Pharmocokinetic and pharmacodynamic evaluation of intravenous morphine in dogs

Morphine is considered the prototypical opiate analgesic. Despite the common use of morphine in dogs, ideal dosing strategies have not been formulated due to the difficulty in assessing its analgesic effects. The purpose of this study was to: 1) evaluate a noninvasive mechanical threshold device (von Frey device) to measure antinociceptive responses (pharmacodynamics) of opiates in dogs and 2) evaluate the pharmacokinetics (PK) and pharmacodynamics (PD) of intravenous (IV) morphine in dogs. Six healthy Beagle dogs were used. The von Frey threshold (vFT) response was evaluated hourly for 8 hours in each dog to examine the effect of repeated testing (controls). PK and PD (vFT) measurements were then made following a 1 mg kg^–1 IV bolus of morphine sulfate. A two way blinded crossover consisted of an 8 hour IV constant rate infusion of saline or morphine with hourly PD measurements. The individual CRI was based on individual PK data and adjusted every 2 hours to attain targeted plasma concentrations of morphine of 10, 20, 30, and 40 ng mL^–1. Blood samples were taken hourly in all phases, except the controls. No significant (p > 0.05) intraindividual changes in vFT occurred in the controls over 8 hours. The morphine bolus produced increased vFT at 1, 2, 3, and 4 hours post injection (p < 0.05). The E_MAX and EC₅₀ following the IV bolus were 213 ± 104% (increase from baseline) and 13.9 ± 5.8 ng mL^–1, respectively. The CRI produced increased vFT at plasma concentrations >30 ng mL^–1, when compared to saline controls (p < 0.05). Targeted plasma concentrations were inconsistent at higher infusion rates, suggesting the PK of morphine may change during CRI. The actual mean ± SD CRI plasma concentrations (ng ml^–1) were 10.8 ± 3.0, 22.7 ± 7.4, 32.4 ± 13.9, 35.7 ± 16.9. Morphine dosing protocols should be re‐evaluated, as sufficient analgesia may not be obtained from published dosages. Intravenous boluses may be more predictable than CRI.     

Estimation of microbial biomass potassium in paddy field soil

Abstract

Potassium (K) in microbial cells, microbial biomass K, in soil has been recently recognized as a K pool for plant growth. We determined soil microbial biomass K in paddy fields to reveal its importance as a K pool in paddy field soil for the first time. Microbial biomass K ranged from 5 to 21 mg K kg^?1 in the soil samples periodically collected from a paddy field and the value corresponded to 41% of the exchangeable K on average. Both microbial biomass K and exchangeable K increased conspicuously due to the long-term application of livestock manure compost or rice straw compost. Biomass K was higher than exchangeable K under K-deficient conditions in the long-term experimental plots without K application. The present study revealed that the microbial biomass contained considerable amounts of K in paddy field soil, indicating the need for evaluation of the microbial biomass K as a source and a stock of K in soil that has been overlooked.     

Activity of protease extracted from rice-rhizosphere soils under double cropping of rice and wheat

Abstract

Protease-active extracts were prepared from 2 rice-rhizosphere and non-rhizosphere soil samples 3 proteolytic isolates from the soil samples which were taken from water logged paddy fields (Gray Lowland Soils) under double cropping of rice and wheat that had been treated with chemical fertilizer (CF) or organic manure (OM). The activities were assayed and characterized using an artificial substrate, benzyloxycarbonyl-L-phenylalanyl-L-tyrosyl-L-leucine (ZFTL). The protease-active extracts from the rhizosphere and non-rhizosphere of the CF and OM soils (abbreviated as CFR, CFN, OMR, and OMN) showed an optimum pH between 7 and 8. The extracts hydrolyzed ZFTL to Tyr-Leu and Leu; generally, Tyr-Leu was the main reaction product. Activities of all the samples were inhibited by EDTA and p-chloromercuribenzoic acid (PCMB) but not by pepstatin and phenylmethanesulfonyl fluoride (PMSF). These results indicate that the main components of the ZFTL-hydrolases in CFR, CFN, 0MR, and OMN were essentially similar.

Optimum pH for the activity of the protease extracted from a Gram negative rod (MNB003)., two actinomycetous isolates (MNA014., MRA017) was about 8, 8, and 8.5, respectively. Extract from the culture of MNB003 hydrolyzed ZFTL to Tyr-Leu., Leu,., the protease activity was inhibited by EDTA., PCMB. Protease activities of the extracts from cultures of the two actinomycetes were inhibited by EDTA but not by PCMB. The extract from culture of MNA014 produced mainly Tyr-Leu from ZFTL while MRA017 produced Leu. Characteristics of the protease activity from cultures of MNB003., MNA014 were similar to those of the enzyme extracted from a paddy field soil under single cropping (Takeuchi., Hayano 1994: Soil Sci. Plant Nutr., 40, 691-695) and from an Andosol under tomato cultivation (Hayano et al. 1987: Biol. Fertil. Soils, 4, 179-183), respectively.     

Spirometra erinacei / S. erinaceieuropaei in a feral cat in Manawatu with chronic intermittent diarrhoea

CASE HISTORY: A feral cat captured in the Manawatu region of New Zealand was treated for worms and fleas, and kept confined in a metabolic cage. It showed good appetite and weight gain but had intermittent watery, yellow diarrhoea.

