Development of reference intervals for serum biochemistry and haematology of juvenile green sea turtles (Chelonia mydas) in a Thai rehabilitation centre.

No reference intervals for serum biochemistry and haematology of sea turtles in Thailand exists to assist veterinarians who are responsible for sea turtle health management and treatment. This study determined serum biochemistry and basic haematology of healthy juvenile green sea turtles (n = 92) in captivity in Thailand following the American Society for Veterinary Clinical Pathology (ASVCP), Quality Assurance and Laboratory Standards Committee (QALS) guidelines for the determination of reference intervals in veterinary species. Biochemistry tests, including blood urea nitrogen, creatinine, uric acid, alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase were analysed using an IDEXX VetTest Chemistry Analyzer. Haematology parameters were measured manually using a microhaematocrit for packed cell volume (PCV), Neubauer counting chamber for red blood cell count and cyanmethemoglobin method for haemoglobin concentration. mean corpuscular volume and mean corpuscular haemoglobin concentration were calculated using the PCV, red blood cell count and haemoglobin. Turtles in this study were found to have higher mean values for PCV (28.70%), haemoglobin (92.13 g/L), mean corpuscular haemoglobin concentration (327.03 g/L), uric acid (247.15 µmol/L), alanine aminotransferase (16.53 IU/L), aspartate aminotransferase (209.44 IU/L), and alkaline phosphatase (245.08 IU/L) compared to sea turtles in Brazil. The reference intervals established using high numbers of healthy turtles in this study will assist veterinarians with diagnostic and treatment decisions when evaluating laboratory results for juvenile green sea turtles.

Comparison of ionised calcium measured using a portable analyser to a reference method in healthy dogs.

OBJECTIVES: To compare the ionised calcium measured on a portable analyser (iSTAT, Abbott) to a reference method. MATERIALS AND METHODS: Blood samples from 39 apparently healthy dogs were analysed in duplicate using a portable analyser and a reference method (Radiometer ABL800 FLEX). Bland-Altman plots and Passing-Bablok regression were used to assess constant and proportional bias between the two instruments. A within-assay percentage coefficient of variation and total error (TE) was calculated for both analysers. The reference interval was calculated for the portable analyser using the robust method with confidence interval bootstrapping. RESULTS: The Bland-Altman plot showed a -0.036 mmol/L difference between the two instruments (95% confidence limit -0.08 to 0.01 mmol/L; limits of agreement -0.07 to 0.006 mmol/L). Neither the Bland-Altman plot nor the Passing-Bablock regression (slope -0.03; 95% confidence interval -0.08 to 0.19 and intercept 1; 95% confidence interval 0.83 to 1.2) showed significant proportional bias. The coefficient of variation for the portable analyser was 1.08%, compared to 0.78% for the reference method with a total error of 3.5% for the portable analyser. The estimated population-based reference interval for ionised calcium using the portable analyser is 1.23 to 1.42 mmol/L. CLINICAL SIGNIFICANCE: For the healthy dogs in this study, compared to the reference method, the portable analyser showed no significant bias for measurement of ionised calcium. Further studies including hyper and hypocalcaemic dogs are required to determine clinical impact of the use of this analyser.

Urinary bladder botryoid rhabdomyosarcoma with widespread metastases in an 8-month-old Labrador cross dog.

An 8-month-old crossbred Labrador retriever was presented with a history and clinical signs suggestive of lower urinary tract obstruction. Laboratory results revealed azotaemia and hyperphosphataemia. Ultrasonographic evaluation of the urinary tract showed a mass at the bladder trigone, hydronephrosis, hyrodureter, and suspected metastases to lymph nodes and the liver. Pulmonary metastasis was identified on thoracic radiographs. A post mortem confirmed metastases to the liver, lungs and regional lymph nodes, as well as to the mesenteric lymph nodes, mediastinum, heart, subcutaneous tissue and several muscle groups. A histopathological diagnosis of metastatic botryoid rhabdomyosarcoma (sarcoma botryoides) was made. A review of the literature shows that, although the bladder trigone is a well documented location for this tumour, this case was unique with its widespread metastases to previously undocumented organs. The incidence, embryology, ultrasonographic appearance and treatment of this tumour are discussed.

Comparison of mid-infrared and Raman spectroscopy in the quantitative analysis of serum.

