissues, we used an in vitro model of lipid peroxidation in ischemic and reoxygenated rat liver slices. Concentrations of 10, 100, and 1000 mumoles MnCl2/L of buffer significantly decreased indicators of lipid peroxidation believed to be initiated by intracellular superoxide. We then determined the effectiveness of MnCl2 as a superoxide scavenger in conscious horses by measuring the superoxide scavenging ability of equine plasma before and during intravenous infusions of 1.0 L volumes of 0.9% saline solution containing 0, 12.5, or 25 mM MnCl2. Plasma Mn concentrations, which were determined by atomic absorption spectrophotometry, increased as a function of time and dose. Intravenously administered MnCl2 concomitantly produced dose-related increases in superoxide scavenging ability of equine plasma at 15, 30, 45, and 60 minutes after the onset of infusion, compared with preinfusion control values.(ABSTRACT TRUNCATED AT 250 WORDS)


Comparative aspects and sex differentiation of plasma sulfamethazine elimination and metabolite formation in rats, rabbits, dwarf goats, and cattle.

Plasma disposition and urinary recovery of sulfamethazine (SMZ), its N4-acetylated metabolite (N4AcSMZ), and 2 of its hydroxylated metabolites--5-hydroxysulfamethazine (5OHSMZ) and 6-hydroxymethylsulfamethazine (6CH2OHSMZ)--were determined in either sex of 4 animal species: rats, dwarf goats, rabbits, and cattle. Rats, rabbits, and dwarf goats had significant (P < 0.01) sex difference in SMZ plasma clearance. Male rats had higher plasma clearance than did female rats, and excreted higher amounts of the hydroxy metabolites and lower amounts of N4AcSMZ. The N4AcSMZ metabolite was predominant in plasma and urine of rabbits. Male rabbits had higher plasma clearance than did female rabbits, but differences in metabolite profile were not apparent. With regard to plasma SMZ elimination, the situation in goats was opposite to that in rats. Male goats had considerably lower clearance than did female goats. This was associated with a lower hydroxylation rat in males. Plasma half-life of SMZ in cows was lower than that in bulls, probably because of a smaller distribution volume in cows. Compared with elimination via urine, elimination via milk was negligible in cows. Significant differences in metabolite profiles were not found between bulls and cows. Similar to those in rats and mice, hormone-dependent xenobiotic metabolic pathways may exist in other species. Depending on species and xenobiotic compound residue concentrations of xenobiotics, their metabolites, or both may differ with sex of the animal, or may be altered after treatment with anabolic hormones.


Effects of intramuscular administration of glycosaminoglycan polysulfates on signs of incipient hip dysplasia in growing pups.

We tested the hypothesis that treatment of growing, susceptible (to hip dysplasia) pups by IM administration of glycosaminoglycan polysulfates would mitigate the signs of incipient hip dysplasia. In 1 experiment, 7 pups, selected at random from 2 litters, were administered glycosaminoglycan polysulfates (2.5 mg/kg of body weight, IM) twice weekly, and 7 control pups from the same litters were given sterile buffered 0.9% saline solution from the age of 6 weeks to 8 months. Hip joints were examined by radiography, with pups in the standard, limbs-extended position. At 8 months of age, all pups in this experiment did not manifest femoral head subluxation radiographically. The Norberg angle, a measure of coxofemoral congruity, improved from a mean +/- SEM value of 102 degrees +/- 1 degrees in controls to 106 degrees +/- 1 degrees in treated pups (P = 0.008). Pups were not subjected to necropsy. In the second experiment, 8 pups were selected at random from 2 litters and were administered 5 mg of glycosaminoglycan polysulfates/kg, IM, twice weekly from 6 weeks to 8 months of age. Similarly, 8 control pups were administered saline solution. At 8 months of age, hip joints were examined by radiography with pups in the standard position; at necropsy, intra-articular tissues were evaluated macroscopically and biochemically. Of 8 treated pups, none had subluxation radiographically, whereas 4 of 8 control dogs had femoral head subluxation. Mean Norberg angle on the radiographs was 109.7 degrees +/- 1.6 degrees for the treated group and was 101.5 degrees +/- 1.6 degrees for controls, representing a mean improvement in coxofemoral congruity of 8.2 degrees in the treated pups.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of hypertonic and isotonic saline solutions on plasma constituents of conscious horses.

