Genetic divergence in Northamerican freshwater planarians of the Dugesia dorotocephala group (Turbellaria, Tricladida, Paludicola)

The genetic differentiation between the members of the Dugesia (Girardia) dorotocephala group was analyzed by means of multilocus electrophoresis, and comared to that of another planarian secies, D. tahitiensis, also belonging to the subgenus Girardia. The species examined were: D. dorotocephala s.s (2n = 16), D. arizonensis (2n = 8), D. jenkinsae (2n = 8), and the above mentioned D. tahitiensis (2n = 16). The former three species inhabit North America, and show different proportion of fissiparous and sexual individuals; the latter species inhabits Polynesia and is fully asexual. A total of 11 enzyme loci were genetically analyzed: Mdh-1, Mdh-2, Zdh-1, Idh-2, G3pdh, Got-1, Ck, Pgm-2, Ada, Mpi, and Gpi. Low values of observed mean heterozygosity per locus (Ho) were found in the populations studied, ranging from 0 to 0.18 (average 0.08. In asexual populations (except that of D. tahitiensis) fixed heterozygosity was observered in all the individuals for 1 or 2 loci. The genetic divergence between the species examined is very high, with many loci showing discriminating alleles in different taxa (Nei's genetic distance varies from 0.871 to 1.759). The populations of D. dorotocehala s.s., on the contrary, appear to be genetically quite homogenous average D= 0.019), and the genetic distance values are apparently unrelated to their geographic location and to their way of reproduction. The genetic distance between D. tahitiensis, a species not included in the D. dorotocephala group and D. dorotocephala s.s. is 1.314 and hence similar to the D value between two members of;he dorotocephala group: D. dorotocephala and D. jenkinsae (D = 1.303). The genetic relationships among the populations studied were established by UPGMA cluster analysis and multidimensional scaling. The descendence of the North American species with 2n = 8 from a dorotocephala-like ancestor with 2n = 16 is considered. It is suggested that the latter, as well as a tahitiensis-like line, also having 2n = 16, have originated from a common ancestor by geographic isolation

Isolation of intermediate valence hybrids between ferrous and methemoglobin at subzero temperatures.

Quenching a hemoglobin solution partially saturated with carbon monoxide into a hydro-organic solvent containing ferricyanide will produce under suitable conditions a population of partially oxidized and CO-bound hemoglobin molecules. Since each Fe3+ heme carries one extra charge, it should be possible, in theory, to resolve the spectrum of intermediate compounds between hemoglobin and carbon monoxide, which was originally present in solution. In this study we report: 1) the development of a simple and rapid method to quench aqueous hemoglobin solutions into a hydro-organic solvent at subzero temperatures; 2) the determination of suitable experimental conditions to isolate valence hybrids between carbonmonoxy- and methemoglobin by isoelectric focusing at temperatures as low as -25 degrees C; and 3) the identification and isolation of all valence hybrids of different charge between carbonmonoxy- and methemoglobin

Heat shock protein 90 is associated with hyperplasia and neoplastic transformation of canine prostatic epithelial cells

Heat shock protein 90 (HSP90) is a molecular chaperone that regulates critical signalling proteins of cancer development and progression. Abnormal levels of HSP90 have been observed in human prostatic carcinoma (PC), with prognostic and therapeutic implications. Since spontaneously arising canine PC is a valuable model for the human disease, the aim of this study was to evaluate the immunohistochemical expression of HSP90 in two normal canine prostates, 17 canine prostates with benign prostatic hyperplasia (BPH) and five canine prostates with PC. HSP90 was expressed in the cytoplasm of epithelial cells in all samples, with a significant increase in labelled cells in PCs. Nuclear labelling was observed occasionally in normal tissue, but was increased in BPH and PC. HSP90 immunoreactivity in preneoplastic lesions (proliferative inflammatory atrophy and prostatic intraepithelial neoplasia) was similar to that in PCs. Increased HSP90 expression in canine PCs suggests the involvement of this molecule in carcinogenesis and tumour progression, supporting HSP90 as a potential target for therapeutic intervention

Isolation of intermediate compounds between hemoglobin and carbon monoxide.