Mid-infrared or Raman spectroscopy together with multivariate data analysis provides a novel approach to clinical laboratory analysis, offering benefits due to its reagent-free nature, the speed of the analysis and the possibility of obtaining a variety of information from one single measurement. We compared mid-infrared and Raman spectra of the sera obtained from 247 blood donors. Partial least squares analysis of the vibrational spectra allowed for the quantification of total protein, cholesterol, high and low density lipoproteins, triglycerides, glucose, urea and uric acid. Glucose (mean concentration: 154 mg/dl) is frequently used as a benchmark for spectroscopic analysis and we achieved a root mean square error of prediction of 14.7 and 17.1 mg/dl for mid-infrared and Raman spectroscopy, respectively. Using the same sample set, comparable sample throughput, and identical mathematical quantification procedures Raman and mid-infrared spectroscopy of serum deliver similar accuracies for the quantification of the analytes under investigation. In our experiments vibrational spectroscopy-based quantification appears to be limited to accuracies in the 0.1 mmol/l range.

Biochemical and genetic studies of the initiation of human rhinovirus 2 RNA replication: identification of a cis-replicating element in the coding sequence of 2A(pro).

We have previously shown that the RNA polymerase 3D(pol) of human rhinovirus 2 (HRV2) catalyzes the covalent linkage of UMP to the terminal protein (VPg) using poly(A) as a template (K. Gerber, E. Wimmer, and A. V. Paul, J. Virol. 75:10969-10978, 2001). The products of this in vitro reaction are VPgpU, VPgpUpU, and VPg-poly(U), the 5' end of minus-strand RNA. In the present study we used an assay system developed for poliovirus 3D(pol) (A. V. Paul, E. Rieder, D. W. Kim, J. H. van Boom, and E. Wimmer, J. Virol. 74: 10359-10370, 2000) to search for a viral sequence or structure in HRV2 RNA that would provide specificity to this reaction. We now show that a small hairpin in HRV2 RNA [cre(2A)], located in the coding sequence of 2A(pro), serves as the primary template for HRV2 3D(pol) in the uridylylation of HRV2 VPg, yielding VPgpU and VPgpUpU. The in vitro reaction is strongly stimulated by the addition of purified HRV2 3CD(pro). Our analyses suggest that HRV2 3D(pol) uses a "slide-back" mechanism during synthesis of the VPg-linked precursors. The corresponding cis- replicating RNA elements in the 2C(ATPase) coding region of poliovirus type 1 Mahoney (I. Goodfellow, Y. Chaudhry, A. Richardson, J. Meredith, J. W. Almond, W. Barclay, and D. J. Evans, J. Virol. 74:4590-4600, 2000) and VP1 of HRV14 (K. L. McKnight and S. M. Lemon, RNA 4:1569-1584, 1998) can be functionally exchanged in the assay with cre(2A) of HRV2. Mutations of either the first or the second A in the conserved A(1)A(2)A(3)CA sequence in the loop of HRV2 cre(2A) abolished both viral growth and the RNA's ability to serve as a template in the in vitro VPg uridylylation reaction.

Biochemical and genetic studies of the initiation of human rhinovirus 2 RNA replication: purification and enzymatic analysis of the RNA-dependent RNA polymerase 3D(pol).

The replication of human rhinovirus 2 (HRV2), a positive-stranded RNA virus belonging to the Picornaviridae, requires a virus-encoded RNA polymerase. We have expressed in Escherichia coli and purified both a glutathione S-transferase fusion polypeptide and an untagged form of the HRV2 RNA polymerase 3D(pol). Using in vitro assay systems previously described for poliovirus RNA polymerase 3D(pol) (J. B. Flanegan and D. Baltimore, Proc. Natl. Acad. Sci. USA 74:3677-3680, 1977; A. V. Paul, J. H. van Boom, D. Filippov, and E. Wimmer, Nature 393:280-284, 1998), we have analyzed the biochemical properties of the two different enzyme preparations. HRV2 3D(pol) is both template and primer dependent, and it catalyzes two types of synthetic reactions in the presence of UTP, Mn(2+), and a poly(A) template. The first consists of an elongation reaction of an oligo(dT)(15) primer into poly(U). The second is a protein-priming reaction in which the enzyme covalently links UMP to the hydroxyl group of tyrosine in the terminal protein VPg, yielding VPgpU. This precursor is elongated first into VPgpUpU and then into VPg-linked poly(U), which is identical to the 5' end of picornavirus minus strands. The two forms of the enzyme are about equally active both in the oligonucleotide elongation and in the VPg-primed reaction. Various synthetic mutant VPgs were tested as substrates in the VPg uridylylation reaction.

