Cyclooxygenase ‐2 Expression in Mammary Tumors in Dogs and its Correlation to Histologic and Biologic Behavior

Introduction:  Cyclooxygenase‐2 (Cox‐2) is the inducible form and the rate‐limiting enzyme, for conversion of arachidonic acid to prostaglandins. Cox‐2 overexpression, common in carcinomas, is associated with increased growth rate, resistance to apoptosis, angiogenesis, and overall, both local and distant aggressive behavior. Cox‐2 overexpression has been detected in human and canine mammary tumors (MTs). Histopathology of canine MT is not always predictive of biologic behavior, and anecdotally, only 50% of the malignant MTs are expected to metastasize. We hypothesize that Cox‐2 expression correlates with aggressive behavior.
Methods:  This retrospective study evaluated 48 bitches, presented for excision of MT between 2000 and 2003 at FMVZ de Botucatu‐UNESP, Brasil. Follow‐up varied from 18 months to 24 months and included physical examination and thoracic radiographs. Histopathologic examination was performed in all tumors, as well as in metastatic lesions when detected in the follow‐up period. Immunohistochemistry was used to detect expression of Cox‐2 in paraffin blocks (Rabbit polyclonal anti‐PGHS‐2. Oxford Biomedical). 10 adenomas, 10 carcinomas, 10 benign mixed tumors, 10 malignant mixed tumors and 8 cases of primary carcinomas and their metastatic lesions.
Results:  Expression of Cox‐2 varied among groups. Adenomas (32.1%), mixed benign tumors (38%), carcinomas (60.3%), malignant mixed tumors (65.8%), and metastatic carcinomas (81.25%) and their metastatic lesions (84.35%). Statistically significant differences (p < 0.05) were observed between the benign and malignant counterparts and between carcinomas and metastatic carcinomas.
Conclusions:  Cox‐2 expression correlates with both histologic and biologic behavior in mammary carcinomas, and may serve as a predictor of metastatic potential.     

Effect of luteinising-hormone in vivo on ovarian cell subpopulations of newly-hatched chicks

1. We analysed the number and size of different ovarian cell subpopulations of newly-hatched chicks by ovarian cell suspension count and morphometric/stereological methods as well as delta5-3beta-hydroxysteroid dehydrogenase (delta5-3beta HSD) activity in these cells treated in vivo with LH during embryonic development. 2. Fertile White Leghorn eggs received 1 microg LH applied to the chorioallantoic membrane on days 13, 15, and 17 of incubation. All animals were killed within 24 h after hatching, the left ovary was dissected and processed. 3. The results indicate that the number of germ, pregranulosa, interstitial and undifferentiated cells was not affected by LH treatment. However, we observed an increase in the size of individual interstitial cells of the ovarian medulla. In these cells, delta5-3beta HSD activity was increased in response to LH. 4. These findings suggest that LH does not exert a proliferative effect on the cells of the prefollicular ovary of the chick and that interstitial cells can be target cells for LH, increasing their steroidogenic activity due to LH treatment.     

Ciliates in a waste stabilization pond system in Mexico

A protozoological survey was performed on a waste-stabilization pond to isolate, identify, and determine the incidence of ciliates present in these waters. The correlation between the species found and the physicochemical parameters was investigated. One hundred and eighty samples were analyzed from February 1983 to February 1984. Free-living ciliates isolated include:Carchesium polypinum Linnaeus,Vorticella microstoma Ehrenberg,Vorticella convallaria Linnaeus,Didinium balbiani Fabré-Domergue,Amphileptus claparedei Stein,Spathidium spathula Müller,Litonotus fasciola Ehrenberg,Paramecium caudatum Müller,Paramecium aurelia Ehrenberg,Tetrahymena vorax Kidder,Tetrahymena pyriformis Ehrenberg,Cyclidium glaucoma Müller,Aspidisca costata Ehrenberg, andStylonychia putrina Stokes. The performance of the pond was predominantly anaerobic. The temperature, total alkalinity, and nitrate concentration were the most variable parameters in the system studied.     

Detection of airborne inoculum of Leptosphaeria maculans and Pyrenopeziza brassicae in oilseed rape crops by polymerase chain reaction (PCR) assays

The potential use of DNA-based methods for detecting airborne inoculum of Leptosphaeria maculans and Pyrenopeziza brassicae , both damaging pathogens of oilseed rape, was investigated. A method for purifying DNA from spores collected using Hirst-type spore samplers and detecting it using polymerase chain reaction (PCR) assays is described. For both pathogens, the sensitivities of the DNA assays were similar for spore-trap samples and pure spore suspensions. As few as 10 spores of L. maculans or P. brassicae could be detected by PCR and spores of both species could be detected against a background of spores of six other species. The method successfully detected spores of P. brassicae collected using spore traps in oilseed rape crops that were infected with P. brassicae. Leptosphaeria maculans spores were detected using spore traps on open ground close to L. maculans -infected oilseed rape stems. The potential use of PCR detection of airborne inoculum in forecasting the diseases caused by these pathogens is discussed.     

