Flow Cytometry Identification of X- and Y-Chromosome-Bearing Goat Spermatozoa

Flow cytometric sorting technology was used to measure the difference in DNA content between X- and Y-chromosome-bearing spermatozoa in bucks. Spermatozoa were analysed by flow cytometry to characterize X- and Y-chromosome-bearing sperm populations and to quantify the DNA difference between them. Two symmetrical, overlapping and clearly separated peaks, corresponding to X- and Y-bearing spermatozoa, were detected. The difference in fluorescence intensity between the peaks was 4.4 +/- 0.03% without any significant inter- or intra-animal variations. Therefore, the identification and selection of high-purity samples of sperm populations for sex sorting is easier in bucks compared with other domestic species.     

Spatial Modeling of Tropical Deforestation Using Socioeconomic and Biophysical Data

Tropical forests are disappearing at an alarming rate. In Central America, a hectare of forest is cleared for agriculture every 5 min. This study was conducted in a forested 4,000 ha watershed of central Honduras to find deforestation causes based on socio-economic characteristics of population. First, a multitemporal analysis of Landsat TM imagery was conducted to determine deforestation rates and agricultural–forest boundaries. A GIS buffer procedure allowed determining which households were at the deforestation front and which households were located at the rest of the area (control). GIS techniques were used to extract biophysical information such as slope, elevation, land cover, temperature, precipitation, etc. Then, we set up a data base with more than 50 socioeconomic variables (level of education, income, children per family, major economic activity, use of conservation practices, etc.). Around 500 households, distributed all over the watershed, were visited, interviewed and GPS-located. A multivariate statistical analysis allowed an exploratory analysis to eliminate non useful and redundant variables and then to determine what variables appear to be important predictors of deforestation behavior among rural families. A resulting logistic regression model showed that household with lower annual income heads and with less use of conservation practices were more statistically prone to clear the forest (α = 0.001). The study uncovered the complexity of this problem and confirmed the need of using GIS–remote sensing techniques to combine socioeconomic and environmental data in several time–space dimensions to find the causes and trends of tropical deforestation.     

Immunohistochemical differentiation of reactive from malignant mesothelium as a diagnostic aid in canine pericardial disease

Objectives

To develop a provisional immunohistochemistry panel for distinguishing reactive pericardium, atypical mesothelial proliferation and mesothelioma in dogs.

Materials and Methods

Archived pericardial biopsies were subject to haematoxylin and eosin staining, immunohistochemistry for cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Samples were scored for intensity and number of cells stained.

Results

Ten biopsies of reactive mesothelium, 17 of atypical mesothelial proliferation, 26 of mesothelioma and five of normal pericardium were identified on the basis of haematoxylin and eosin staining. Cytokeratin and vimentin were expressed in all biopsies, confirming mesothelial origin. Normal pericardial samples had the lowest scores for insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Mesothelioma and atypical proliferative samples were similar to each other, with higher scores for insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 than the reactive samples. Desmin staining was variable. Insulin‐like growth factor II mRNA‐binding protein 3 was the best to distinguish between disease groups.

Clinical Significance

An immunohistochemistry panel of cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 could provide superior information compared with haematoxylin and eosin staining alone in the diagnosis of cases of mesothelial proliferation in canine pericardium, but further validation is warranted.     

A high-resolution radiation hybrid map of the human genome draft sequence

We have constructed a physical map of the human genome by using a panel of 90 whole-genome radiation hybrids (the TNG panel) in conjunction with 40,322 sequence-tagged sites (STSs) derived from random genomic sequences as well as expressed sequences. Of 36,678 STSs on the TNG radiation hybrid map, only 3604 (9.8%) were absent from the unassembled draft sequence of the human genome. Of 20,030 STSs ordered on the TNG map as well as the assembled human genome draft sequence and the Celera assembled human genome sequence, 36% of the STSs had a discrepant order between the working draft sequence and the Celera sequence. The TNG map order was identical to one of the two sequence orders in 60% of these discrepant cases.     

Effect of Fucoidan on the Mitochondrial Membrane Potential (ΔΨm) of Leukocytes from Patients with Active COVID-19 and Subjects That Recovered from SARS-CoV-2 Infection

Fucoidan is a polysaccharide obtained from marine brown algae, with anti-inflammatory, anti-viral, and immune-enhancing properties, thus, fucoidan may be used as an alternative treatment (complementary to prescribed medical therapy) for COVID-19 recovery. This work aimed to determine the ex-vivo effects of treatment with fucoidan (20 µg/mL) on mitochondrial membrane potential (ΔΨm, using a cationic cyanine dye, 3,3′-dihexyloxacarbocyanine iodide (DiOC₆(3)) on human peripheral blood mononuclear cells (HPBMC) isolated from healthy control (HC) subjects, COVID-19 patients (C-19), and subjects that recently recovered from COVID-19 (R1, 40 ± 13 days after infection). In addition, ex-vivo treatment with fucoidan (20 and 50 µg/mL) was evaluated on ΔΨm loss induced by carbonyl cyanide 3-chlorophenylhydrazone (CCCP, 150 µM) in HPBMC isolated from healthy subjects (H) and recovered subjects at 11 months post-COVID-19 (R2, 335 ± 20 days after infection). Data indicate that SARS-CoV-2 infection induces HPBMC loss of ΔΨm, even 11 months after infection, however, fucoidan promotes recovery of ΔΨm in PBMCs from COVID-19 recovered subjects. Therefore, fucoidan may be a potential treatment to diminish long-term sequelae from COVID-19, using mitochondria as a therapeutic target for the recovery of cellular homeostasis.     

Impact of water content on the solubilisation of arabinoxylan during xylanase treatment of wheat bran

Arabinoxylan (AX) has a major impact on the functional properties of wheat bran, and it has been shown that technological properties of bran can be improved by using endoxylanases. Enzymatic treatments are typically conducted at high water content. However, in industrial applications, low water content may be advantageous, especially when targeting dry end products. The aim of the study was to examine the impact of water content, ranging from 20 to 90%, on the efficiency of endoxylanase treatment of wheat bran. Interestingly, AX solubilisation was highest at the water contents of 40 and 90%. At water contents 50–80%, AX solubilisation was lower than at 40 and 90%. Furthermore, at low water content, less depolymerisation was detected. At water content of 40%, the bran-water mixture was transformed from powder-like into compact mass. Probably the compact consistency of the material enhanced AX solubilisation by increased breakdown of bran cell walls due to shear forces or via enhanced enzyme binding to the substrate. The results show that solubilisation of bran AX can also be efficiently performed at low water content.     

A new acylated quercetin glycoside and other secondary metabolites from Helleborus foetidus

A new acylated flavonol glycoside, quercetin 3-O-(2-trans-caffeoyl)-alpha-L-arabinopyranosyl-(1-->2)-beta-D-glucopyranoside (1), together with the known 25R,26-[(beta-D-glucopyranosyl)oxy]-22alpha-hydroxy-5beta-furostan-3-beta-yl O-alpha-l-rhamnopyranosyl-(1-->4)-beta-D-glucopyranoside (2), anemonin (3), beta-D-glucosyl-p-hydroxyphenylethyl alcohol (4), and 1-beta-O-caffeoyl-D-glucose (5) were isolated from Helleborus foetidus leaves and identified on the basis of detailed spectral analysis.