Myocardial tissue characterization in heart failure by real-time integrated backscatter.

OBJECTIVE: Differentiation between normal and abnormal physical state of the myocardium, not possible with conventional echocardiography, so far could be done with integrated backscatter (IBS) as a research tool only. METHODS: This study investigates myocardial texture analysis with new commercially available real time IBS in 12 normal individuals and in 18 patients with severe left ventricular dysfunction due to coronary artery disease (CAD) in 8 and dilated cardiomyopathy (DCM) in 10 patients. Analysis of IBS amplitude and cyclic variation (dB) in the parasternal long and short axis view of the septum and the posterior wall were measured and corrected with IBS curve of the blood to get absolute values. RESULTS: Compared to normal individuals patients with left ventricular dysfunction had a reduced myocardial cyclic variation (P<0.0001), which correlated to regional systolic wall thickening (r=0.64, P=0.001) and global shortening fraction (r=0.62, P<0.01). Although systolic wall thickening in the posterior wall was lower in CAD patients (% thickening, 11.9+/-10 vs. 21.9+/-8, P=0.004), absolute cyclic variation was reduced in both, CAD and DCM patients in the same order of magnitude. However, the higher maximal IBS amplitude in the posterior wall observed in CAD when compared to DCM patients (13.2+/-4.4 vs. 9.2+/-2.4 dB; P=0.002) indicate fibrosis or scar. The dissociation between cyclic variation and systolic wall thickening could implicate hybernating myocardium. CONCLUSION: Real-time IBS has progressed from research to routine as a tool to obtain additional and valuable information to conventional echocardiography in daily practice.

A serotonin-1A receptor agonist and an N-methyl-D-aspartate receptor antagonist oppose each others effects in a genetic rat epilepsy model.

The WAG/RIJ rats exhibit spontaneously occurring spike-wave discharges (SWD) accompanied by behavioural phenomena, with characteristics similar to the human absence type epilepsy. To study the mechanisms involved in this type of epileptiform activity we investigated the effects of the serotonin-1A (5-HT1A) receptor agonist 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT) and the N-methyl-D-aspartate (NMDA) receptor antagonist (+)-5-methyl-10,11-dihydro-5H-dibenzo [a,d]cyclohepten-5,10-imine maleate (MK-801). Intracerebroventricular (i.c.v.) injection of 8-OH-DPAT caused marked, dose dependent increase, MK-801 a decrease in the cumulative duration and number of spike-wave discharges. Pretreatment with MK-801 (10 microg/rat i.c.v.) abolished the increase caused by 8-OH-DPAT (20 microg/rat i.c.v.), but the decrease in SWD to MK-801 was counterbalanced by 8-OH-DPAT. These data provide evidence for an interaction of glutamatergic and serotonergic mechanisms in the triggering and maintenance of epileptic activity in this genetic model of absence epilepsy.

The 5-HT1A agonist 8-OH-DPAT increases the number of spike-wave discharges in a genetic rat model of absence epilepsy.

The effects of the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT) on the epileptiform activity has been investigated in adult WAG/RIJ rats. Either intraperitoneal (0.1-0.5 mg/kg) or intracerebroventricular (2-20 microg/rat) administration of 8-OH-DPAT caused marked, dose-dependent increases in the number and mean cumulative duration of spike-wave discharges. These effects were attenuated by NAN-190, a 5-HT1A receptor antagonist. These data indicate that serotonergic system regulates the epileptiform activity in this genetic model of human absence epilepsy.

Experimental studies on the antitussive properties of the new xanthine derivative 1H-purine-2,6-dione, 3,7-dihydro-3-methyl-7[(5-methyl-1,2,4-oxadiazol-3-yl) methyl]. 3rd communication: examinations on opioid mechanisms and physical drug dependence.

CH-13584 (formerly: KHL-8425, 1H-purine-2,6-dione, 3,7-dihydro-3-methyl-7[(5-methyl-1,2,4-oxadiazol-3-yl)methyl], CAS 115779-20-9) showed antitussive effect on the citric acid spray-induced cough model. The antitussive effect of p.o. CH-13584 was antagonised by i.m. or intracerebroventricular (i.c.v.) naloxone, i.m. nor-binaltorphimine or s.c. beta-funaltrexamine. Intracerebroventricular administration of CH-13584 induced long-lasting antitussive effect which was antagonised by coadministration of i.c.v. naloxone. CH-13584 did not bind to opioid mu, delta, kappa receptor in vitro or inhibit the [3H]diprenorphine binding in vivo. Two-week treatment with CH-13584 up to the dose of 100 mg/kg p.o. did not produce autonomic and behavioural signs of withdrawal induced either by drug withdrawal or by naloxone injection, while morphine and codeine induced characteristic opioid-type physical dependence in rats.