Toxic effect of exposure to carbon monoxide onTribolium castaneum adults and pupae

Tribolium castaneum adults and pupae were exposed to carbon monoxide (CO) at concentrations of 20%, 30% and 40% in air, for 24, 48, 72, 96 and 120 h. The toxic effect on the treated adults was insignificant except for the longest exposure at the highest CO concentration, at which mortality values reached 11.3%. Adult emergence from the treated pupae was affected considerably, resulting in zero emergence (100% pupal mortality) at the highest dosages. Deformed adults emerged from pupae treated with 30% and 40% CO. The inhibitory effect of CO on the cytochrome P-450-dependent mono-oxygenase system or another oxidase system functioning at the pupal stage is proposed as a possible explanation for the failure of adult emergence.     

Biological and phylogenetic characterization of virulent Newcastle disease virus circulating in Mexico

In 2002-2003, velogenic Newcastle Disease Virus outbreaks, closely related to the Mexican isolates, were confirmed in the United States (U.S.) in southern California, Arizona, Nevada, and Texas. In this report, virulent NDVs isolated in Mexico between 1998 and 2006 were subjected to biologic characterization, using standard pathogenicity tests, and to phylogenetic analysis. Chicken embryo mean death time (MDT) test results ranged from 39.7 to 61.5 hours, and intracerebral pathogenicity index (ICPI) values were between 1.59 and 1.94, compared to a possible maximum value of 2.0. These isolates showed a dibasic amino acid motif at the fusion protein cleavage site sequence required for host systemic replication. Phylogenetic analysis indicated that the Mexican virulent NDVs belong to the class II, genotype V viruses and can be clearly divided in two groups as follows: isolates from 1998 to 2001 with close epidemiologic relationship with the latest U.S. NDV outbreaks, and phylogenetically distinct viruses, isolated from 2004 to 2006, which showed higher virulence. The assessment of the evolution of viruses from Mexico and other neighboring countries will aid in the U.S surveillance efforts for early detection of highly virulent NDV.     

Detection by RT-PCR and genetic characterization of canine distemper virus from vaccinated and non-vaccinated dogs in Argentina

RT-PCR was used to detect canine distemper virus (CDV) RNA in clotted blood from Argentine domestic dogs. The NP gene was detected in 73 out of 99 blood samples analyzed. The deduced amino acid sequence of these gene fragments showed 100% identity with the sequence of other wild-type and vaccine strains. A fragment of the hemagglutinin gene was amplified from 24 (32.9%) of the NP-RNA-positive clinical specimens. These H fragments were further analyzed by restriction fragment length polymorphism (RFLP) and sequencing. A single NdeI site was detected in all 24 wild-type strains but was absent in the vaccine strains. Phylogenetic analysis of the partial hemagglutinin amino acid sequences showed close clustering for local strains, clearly distinct from vaccine strains and other wild-type foreign CDV strains. One of the local strains, Arg 23, branched out of the root of the Argentine clade, close to the European strains, suggesting that two different pathogenic CDV genotypes are currently circulating in Argentina, one of them clearly predominant.     

Physicochemical characterization and pharmacokinetics in broiler chickens of a new recrystallized enrofloxacin hydrochloride dihydrate

Enrofloxacin, a key antimicrobial agent in commercial avian medicine, has limited bioavailability (60%). This prompted its chemical manipulation to yield a new solvate‐recrystallized enrofloxacin hydrochloride dihydrate entity (enro_C). Its chemical structure was characterized by means of mass spectroscopy, Fourier transformed infrared spectroscopy, X‐ray powder diffraction, and thermal analysis. Comparative oral pharmacokinetics (PK) of reference enrofloxacin (enro_R) and enro_C in broiler chickens after oral administration revealed noticeable improvements in key parameters and PK/PD ratios. Maximum serum concentration values were 2.61 ± 0.21 and 5.9 ± 0.42 μg/mL for enro_R and enro_C, respectively; mean residence time was increased from 5.50 ± 0.26 h to 6.20 ± 0.71 h and the relative bioavailability of enro_C was 336%. Considering C_max/MIC and AUC/MIC ratios and the MIC values for a wild‐type Escherichia coli O78/H12 (0.25 μg/mL), optimal ratios will only be achieved by enro_C (C_max/MIC = 23.6 and AUC/MIC = 197.7 for enro_C; vs. C_max/MIC = 10.4 and AUC/MIC = 78.1 for enro_R). Furthermore, enro_C may provide in most cases mutant prevention concentrations (C_max/MIC ≥ 16). Ready solubility of powder enro_C in drinking water at concentrations regularly used (0.01%) to provide an additional advantage of enro_C in the field. Further development of enro_C is warranted before it can be recommended for clinical use in veterinary medicine.     

Pedodiversity in mountainous tropical semideciduous forests of Sierra Madre del Sur,Mexico

We studied pedodiversity in mountainous tropical semideciduous forests at three key sites in Sierra Madre del Sur, Southern Mexico, at the altitudes from 600 to 1400 m a.s.l. Using detailed soil maps, we recorded the total number and the areas of soil delineations at each site, and calculated the pedodiversity indices. We found that pedodiversity increases with an increase in the absolute height within the zone of semideciduous forests, and interpreted this phenomenon as an evidence for a smaller development of erosion at higher altitudes. The areas of soil delineations reach their minimum within the intermediate part of this zone, because large delineations of strongly weathered clayey soils are developed at the higher altitudes, and large delineations of recent soils on strongly eroded surfaces are found at the lower altitudes. A conclusion about the need to take into account the particular composition of the soil cover upon the interpretation of pedodiversity data is made.