Cell proliferation and expression of connexins differ in melanotic and amelanotic canine oral melanomas

Melanoma is a malignant neoplasm occurring in several animal species, and is the most frequently found tumor in the oral cavity in dogs. Melanomas are classified into two types: melanotic and amelanotic. Prior research suggests that human amelanotic melanomas are more aggressive than their melanotic counterparts. This study evaluates the behavior of canine melanotic and amelanotic oral cavity melanomas and quantifies cell proliferation and the expression of connexins. Twenty-five melanomas (16 melanotic and 9 amelanotic) were collected from dogs during clinical procedures at the Veterinary Hospital of the School of Veterinary Medicine and Animal Science of the University of São Paulo, Brazil. After diagnosis, dogs were followed until death or euthanasia. Histopathology confirmed the gross melanotic or amelanotic characteristics and tumors were classified according to the WHO. HMB45 or Melan A immunostainings were performed to confirm the diagnosis of amelanotic melanomas. Cell proliferation was quantified both by counting mitotic figures and PCNA positive nuclei. Expressions of connexins 26 and 43 were evaluated by immunohistochemistry, qRT-PCR and Western blot. Dogs bearing amelanotic melanomas presented a shorter lifespan in comparison to those with melanotic melanomas. Cell proliferation was significantly higher in amelanotic melanomas. Expressions of Connexins 26 and 43 were significantly reduced in amelanotic melanomas. The results presented here suggest that oral cavity melanotic and amelanotic melanomas differ regarding their behavior, cell proliferation and connexin expression in dogs, indicating a higher aggressiveness of amelanotic variants.     

Effect of Cryptosporidium parvum infection on the absorptive capacity and paracellular permeability of the small intestine in neonatal calves

Cryptosporidium parvum is recognized as one of the most important pathogens causing enteritis and severe diarrhoea in calves up to 1 month of age. Although the infection may be responsible for some mortality, its impact is mainly associated with the impairment of intestinal functions and lower performance of animals. The aim of this study was to determine the effect of cryptosporidiosis on the intestinal functions in neonatal experimentally infected Holstein calves. Absorption tests with d-xylose and retinyl-palmitate, and the lactulose/mannitol test of intestinal permeability were simultaneously performed in 1-week intervals from challenge to full recovery. In infected animals, reduced intestinal absorptive capacity for both D-xylose and retinyl-palmitate was observed on day 7 post-infection (p.i.). At the same time, a more than 100% elevation of intestinal permeability was observed in the infected calves. All intestinal functions, except absorption of retinyl-palmitate, were significantly affected and changes were detected up to day 14 p.i. In contrast, results of all tests obtained on day 21 p.i. suggest full recovery of the infected intestine. Significantly, growth of the calves which had recovered from cryptosporidiosis was still affected between days 14 and 21 p.i.     

Detection of organophosphate and pyrethroid resistance alleles in Czech Leptinotarsa decemlineata (Coleoptera: Chrysomelidae) populations by molecular methods

BACKGROUND: Pyrethroids and organophosphates are the most frequently used insecticides for Colorado potato beetle, Leptinotarsa decemlineata (Say), control in the Czech Republic. Based on molecular methods, organophosphate and pyrethroid resistance alleles have been detected in samples from three sites. The accuracy of restriction fragment length polymorphism (RFLP) and bidirectional polymerase chain reaction amplification of specific alleles (Bi‐PASA) for detection of resistance alleles is compared. RESULTS: Leptinotarsa decemlineata from three sites showed higher frequencies of resistance alleles to organophosphates than to pyrethroids. The rates of occurrence of individuals homozygous resistant (RR) to pyrethroids ranged from 20.0 to 22.9%, while the rates of occurrence of individuals RR to organophosphates ranged from 52.9 to 66.7%. The incidences of individuals with resistance alleles to both organophosphates and pyrethroids ranged from 8.6 to 13.6%. No relationship was found between incidence of the pyrethroid resistance allele and site, while incidence of the organophosphate resistance allele differed significantly according to site. CONCLUSION: Both RFLP and Bi‐PASA were suitable for detecting resistance alleles to pyrethroids, and in most cases also for detecting resistance alleles to organophosphates. In contrast to Bi‐PASA, RFLP was also suitable for samples with lower DNA quality when testing for the resistance allele to pyrethroids. On the other hand, RFLP was not as accurate as Bi‐PASA in detection of the organophosphate resistance allele. Copyright © 2010 Society of Chemical Industry     

Effects of marine climate variability on the relative abundance of Lutjanus purpureus (POEY, 1866) on the Amazon Continental Shelf

The aim of this study was to analyze the effects of climate and marine variability on the catches of Lutjanus purpureus in three sectors of the Amazon Continental Shelf. Remote sensing data were compared with landings (CPUE) between 1997 and 2007 and analyzed for partial influence obtained through the Generalized Additive Model. Additionally, significant variables were analyzed through Wavelet Cross-Spectrum, and periods of high correlation between variables in space and time were identified. The results indicated a high coherence between catch per unit effort (CPUE) and environmental variables on an interannual scale, suggesting that Red Snapper fishing is mainly influenced by the seasonal effects of oceanographic variables. These results suggest that fluctuations in Red Snapper catches between 1997 and 2007 may be the result of natural processes that occur in the Amazon Continental Shelf every year in synergy with environmental variables, which are also indirectly affected by the El Niño–Southern Oscillation (ENSO) pattern.     