Blood constituents and vascular volume indices were determined in 5 standing horses by use of 2-period crossover experimental design. Horses were either administered hypertonic (2,400 mosm/kg of body weight, i.v.) or isotonic (300 mosm/kg, i.v.) saline solution. Each solution was administered at a dosage of 5 ml/kg (infusion rate, 80 ml/min). Samples for determination of PCV, plasma volume, blood volume, plasma osmolality, total amount of plasma protein and plasma concentrations of protein, Na, K, and Cl were collected at 0 hour (baseline, before fluid infusion) and 0.5 hour (at the end of fluid infusion), and subsequently, at 0.25- or 0.5-hour intervals for 4.5 hours. All horses were given the predetermined dose of fluids by 0.5 hour after beginning the saline infusion. Values of P < or = 0.05 were considered significant. Administration of hypertonic saline solution was associated with decreased mean body weight by 4.5 hours, but weight change after isotonic saline administration was not significant. Other than body weight and plasma protein concentration, between-trial difference (treatment effect) was not observed for any measured variable or index. The F values indicated that increasing the number of horses would have not changed these results. A time effect was evident across both trials, so that mean (+/- SD) plasma volume increased (12.3 +/- 1.07%) and mean plasma protein concentration (-12.1 +/- 1.03%) and PCV (-11.9 + 0.67%) decreased proportionately and transiently in association with administration of either fluid at that volume. Other time effects included increased plasma osmolality and Na and Cl concentrations. Blood volume estimates and total amount of plasma protein remained unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)


Hemodynamic and respiratory responses to variable arterial partial pressure of oxygen in halothane-anesthetized horses during spontaneous and controlled ventilation.

Cardiovascular and respiratory responses to variable PaO2 were measured in 6 horses anesthetized only with halothane during spontaneous (SV) and controlled (CV) ventilation. The minimal alveolar concentration (MAC) for halothane in oxygen was determined in each spontaneously breathing horse prior to establishing PaO2 study conditions--mean +/- SEM, 0.95 +/- 0.03 vol%. The PaO2 conditions of > 250, 120, 80, and 50 mm of Hg were studied in each horse anesthetized at 1.2 MAC of halothane and positioned in left lateral recumbency. In response to a decrease in PaO2, total peripheral resistance and systolic and diastolic arterial blood pressure decreased (P < 0.05) during SV. Cardiac output tended to increase because heart rate increased (P < 0.05) during these same conditions. During CV, cardiovascular function was usually less than it was at comparable PaO2 during SV (P < 0.05). Heart rate, cardiac output, and left ventricular work increased (P < 0.05) in response to a decrease in PaO2, whereas total peripheral resistance decreased (P < 0.05). During SV, cardiac output and stroke volume increased and arterial blood pressure and total peripheral resistance decreased with duration of anesthesia at PaO2 > 250 mm of Hg. During SV, minute expired volume increased (P < 0.05) because respiratory frequency tended to increase as PaO2 decreased. Decrease in PaCO2 (P < 0.05) also accompanied these respiratory changes. Although oxygen utilization was nearly constant over all treatment periods, oxygen delivery decreased (P < 0.05) with decrease in PaO2, and was less (P < 0.05) during CV, compared with SV, for comparable PaO2 values.(ABSTRACT TRUNCATED AT 250 WORDS)


Arteriovenous differences for glutamine in the equine gastrointestinal tract.