High-resolution monochromatic x-ray imaging system based on spherically bent crystals.

We have developed an improved x-ray imaging system based on spherically curved crystals. It is designed and used for diagnostics of targets ablatively accelerated by the Nike KrF laser. A spherically curved quartz crystal (d = .?, R = mm) has been used to produce monochromatic backlit images with the He-like Si resonance line (1865 eV) as the source of radiation. The spatial resolution of the x-ray optical system is 1.7 mum in selected places and 2-3 mum over a larger area. Time-resolved backlit monochromatic images of polystyrene planar targets driven by the Nike facility have been obtained with a spatial resolution of 2.5 mum in selected places and 5 mum over the focal spot of the Nike laser.

2-Phenyl-4-(aminomethyl)imidazoles as potential antipsychotic agents. Synthesis and dopamine D2 receptor binding.

A series of 2-phenyl-4-(aminomethyl)imidazoles were designed as conformationally restricted analogs of the dopamine D2 selective benzamide antipsychotics. The title compounds were synthesized and tested for blockade of [3H]YM-09151 binding in cloned African green monkey dopamine D2 receptor preparations. The binding affinity data thus obtained were compared against that of the benzamides and a previously described series of 2-phenyl-5-(aminomethyl)-pyrroles.

Association of the Red Ring Nematode and Other Nematode Species with the Palm Weevil, Rhynchophorus palmarum.

The palm weevil, Rhynchophorus palmarum (L.), was collected in cocoons from red ring-diseased coconut palms (Cocos nucifera L.) in Trinidad and Tobago. Juveniles of five species of nematodes were extracted from the genitalia and macerated bodies of newly emerged adults of the palm weevil: Rhadinaphelenchus cocophilus (Cobb) Goodey (the red ring nematode), Teratorhabditis sp., Diplogasteritus sp., Mononchoides sp., and Bursaphelenchus sp. Over 90% of newly emerged weevil females and males were infested internally with red ring nematode juveniles, and over 47% of the weevils contained more than 1,000 red ring nematodes each. There was no significant correlation between weevil body length and the number of red ring nematodes carried internally by each weevil. Teratorhabditis sp. and Diplogasteritus sp. were extracted from over 50% of the palm weevils, and Monochoides sp. and Bursaphelenchus sp. were found in a small proportion of the weevils. Field-collected adult weevils were also internally and externally infested with a Rhabditis sp., which was not observed in or on weevils allowed to emerge from field-collected cocoons.

Teratorhabditis palmarum n. sp. (Nemata: Rhabditidae): An Associate of Rhynchophorus palmarum and R. cruentatus.

Teratorhabditis palmarum n. sp., an associate of the palm weevils Rhynchophorus palmarum and R. cruentatus is described. Teratorhabditis palmarum was isolated from newly emerged adults and cocoons of R. palmarum from red-ring diseased coconut palms, Cocos nucifera, in Trinidad and Ecuador, and from red-ring diseased oil palms, Elaeis guineensis, in Colombia. Teratorhabditis palmarum was also associated internally with newly emerged adults of R. cruentatus from mature transplanted cabbage palmettos, Sabal palmetto, in Florida. Dauer juveniles of T. palmarum infested the genital capsule and body cavity of newly emerged adult female and male palm weevils. Adult nematodes isolated from cocoons and dauer juveniles from newly emerged palm weevils were cuhurable on bacterial lawns on several solid media. Females of T. palmarum have a single anteriorly directed ovary; vulva at 93-96% of the body length; short, hemispherical spicate tail; three or four teeth in the metastom; cuticle with distinct transverse punctations that change abruptly at the level of the procorpus to indentations of alternating size and arrangement; and eggs with cuticular sculpturing. Males have a crenate, peloderan bursa with a 2 + 5 + 3 pattern of bursal rays (7 extend to the margin of bursa); spicules linear, completely fused at the distal tip and dorsally for 50% of the total spicule length.

Effect of antacid treatment on endogenous prostaglandin synthesis in human antral and duodenal mucosa.