SIRT1-dependent modulation of methylation and acetylation of histone H3 on lysine 9 (H3K9) in the zygotic pronuclei improves porcine embryo development

Background: The histone code is an established epigenetic regulator of early embryonic development in mammals.The lysine residue K9 of histone H3(H3 K9) is a prime target of SIRT1, a member of NAD+-dependent histone deacetylase family of enzymes targeting both histone and non-histone substrates. At present, little is known about SIRT1-modulation of H3 K9 in zygotic pronuclei and its association with the success of preimplantation embryo development. Therefore, we evaluated the effect of SIRT1 activity on H3 K9 methylation and acetylation in porcine zygotes and the significance of H3 K9 modifications for early embryonic development.Results: Our results show that SIRT1 activators resveratrol and BML-278 increased H3 K9 methylation and suppressed H3 K9 acetylation in both the paternal and maternal pronucleus. Inversely, SIRT1 inhibitors nicotinamide and sirtinol suppressed methylation and increased acetylation of pronuclear H3 K9. Evaluation of early embryonic development confirmed positive effect of selective SIRT1 activation on blastocyst formation rate(5.2 ± 2.9% versus 32.9 ± 8.1% in vehicle control and BML-278 group, respectively; P ≤ 0.05). Stimulation of SIRT1 activity coincided with fluorometric signal intensity of ooplasmic ubiquitin ligase MDM2, a known substrate of SIRT1 and known limiting factor of epigenome remodeling.Conclusions: We conclude that SIRT1 modulates zygotic histone code, obviously through direct deacetylation and via non-histone targets resulting in increased H3 K9 me3. These changes in zygotes lead to more successful pre-implantation embryonic development and, indeed, the specific SIRT1 activation due to BML-278 is beneficial for in vitro embryo production and blastocyst achievement.     

Distinguishing regional and local sources of pollution by trace metals and magnetic particles in fluvial sediments of the Morava River, Czech Republic

Purpose

Regional contamination of southern Moravia (SE part of the Czech Republic) by trace metals and magnetic particles during the twentieth century was quantified in fluvial sediments of the Morava River. The influence of local pollution sources on regional contamination of the river sediments and the effect of sampling site heterogeneity were studied in sediment profiles with different lithologies.

Materials and methods

Hundreds of sediment samples were obtained from regulated channel banks and naturally inundated floodplains and proxy elemental analyses were carried out by energy dispersive X-ray fluorescence spectroscopy (ED XRF) and further calibrated by inductively coupled plasma mass spectrometry (ICP MS). Magnetic susceptibility was determined as a proxy for industrial contamination. The age model for the floodplain sediments was established from ¹³⁷Cs and ²¹⁰Pb dating. Trace metal contamination was assessed by establishing the lithological background values from floodplain profiles and calculating enrichment factors (EF) of trace metals (i.e. Pb, Zn, Cu) and magnetic susceptibility for the entire study area.

Results and discussion

Channel sediments are unsuitable for the reconstruction of historical regional contamination due to their lithological heterogeneity and the “chaotic” influence of local sources of contamination, as well as the possibility of geochemical mobility of pollutants. On the other hand, sediments from regulated river banks qualitatively reflected the actual, local contamination of the river system.

Conclusions

This approach allowed us to distinguish the influence of local sources of contamination by comparison with more spatially averaged contamination signals from distal floodplain profiles. The studied area is weakly contaminated (EF ～1–2), while individual sediment strata from regulated channel banks reflect local sources of contamination and contain up to several times higher concentrations of trace metals.     

Division of labor and resource management in eastern Pará, Brazil

Peasants of the northeastern Pará cultivate cassava (manioc esculenta) using shifting cultivation. This paper discusses some factors in support of cassava production, even though it has some negative environmental impacts. These factors are the importance of cassava in the region's history, dietary traditions, and the cooperative labor systems employed in its cultivation and processing. This revised version was published online in June 2006 with corrections to the Cover Date.     

Osteogenic potential of mesenchymal cells derived from canine umbilical cord matrix co-cultured with platelet-rich plasma and demineralized bone matrix

Canine mesenchymal cells (MSCs) derived from Wharton''s jelly were co-cultured, then supplemented or not supplemented with platelet rich plasma (PRP) and demineralized bone matrix (DBM) to verify osteogenic differentiation. Osteoblastic differentiation followed by mineralized bone matrix production was found to be significantly higher (p < 0.05) when MSCs were associated with PRP/DBM in culture after 14-21-days of induction. Osteopontin and osteocalcin gene expression were significantly superior (p < 0.05) under the same culture conditions after 21 days of observation. In conclusion, addition of PRP to DBM co-cultured with MSCs successfully induced osteogenesis in vitro.