Glutamine has been shown to be an important metabolic substrate of enterocytes in many animals, including cats, dogs, hamsters, human beings, monkeys, rabbits, rats, and sheep. To determine whether glutamine is important in the metabolism of cells of the equine gastrointestinal tract, we examined transintestinal differences in glutamine concentrations in the arterial and venous circulation, and measured activity of the major glutamine catabolizing enzyme, glutaminase. Arteriovenous differences provide an index of the amount of a given substrate removed by the tissue across which the measurements are made, and commonly are expressed as a percentage of substrate removed, or percent extraction. Arteriovenous differences for glutamine were determined in 7 anesthetized adult horses (weight, 450 to 500 kg) before and after an i.v. glutamine infusion. The mean baseline arterial glutamine concentration (+/- SEM) was 572 +/- 24 microM; this concentration quadrupled (to 2,167 +/- 135 microM, P less than 0.01) 1 minute after i.v. bolus infusion of a 17.5-g glutamine load. Baseline extraction by the portal-drained viscera was 7.5 +/- 1.5%; this value increased to 18 +/- 2% at 1 minute (P less than 0.01) and had returned to baseline values 60 minutes later. Arteriovenous differences were greatest across the jejunum (11.8 +/- 1.8% in the baseline period vs 33.1 +/- 3.1% at 1 minute, P less than 0.001), with smaller differences across the colon, suggesting that the jejunum was the more avid utilizer of glutamine. Glutaminase activity was 4.38 +/- 0.16 and 4.00 +/- 0.60 mumol/mg of protein/h under standard conditions in jejunal and ileal mucosa, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Regional brain blood flow during prolonged submaximal exercise in ponies.

Experiments were carried out on 8 healthy ponies to examine the effects of prolonged submaximal exercise on regional distribution of brain blood flow. Brain blood flow was ascertained by use of 15-microns-diameter radionuclide-labeled microspheres injected into the left ventricle. The reference blood was withdrawn from the thoracic aorta at a constant rate of 21.0 ml/min. Hemodynamic data were obtained with the ponies at rest (control), and at 5, 15, and 26 minutes of exercise performed at a speed setting of 13 mph on a treadmill with a fixed incline of 7%. Exercise lasted for 30 minutes and was carried out at an ambient temperature of 20 C. Heart rate, mean arterial pressure, and core temperature increased significantly with exercise. With the ponies at rest, a marked heterogeneity of perfusion was observed within the brain; the cerebral, as well as cerebellar gray matter, had greater blood flow than in the respective white matter, and a gradually decreasing gradient of blood flow existed from thalamus-hypothalamus to medulla. This pattern of perfusion heterogeneity was preserved during exercise. Regional brain blood flow at 5 and 15 minutes of exercise remained similar to resting values. However, at 26 minutes of exercise, vasoconstriction resulted in a significant reduction in blood flow to all cerebral and brain-stem regions. In the cerebellum, the gray matter blood flow and vascular resistance remained near control values even at 26 minutes of exercise. Vasoconstriction in various regions of the cerebrum and brainstem at 26 minutes of exertion may have occurred in response to exercise-induced hypocapnia, arterial hypertension, and/or sympathetic neural activation.


Temporal lectin histochemical characterization of porcine small intestine.

A variety of biotinylated lectins was applied to formalin-fixed intestinal sections from isolator-reared pigs ranging in age from newborn through 12 weeks. Lectin binding to brush borders of villus enterocytes, crypt enterocytes, and dome epithelium, and lectin reactivity within goblet cells and Brunner's glands was semiquantified by microscopy and was used to estimate temporal changes in complex carbohydrates of enteric epithelium. Although variability in binding scores often was observed among pigs of the same age, several general patterns of lectin binding were detected. Dolichos biflorus and Ulex europaeus lectins had increasing binding to brush border membranes as pigs aged. The Dolichos biflorus, however, had decreased binding at the 12-week time point. Neuraminidase-treated Arachis hypogaea and Triticum vulgaris were associated with high mean binding scores at all time points. Canavalia ensiformis bound, with high mean score at all time points, to villus but not to crypt enterocytes. Arachis hypogaea was associated with variable but often high binding scores, regardless of pig age. Succinylated wheat germ agglutinin bound more to crypt than to villus enterocytes. Goblet cells were generally less reactive than were corresponding villi and crypts. Dome epithelium reactivity varied with the lectin used, whereas Brunner's glands reacted with all lectins tested. We conclude that age and regional variations in lectin binding may reflect differences in intestinal function and differentiation. Because complex carbohydrates may act as cell surface receptors for a variety of enteric pathogens, our results indicate that these differences may be partially responsible for age and anatomic differences in susceptibility or resistance to enteric disease.


Bacterial survival, lymph node changes, and immunologic responses of cattle vaccinated with standard and mutant strains of Brucella abortus.