Using 14C-labeled arachidonic acid as precursor for in vitro prostaglandin synthesis, the effect of an antacid containing Al (OH)3, Mg(OH)2 and CaCO3 on endogenous prostaglandin synthesis was investigated in antral and duodenal mucosa of healthy volunteers. After three weeks of treatment with a high-dose antacid, there was no detectable change in the total capacity of the mucosa for prostaglandin synthesis, but the prostaglandin profile was markedly altered. The relative amounts of PGE2 and PGF2 alpha synthesized by antral and duodenal mucosa increased at the expense of the prostaglandins A2/B2, thromboxane A2, and prostacyclin. In a short-term study, this change was not observed following a single antacid dose within 1 hr after application. It is concluded that long-term antacid treatment may alter the prostaglandin pattern formed by gastroduodenal mucosa and this may be related to its therapeutic effect.

Transient changes in cyclic AMP and in the enzymic activity of protein kinase and phosphorylase during the cardiac cycle in the canine myocardium and the effect of propranolol.

Oscillation of cyclic AMP and in the activity ratio of cyclic AMP-dependent protein kinase and of glycogen phosphorylase with the cardiac cycle were demonstrated in the canine heart in situ. For tissue sampling an ECG (R-wave)-triggered, automatically working push-freeze-drill apparatus was developed which allows intraventricular cryobiopsies from the left ventricular muscle of anaesthetized open-chest dogs. The nucleotide cyclic AMP oscillated with the cardiac cycle during normal working condition, the higher cyclic AMP level occuring during systole. Cyclic GMP was assayed to be without oscillatory changes during the contraction-relaxation cycle. The rise in the activity ratio of protein kinase was found to coincide with the maximum in the level of cyclic AMP. Propranolol pretreatment prevents the transient in the level of the nucleotide as well as in the activity ratio of the kinase indicating i) a causal relationship between these changes and ii) a neurohumoral, beat-to-beat regulation by catecholamines released from the sympathetic nerve endings within the heart. Contrary the activity ratio of phosphorylase retains its transient changes during the cardiac cycle in the presence of propranolol, indicating a Ca-mediated activation of phosphorlase kinase during the contraction process.

Alimentary tract and pancreas. Stimulation of mucosal prostaglandin synthesis in human stomach and duodenum by antacid treatment.

The effect of a low dose antacid treatment on mucosal prostaglandin metabolism was studied in 15 healthy volunteers. A daily dose of 46 mmol (= 138 mval) Al(OH)3 and 42 mmol (= 84 mval) Mg(OH)2 with a titrated in vitro neutralising capacity of 272 mval of H+ was given for three weeks. Total prostaglandin formation and the prostaglandin profile as well as the degradation of PGE2 were assayed by incubating homogenates of endoscopic biopsies from antral and duodenal mucosa with the precursor (14C)arachidonic acid. Total prostaglandin synthesis in antrum (623 (110) pmol/mg protein) and duodenum (432 (72) pmol/mg) was stimulated after three weeks administration of low dose antacids by 176% (p less than 0.05) and 154% (p less than 0.05), respectively. An untreated control group exhibited no significant changes. In contrast, the prostaglandin profile showed only a small increase of the prostacyclin metabolite 6-keto PGF1a (p less than 0.05) at the expense PGD2. PGE2 catabolism was unaffected. This enhanced activity of mucosal prostaglandin cyclooxygenase might represent one possible mechanism of action of a low dose antacid treatment.

[Prostaglandin synthesis in stomach and duodenal mucosa of the human: effect of aspirin with and without antacid]. / Die Prostaglandinsynthese in Magen- und Duodenalmukosa des Menschen: Einfluss von Aspirin mit und ohne Antazidum.

A decreased PG E2 content in gastric mucosa of humans receiving a longterm antirheumatic therapy has been reported to be partially reversed after a one week treatment with an antacid (Reimann et al., Fortschr Med 102 [1984], 25-26). Therefore the effect of an Aspirin treatment of healthy volunteers with or without an antacid on the prostaglandin synthesis in gastric or duodenal mucosa was investigated. 14C-labelled arachidonic acid was used as substrate during in vitro incubation of the mucosal homogenate for determination of the endogenous formation of prostaglandins. Aspirin suppressed the total prostaglandin synthesis to less than 10% of control after one week treatment. The addition of a high dose antacid did not influence this inhibition, and no significant effect on the prostaglandin profile was detectable. It is concluded that antacids do not influence the suppression of the endogenous prostaglandin synthesis by NOSAC's, however another effect, eg a prolonged stability of PG E2 in a less acidic environment is more likely.