A human hemoglobin solution partially saturated with carbon monoxide was rapidly quenched at -25 degrees C into a hydro-organic buffer containing ferricyanide. Under the experimental conditions of pH, ionic strength, and buffer composition used in this work, it was found that the deoxy hemes were rapidly transformed into their met form, whereas practically no carbon monoxide-bound hemes were oxidized before the separation of the mixture from the oxidizing agent. As a preliminary step to the analysis of the resulting solution, carbonylhemoglobin solutions partially oxidized with ferricyanide were studied by isoelectric focusing at -25 degrees C under identical conditions. The relative position in the gel of all nine possible valence hybrids was established as follows (going from the anodic to the cathodic side of the gel) alpha CO2 beta CO2, (alpha CO beta +)(alpha CO beta CO) (alpha CO beta CO), (alpha CO2 beta +2), (alpha + beta CO), (alpha + beta +)-(alpha CO beta CO), (alpha + beta +)(alpha CO beta +), (alpha +2 beta CO2), (alpha + beta +)(alpha + beta CO), alpha +2 beta +2. When carbonylhemoglobin and methemoglobin were mixed in equal proportion at -25 degrees C and then analyzed by isoelectric focusing at the same temperature, it was found that the contribution of valence hybrids other than alpha CO2 beta CO2 and alpha +2 beta +2 to the total amount of hemoglobin in the gel was no more than 6%. When carbonylhemoglobin and deoxyhemoglobin were mixed in the same proportion and incubated at 20 degrees C so to allow the redistribution of the carbon monoxide molecules between all possible binding sites to occur, a substantially higher amount of valence hybrids, derived from the oxidation of intermediate compounds of hemoglobin with carbon monoxide, was found. The isoelectric focusing separation indicated the presence in the original solution of intermediate species other than carbonylhemoglobin and deoxyhemoglobin at a concentration of about 10% of the total

A strontium isoscape of Italy for provenance studies

We present a novel database of biological and geological 87Sr/86Sr values (n = 1920) from Italy, using literature data and newly analysed samples, for provenance purposes. We collected both bioavailable and non-bioavailable (i.e. rocks and bulk soils) data to attain a broader view of the Sr isotope variability of the Italian territory. These data were used to build isotope variability maps, namely isoscapes, through Kriging interpolations. We employed two different Kriging models, namely Ordinary Kriging and Universal Kriging, with a geolithological map of Italy categorized in isotope classes as external predictor. Model performances were evaluated through a 10-fold cross validation, yielding accurate 87Sr/86Sr predictions with root mean squared errors (RMSE) ranging between 0.0020 and 0.0024, dependent on the Kriging model and the sample class. Overall, the produced maps highlight a heterogeneous distribution of the 87Sr/86Sr across Italy, with the highest radiogenic values (>0.71) mainly localized in three areas, namely the Alps (Northern Italy), the Tuscany/Latium (Central Italy) and Calabria/Sicily (Southern Italy) magmatic/metamorphic terrains. The rest of the peninsula is characterized by values ranging between 0.707 and 0.710, mostly linked to sedimentary geological units of mixed nature. Finally, we took advantage of the case study of Fratta Polesine, to underscore the importance of choosing appropriate samples when building the local isoscape and of exploring different end-members when interpreting the local Sr isotope variability in mobility and provenance studies. Our user-friendly maps and database are freely accessible through the Geonode platform and will be updated over time to offer a state-of-the-art reference in mobility and provenance studies across the Italian landscape

Secondary dentin formation mechanism: The effect of attrition

Human dentin consists of a primary layer produced during tooth formation in early child-hood and a second layer which first forms upon tooth eruption and continues throughout life, termed secondary dentin (SD). The effect of attrition on SD formation was considered to be confined to the area subjacent to attrition facets. However, due to a lack of three‐dimensional methodologies to demonstrate the structure of the SD, this association could not be determined. Therefore, in the current study, we aimed to explore the thickening pattern of the SD in relation to the amount of occlusal and interproximal attrition. A total of 30 premolars (50–60 years of age) with varying attrition rates were evaluated using micro‐computerized tomography. The results revealed thickening of the SD below the cementoenamel junction (CEJ), mostly in the mesial and distal aspects of the root (p < 0.05). The pattern of thickening under the tooth cervix, rather than in proximity to attrition facets, was consistent regardless of the attrition level. The amount of SD thickening mildly corre-lated with occlusal attrition (r = 0.577, p < 0.05) and not with interproximal attrition. The thickening of the SD below the CEJ coincided with previous finite element models, suggesting that this area is mostly subjected to stress due to occlusal loadings. Therefore, we suggest that the SD formation might serve as a compensatory mechanism aimed to strengthen tooth structure against deflection caused by mechanical loading. Our study suggests that occlusal forces may play a significant role in SD formation