Forty-eight cattle were used in 4 experiments; 6-week-old calves in experiments 1-3 (n = 24) and 10-month-old heifers in experiment 4 (n = 24). In experiments 1-3, 7 groups of 3 calves each were inoculated SC with 5 strains of Brucella abortus: virulent strain 2308 (2 groups), vaccine strain 19 (2 groups), and mutant strains RB51. 19 delta 31K, and 19 delta SOD. Sera and lymph node tissues were examined at 2-week intervals for evidence of infection. At postinoculation (PI) week 12, 2 calves in each group were given dexamethasone for 5 days. Calves were then euthanatized and lymphoid tissue, spleen, liver, and bone marrow were examined for evidence of B abortus. Calves given strain 2308 had large numbers of bacteria in their lymph nodes, marked granulomatous lymphadenitis in the deep cortex, and loss of lymphoid cells in superficial cortical areas. In addition, they had high serum antibody titers at PI week 16. Calves given strain 19, or genetic mutants derived from strain 19, cleared bacteria from lymph nodes more rapidly, had less lymphoid destruction, and developed antibody titers that did not persist for 16 weeks. The RB51 strain (rough) was cleared most rapidly from lymphoid tissues and induced serum antibody responses only to the core of the lipopolysaccharide molecule. Treatment of calves with dexamethasone did not cause B abortus to reappear in tissues of any calves, nor did serum antibody titers increase.(ABSTRACT TRUNCATED AT 250 WORDS)


Systemic and pulmonary antibody responses of calves to Pasteurella haemolytica after intrapulmonary inoculation.

Systemic and pulmonary antibody responses of calves to Pasteurella haemolytica were evaluated by measuring immunoglobulin production in blood for 9 days and in pulmonary lavage fluid for 7 days after intrapulmonary inoculation. Clinical signs, pulmonary lesions, pulmonary and systemic inflammatory response, and amount of antigen in lavage fluid were used to evaluate the response of calves to challenge with P haemolytica. The pulmonary response consisted of production of IgG, IgE, and IgM antibodies to P haemolytica antigens and a 17- to 68-fold increase of cells in lavage fluid 8 hours after inoculation, with a gradual decrease toward normal. Antibodies of the IgM isotype to P haemolytica were demonstrated as early as 8 hours through 7 days after inoculation in 3 of 3 calves. Of the anti-P haemolytica isotypes, IgM was found in the highest concentration. In all of the inoculated calves, IgE was found 1 to 2 days after inoculation, and IgG was found in 2 of 3 inoculated calves from day 1 through 7 after inoculation. Detection of IgG correlated with smaller pulmonary lesions. Immunoglobulin A was not detected in lavage fluid. Serum was evaluated for IgG and IgM antibody response to P haemolytica. Specific IgM was detectable 5 days after inoculation, and IgG was detectable 7 days after inoculation. Pasteurella haemolytica antigens were not detected in serum or plasma. A transient increase in neutrophil count was found 8 hours after inoculation, with return to baseline values by 24 hours after inoculation.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of calf age and Salmonella bacterin type on ability to produce immunoglobulins directed against Salmonella whole cells or lipopolysaccharide.

A commercially available Salmonella bacterin was administered to Holstein calves starting at 1 to 19 weeks of age. Serum samples were obtained before administering bacterin and at 2-week intervals thereafter. An ELISA with Salmonella dublin lipopolysaccharide (LPS) or S dublin whole cells as antigen, was used to measure specific IgG and IgM responses. Antibody responses to LPS were not detected from calves < 12 weeks old inoculated with killed bacterin. Immunoglobulin responses to whole-cell antigen were detected from all age groups of calves inoculated with the same killed Salmonella bacterin. Calves < 11 weeks old are able to produce immunoglobulins to some whole-cell antigens, but are unable to produce anti-LPS immunoglobulins when inoculated with killed Salmonella bacterin. This age-related response to killed Salmonella antigens may account, in part, for increased susceptibility to salmonellosis in calves < 12 weeks old. In comparison to the response for killed antigen, 8 calves given modified-live aromatic-dependent S dublin bacterin at 1 to 3 weeks of age had detectable anti-LPS immunoglobulins after immunization, although the response was not as rapid and was of a lesser magnitude than that of older calves given killed Salmonella bacterin.


Monophosphoryl lipid A-induced immune enhancement of Brucella abortus salt-extractable protein and lipopolysaccharide vaccines in BALB/c mice.