CLINICAL FINDINGS: Clinical examination under sedation was unremarkable and routine blood tests showed no significant abnormalities. The cat was negative for feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV). Different canned cat foods did not alter the course of the diarrhoea, and the cat was euthanised 6 months after capture. At necropsy, two sections of adult Spirometra tapeworms were found in the jejunum and typical Spirometra eggs were found in colonic contents. Molecular identification of the parasite was undertaken, using the cytochrome- c oxidase subunit-1 gene (cox1) sequence.

DIAGNOSIS: Chronic intermittent diarrhoea associated with Spirometra erinacei / S. erinaceieuropaei infection.

CLINICAL RELEVANCE: Spirometra has not been reported in New Zealand before but has been associated with gastrointestinal disease in cats in other parts of the world. It requires speciestargeted treatment to be eliminated effectively, and is zoonotic. Diagnosis could be diffi cult for clinicians who are not familiar with the parasite and its life cycle.     

Distal aortic aneurysm presumed to be secondary to an infected umbilical artery in a foal

Abstract

CASE HISTORY:?A 3-month-old female Warmblood foal was presented after displaying signs of colic with pyrexia for 5 days.

CLINICAL AND PATHOLOGICAL FINDINGS:?The foal continued to show signs of colic, frequently passed urine, and was pyrexic with an elevated white blood cell count. The umbilical stalk was thickened but there was no evidence of purulent material. Exploratory laparotomy revealed an enlarged left umbilical artery remnant tightly adhered to the bladder wall. The left umbilical artery continued to an aneurysm involving the distal aorta. The foal was subject to euthanasia and post-mortem examination confirmed a spherical aortic aneurysm, in the dorsal midline caudal to the kidneys that contained a large thrombus. Histopathological examination revealed inflammation and necrosis of the tunica intima and tunica media of the left umbilical artery with suppuration and bacterial colonies evident in the periarterial tissues.

DIAGNOSIS:?Infected aortic aneurysm presumably caused by an umbilical artery infection.

CLINICAL RELEVANCE:?A previously undetected umbilical infection appears to have resulted in an unusual delayed complication causing signs of colic in a foal. Veterinarians should be aware of this condition, and the possibility that it may be a cause of signs of colic in foals. Diagnosis based on ultrasonography should be possible, but may require sedation, visceral analgesia and careful examination.     

Expression of Apoptosis Regulatory Genes and Incidence of Apoptosis in Different Morphological Quality Groups of In Vitro‐produced Bovine Pre‐implantation Embryos

Apoptosis occurs during early development in both in vivo‐ and in vitro‐produced embryos, and is considered as one of the causes of embryonic loss. The objectives of this study were, therefore, investigating stage‐specific expression profiles of apoptosis regulatory genes in three quality groups of in vitro‐produced bovine pre‐implantation embryos; and analysing the relationship between cell number and DNA fragmentation with expressions of those genes. The relative abundance of mRNA of 9 pro‐ (Bax, caspase‐9, Bcl‐xs, P53, Caspase‐3 and Fas) and anti‐ (Bcl‐w and Mcl‐1) apoptotic genes was analysed. Differential cell staining and terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end labelling were performed to analyse the variation in cell numbers and detect apoptotic nuclei respectively. Expression of Bax and Caspase‐3 genes was significantly (p < 0.05) higher in poor quality pre‐implantation embryos as compared with that of morphologically good quality embryos of the same developmental stages. Moreover, Mcl‐1 expression was significantly higher in good quality immature oocytes than that in the poor quality group. Moreover, higher DNA fragmentation was evidenced in morphologically poor quality blastocysts. In conclusion, our study demonstrates that Bax, caspase‐3 and Mcl‐1 can be used as potential markers of embryo quality to evaluate in vitro‐produced bovine embryos. Further studies are required to investigate specific molecular signatures that can be used in evaluating in vivo‐derived embryos.     

Chemical constituents of the Vietnamese liverwort Porella densifolia

From the ether extract of Porella densifolia, the first liverwort collected and chemically investigated in Vietnam, a kaurane diterpenoid, ent-kauren-15-one (1) and two sesquiterpene norpinguisone (2) and norpinguisone methyl ester (3) have been purified. Their structures were elucidated by spectroscopic analysis. In addition, an X-ray crystal structure of norpinguisone was obtained, allowing to determine its stereochemistry. Furthermore, the inhibitory activity of nitric oxide (NO) production in RAW 264.7 cells stimulated by lipopolysaccharide (LPS) of compounds 1–3 was examined with their IC₅₀ values of 69.4, 45.5 and 1.68 μM, respectively.     

Differential expression of heat-shock proteins in F2 offspring from F1 hybrids produced between thermally selected and normal rainbow trout strains

Rainbow trout Oncorhynchus mykiss is a cold-water aquaculture species, and a thermally selected strain has been developed by multigenerational high-temperature breeding in Japan. We examined the expression of heat-shock proteins as candidates responsible for thermotolerance in rainbow trout using F2 offspring from F1 hybrids produced between thermally selected and normal strains. From F2 offspring, two groups were selected for western blot analysis, namely, low- and high-thermotolerance groups (times to loss of equilibrium were <30 and ??60?min, respectively). We demonstrated that the expression levels of Hsp70, Hsp60, and Hsp40 in tail fin tissues were significantly higher in the individuals with high thermotolerance than in those with low thermotolerance under non-heat-shock conditions. In particular, Hsp70 was expressed only in the individuals with high thermotolerance. These results suggest that Hsp70 is a major protein responsible for conferring thermotolerance in rainbow trout.