3D enamel thickness in Neandertal and modern human permanent canines

Enamel thickness figures prominently in studies of human evolution, particularly for taxonomy, phylogeny, and paleodietary reconstruction. Attention has focused on molar teeth, through the use of advanced imaging technologies and novel protocols. Despite the important results achieved thus far, further work is needed to investigate all tooth classes. We apply a recent approach developed for anterior teeth to investigate the 3D enamel thickness of Neandertal and modern human (MH) canines. In terms of crown size, the values obtained for both upper and lower unworn/slightly worn canines are significantly greater in Neandertals than in Upper Paleolithic and recent MH. The 3D relative enamel thickness (RET) is significantly lower in Neandertals than in MH. Moreover, differences in 3D RET values between the two groups appear to decrease in worn canines beginning from wear stage 3, suggesting that both the pattern and the stage of wear may have important effects on the 3D RET value. Nevertheless, the 3D average enamel thickness (AET) does not differ between the two groups. In both groups, 3D AET and 3D RET indices are greater in upper canines than in lower canines, and overall the enamel is thicker on the occlusal half of the labial aspect of the crown, particularly in MH. By contrast, the few early modern humans investigated show the highest volumes of enamel while for all other components of 3D enamel, thickness this group holds an intermediate position between Neandertals and recent MH. Overall, our study supports the general findings that Neandertals have relatively thinner enamel than MH (as also observed in molars), indicating that unworn/slightly worn canines can be successfully used to discriminate between the two groups. Further studies, however, are needed to understand whether these differences are functionally related or are the result of pleiotropic or genetic drift effects

P-Glycoprotein and Breast Cancer Resistance Protein in Canine Inflammatory and Noninflammatory Grade III Mammary Carcinomas

P-glycoprotein (P-gp/ABCB1) and breast cancer resistance protein (BCRP/ABCG2) expression are frequently related to multidrug resistance (MDR) in neoplastic cells. Canine inflammatory and grade III noninflammatory mammary carcinomas (IMC and non-IMC) are aggressive tumors that could benefit from chemotherapy. This study describes the immunohistochemical detection of P-gp and BCRP in 20 IMCs and 18 non-IMCs from dogs that had not received chemotherapy. Our aim was to determine if P-gp and BCRP expression was related to the \u201cinflammatory\u201d phenotype, to establish a basis for future studies analyzing the response to chemotherapy in dogs with highly malignant mammary cancer. Immunolabeling was primarily membranous for P-gp with a more intense labeling in emboli, and immunolabeling was membranous and cytoplasmic for BCRP. P-gp was expressed in 17 of 20 (85%) IMCs compared to 7 of 18 (39%) non-IMCs (P = 0.006). BCRP was expressed within emboli in 15 of 19 (79%) emboli in IMC, 12 of 15 (80%) primary IMCs, and 12 of 18 (67%) non-IMCs, without statistically significant differences (P >.05). All IMCs and 67% of non-IMCs expressed at least 1 of the 2 transporters, and 63% (12/19) of IMCs and 39% (7/18) of non-IMCs expressed both P-gp and BCRP. P-gp and BCRP evaluation might help select patients for chemotherapy. P-gp, expressed in a significantly higher percentage of IMCs vs non-IMCs, might play a specific role in the chemoresistance of IMC

Deletion in the EVC2 gene causes chondrodysplastic dwarfism in Tyrolean grey cattle

During the summer of 2013 seven Italian Tyrolean Grey calves were born with abnormally short limbs. Detailed clinical and pathological examination revealed similarities to chondrodysplastic dwarfism. Pedigree analysis showed a common founder, assuming autosomal monogenic recessive transmission of the defective allele. A positional cloning approach combining genome wide association and homozygosity mapping identified a single 1.6 Mb genomic region on BTA 6 that was associated with the disease. Whole genome re-sequencing of an affected calf revealed a single candidate causal mutation in the Ellis van Creveld syndrome 2 (EVC2) gene. This gene is known to be associated with chondrodysplastic dwarfism in Japanese Brown cattle, and dwarfism, abnormal nails and teeth, and dysostosis in humans with Ellis-van Creveld syndrome. Sanger sequencing confirmed the presence of a 2 bp deletion in exon 19 (c.2993_2994ACdel) that led to a premature stop codon in the coding sequence of bovine EVC2, and was concordant with the recessive pattern of inheritance in affected and carrier animals. This loss of function mutation confirms the important role of EVC2 in bone development. Genetic testing can now be used to eliminate this form of chondrodysplastic dwarfism from Tyrolean Grey cattle