A study was conducted to determine the effect of monophosphoryl lipid A (MPL) and trehalose dimycolate (TDM) as adjuvants on the protective responses in BALB/c mice vaccinated with Brucella abortus salt-extractable protein (BCSP) or proteinase-K-treated B abortus lipopolysaccharide (PKLPS). Mice were vaccinated with different doses of BCSP or PKLPS given alone or in combination with MPL or TDM. Mice were challenge-exposed 4 weeks later with virulent B abortus strain 2308. Two weeks after challenge exposure, the number of B abortus colony-forming units (CFU) per spleen, spleen weights, and spleen cell interleukin 1 production were measured. Serum IgG and IgM concentrations specific for vaccinal immunogens were measured before and after challenge exposure with B abortus. Spleen weights and mean B abortus CFU per vaccine group were significantly lower in BCSP- and PKLPS-vaccinated mice, compared with those of nonvaccinated control mice. Monophosphoryl lipid A enhanced the suppression of splenic infection when given with the BCSP vaccine, but not when given with the PKLPS vaccine. Trehalose dimycolate had no effect on mean CFU when given with BCSP, but incorporation of TDM resulted in a significant increase in mean CFU when given with PKLPS. Spleen weights in BCSP- or PKLPS-vaccinated mice were not different when these vaccines were combined with MPL or TDM. Because of the wide variation in the results, we could not conclude that vaccination with BCSP or PKLPS alone, or in combination with MPL altered spleen cell interleukin-1 production in B abortus-infected mice.(ABSTRACT TRUNCATED AT 250 WORDS)


Evaluation of clinical signs of disease, bronchoalveolar and tracheal wash analysis, and arterial blood gas tensions in 13 horses with chronic obstructive pulmonary disease treated with prednisone, methyl sulfonmethane, and clenbuterol hydrochloride.

We evaluated the efficacy of 3 treatments for chronic obstructive pulmonary disease in horses: prednisone (400 mg/horse, PO, daily; n = 7), methyl sulfonmethane (10 g/horse, PO, q 12 h; n = 6), and clenbuterol hydrochloride (0.4 mg/horse, PO, q 12 h; n = 7). A fourth group acted as controls (n = 6) and was not treated. The treatment period lasted 10 days. Each horse was a member of 2 different groups for 10 days, separated by an 18-day interval of no treatment. All horses were housed together in an outdoor pen without bedding. Horses were fed alfalfa/grass hay mix ad libitum from a large feeder. The same batch of hay was fed throughout the study. Multiple physical and laboratory variables were monitored prior to, during, and at the end of each 10-day trial period. Changes in lung sounds, respiratory effort, degree of anal movement, nasal discharge, temperature, respiratory rate, or heart rate were not significant. Changes in arterial blood gas tensions, tracheal wash or bronchoalveolar lavage cytologic findings, or phagocyte function were not significant. All horses were tachypneic and most were tachycardic. The median value for PaO2 was below normal for all horses. All tracheal wash and most bronchoalveolar lavage cytologic findings represented a suppurative response. Negative linear correlation was observed between PaO2 and degree of respiratory effort in these horses (eg, as PaO2 decreased, the degree of respiratory effort increased).


Ultrastructural mucosal injury after experimental ischemia of the ascending colon in horses.

The ultrastructural injury that develops sequentially in the ascending colon during experimentally induced ischemia was examined in 6 halothane-anesthetized horses. Colonic ischemia was created by 2 types of vascular occlusion 24 cm proximal and distal to the pelvic flexure. In all horses, transmural vascular compression was created. The colonic venous circulation was obstructed in 3 horses, whereas in the other 3 horses, arterial and venous circulation was obstructed. Two additional horses were anesthetized as controls for determination of any morphologic alterations associated with the experimental protocol. Full-thickness colonic biopsy specimens were obtained from the antimesenteric border of the pelvic flexure at 0, 0.25, 0.5, 1, 1.5, 1.75, 2, 2.25, 2.5, 3, 3.5, 4, 4.5, and 5 hours during occlusion, and were studied by light and transmission electron microscopy. Morphologic alterations did not develop in the colon of control horses. Mucosal congestion was observed by light microscopy in the colon of horses with experimentally induced ischemia, but congestion developed early in those with obstructed colonic venous circulation, compared with those having arterial and venous obstruction. Inter- and intracellular vacuolation and loss of staining initially resulted in groups of 3 to 5 superficial luminal epithelial cells. Alterations in the glandular epithelium lagged behind those in the superficial epithelium, but were observed in both groups by 2 hours of obstruction. These changes progressed to 100% sloughing of all epithelium by 4.5 to 5 hours. The initial cellular alterations, which were observed by transmission electron microscopy, developed at 0.25 hour in horses with colonic venous obstruction and was characterized by inter- and intracellular edema.(ABSTRACT TRUNCATED AT 250 WORDS)


Bromodeoxyuridine labeling and DNA content of pulmonary arterial medial cells from hypoxia-exposed and nonexposed healthy calves.

Vascular medial thickening is a prominent finding in people and animals with refractory neonatal pulmonary hypertension. Smooth muscle cells are capable of 2 distinct growth responses in vivo: hypertrophy or hyperplasia. Hypertrophic smooth muscle cells may undergo DNA synthesis without cell division, leading to a polyploid state. To better understand the nature of smooth muscle cell growth in healthy and pulmonary hypertensive neonatal calves, we measured incorporation of the thymidine analog bromodeoxyuridine (BrdUrd) and total DNA content in medial cells from control (pulmonary arterial pressure = 32 +/- 2 mm of Hg) and hypobaric hypoxia-exposed (pulmonary arterial pressure = 120 +/- 7 mm of Hg) calves. Labeling of medial cells with BrdUrd measured by flow cytometry was increased (P < 0.02) in pulmonary arteries of hypoxia-exposed calves (n = 5), compared with control calves (n = 5). Immunohistochemical localization of BrdUrd indicated that BrdUrd labeling of large elastic pulmonary arteries from hypoxia-exposed calves was increased almost exclusively in the outer half of the medial wall. Increased BrdUrd labeling of muscular pulmonary arteries from hypoxia exposed calves was observed in the arterial media and adventitia, and tended to exit in clusters. Analysis of DNA content by flow cytometry indicated a decrease (P < 0.05) in percentage of tetraploid medial cells in pulmonary arteries from hypoxia-exposed calves, compared with control calves. Bivariate analysis for BrdUrd labeling and DNA content of cells from the pulmonary arteries of hypoxia-exposed calves indicated a subpopulation of diploid cells with positive BrdUrd labeling, suggestive of DNA synthesis and subsequent cell division.(ABSTRACT TRUNCATED AT 250 WORDS)


Retrospective evaluation of factors associated with the risk of seropositivity to Ehrlichia risticii in horses in New York State.

A retrospective study was designed to determine the distribution of equine monocytic ehrlichiosis among the equine population in New York state, and to identify factors associated with risk of disease. Serum samples submitted to the diagnostic laboratory of the university during the period from January 1985 through December 1986 were examined for antibodies to Ehrlichia risticii, using the indirect fluorescent antibody technique. Factors evaluated included geographic origin and date of submission of the sample, and age, breed, and sex of the horse. Logistic regression analysis was used to identify which factors were significantly associated with the risk of seropositivity to E risticii, while simultaneously controlling for other factors. Of the 2,579 tested samples, 1,950 (76%) had positive results. Factors significantly associated with risk of seropositivity to E risticii were: breed of the horse (Thoroughbreds were 3 times more likely to have been exposed to E risticii, compared with non-Standardbred, non-Thoroughbred breeds); sex (female horses were 2.7 times more likely to have been exposed, compared with male horses); age of the horse (the risk of being exposed to E risticii increased with age, peaked at around 12 years, and decreased thereafter); and month of submission (horses tested during November and December had the highest odds of being seropositive [odds ratio = 2.1], and horses tested during March through April were least likely to be seropositive [odds ratio = 0.5], compared with horses tested during January and February).


Small intestinal bacterial overgrowth and enhanced intestinal permeability in healthy beagles.

The small intestine of healthy adult Beagles was examined to determine whether subclinical abnormalities might exist that would be relevant to the use of Beagles in pharmacologic studies. Duodenal juice was obtained for qualitative and quantitative bacteriologic examinations; jejunal mucosa was taken for morphologic and biochemical investigation, and intestinal permeability was assessed by quantification of 24-hour urinary excretion of 51Cr-labeled EDTA after its oral administration. Comparisons were made with findings in healthy adult dogs of other breeds that served as controls. Small intestinal bacterial overgrowth was found in 14 of the 21 Beagles examined, and represented a mixed flora that included obligate anaerobic bacteria in 8 dogs and exclusively aerobic bacteria in 6 dogs. Intestinal permeability (percentage urinary recovery of 51Cr-labeled EDTA; mean +/- SEM) was considerably higher (P < 0.01) in Beagles with anaerobic overgrowth (37.6 +/- 3.2%) or aerobic overgrowth (30.5 +/- 4.8%), compared with Beagles with no overgrowth (17.3 +/- 1.6%) and with controls (11.1 +/- 1.0%). In Beagles, significant (r = 0.54, P = 0.03) correlation was observed between 24-hour urinary recovery of 51Cr-labeled EDTA and bacterial numbers in duodenal juice. Morphologic changes in jejunal mucosa were minimal, and specific activities of brush border enzymes were not significantly decreased, apart from aminopeptidase N, but activities of lysosomal and endoplasmic reticular marker enzymes were higher in the 3 groups of Beagles with anaerobic, aerobic, or no overgrowth, compared with controls.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of protein source in liquid formula diets on food intake, physiologic values, and growth of equine neonates.

The effects of 2 liquid formula diets differing in protein source were evaluated in orphan foals. The response of 7 foals fed a diet containing casein as the protein source, and 6 foals fed a diet containing a combination of whey and casein, was compared with the response in a reference group of 8 mare-raised foals. Orphaned foals were fed 150 kcal/kg of body weight/d, divided into 6 equal feedings of 25 kcal/kg. Formula intake was comparable among the experimental groups, and foals fed the liquid formula diet grew as well as mare-raised foals. There was no difference among groups in mean daily body weight gain, wither height, heart girth, body temperature, pulse, respiration rate, capillary refill time, or skin tenting. Insulin and blood glucose concentrations increased in both groups of foals fed formula diets, returning to prefeeding values within 4 hours. Differences among groups were found for serum alkaline phosphatase, alanine transaminase, cholesterol, creatinine, and glucose values; all other serum chemical values were comparable among groups. Plasma amino acid determinations revealed that arginine and ornithine were significantly lower in foals in both experimental groups than in reference foals, suggesting that arginine may have been the limiting amino acid in these diets. Diarrhea developed in foals in all treatment groups, but in most cases was self-limiting. These results suggest that the protein source of liquid formula diets may be less important in foals than in infants.


Malabsorption of vitamin A in preruminating calves infected with Cryptosporidium parvum.

Serum retinol, retinyl palmitate, and total vitamin A concentrations, and jejunoileal morphology were examined in neonatal calves infected with Cryptosporidium parvum. Group-1 calves served as noninfected controls and, after an adjustment period, were given 50 ml of saline solution i.v. every 12 hours for 6 days. Group-2 calves were inoculated with 10(7) C parvum oocysts and, after the onset of diarrhea, were given 50 ml of saline solution i.v. every 12 hours for 6 days. Group-3 calves were inoculated with 10(7) C parvum oocysts and, after the onset of diarrhea, were treated with difluoromethylornithine (DFMO, 200 mg/kg of body weight i.v., q 12 h) for 6 days. Group-4 calves were naturally infected with C parvum. Jejunoileal biopsy specimens were excised from calves of groups 1-3 at 3 and again at 15 to 16 days of age. During the course of diarrhea and 3 days after saline or DFMO administration, water-miscible retinyl palmitate was administered orally (2,750 micrograms/kg) to each calf in each group. Cryptosporidium parvum infection was associated with significant (P < or = 0.05) reduction in postadministration serum retinol, retinyl palmitate, and total vitamin A concentrations in calves of groups 2, 3, and 4. Cryptosporidium parvum infection caused significant (P < or = 0.05) reduction in villus height. Decreased villus height, villus blunting and fusion, and attenuation of the intestinal mucosa were associated with reduced absorption of vitamin A, as indicated by lower peak postadministration retinyl palmitate concentration in C parvum-infected calves. Intravenous administration of DFMO to group-3 calves did not improve retinol absorption.(ABSTRACT TRUNCATED AT 250 WORDS)