A promising new ELISA diagnostic test for cattle babesiosis based on Babesia bigemina Apical Membrane Antigen-1.

Babesiosis due to Babesia bigemina is a relevant tick‑borne disease, affecting cattle worldwide. Many surface proteins of the pathogen including the Apical Membrane Antigen 1 (AMA‑1) ‑ have been analysed for vaccine and diagnostic purposes. This study focused on B. bigemina AMA‑1 and on its use for the assessment of diagnostic tests. After bioinformatic analyses, AMA‑1 codifying region was amplified and cloned into an expression vector used to induce protein synthesis in Escherichia coli cells. AMA‑1 was purified by affinity chromatography and used to set up the best condition for an ELISA protocol. Bovine field sera positive to B. bigemina were used to evaluate the presence of anti‑AMA‑1 antibodies. In order to verify the assay specificity, sera positive to Babesia bovis or to the piroplasm Theileria annulata were also included. Significant differences were obtained between sera negative to both B. bigemina and B. bovis and samples positive to B. bigemina, to B. bovis or to both pathogens. No significant reaction was observed with T. annulata positive sera. The results showed that AMA‑1 protein is suitable to be used as antigen in diagnostic assays for babesiosis diagnosis in cattle, as it does not show any cross reaction with anti-T. annulata antibodies

Perturbations of the local gravity field due to mass distribution on precise measuring instruments: a numerical method applied to a cold atom gravimeter

We present a numerical method, based on a FEM simulation, for the determination of the gravitational field generated by massive objects, whatever geometry and space mass density they have. The method was applied for the determination of the self gravity effect of an absolute cold atom gravimeter which aims at a relative uncertainty of 10-9. The deduced bias, calculated with a perturbative treatment, is finally presented. The perturbation reaches (1.3 \pm 0.1) \times 10-9 of the Earth's gravitational field.Comment: 12 pages, 7 figure

Self-attraction effect and correction on three absolute gravimeters

The perturbations of the gravitational field due to the mass distribution of an absolute gravimeter have been studied. The so called Self Attraction Effect (SAE) is crucial for the measurement accuracy, especially for the International Comparisons, and for the uncertainty budget evaluation. Three instruments have been analysed: MPG-2, FG5-238 and IMPG-02. The SAE has been calculated using a numerical method based on FEM simulation. The observed effect has been treated as an additional vertical gravity gradient. The correction (SAC) to be applied to the computed g value has been associated with the specific height level, where the measurement result is typically reported. The magnitude of the obtained corrections is of order 1E-8 m/s2.Comment: 14 pages, 8 figures, submitted to Metrologi

Fluorescence Spectroscopy for the Diagnosis of Endometritis in the Mare

By exploiting the PMN property to produce high quantities of oxygen peroxide to neutralize pathogens, the oxygen peroxide content of uterine cells was measured to diagnose endometritis. After preliminary in vitro studies in which endometrial cells from slaughtered mares were mixed with leukocytes from peripheral blood, endometrial samples were collected by uterine flushing from mares before insemination. Staining endometrial cells with H2DCF-DA was combined with hydroethidine to normalize the fluorescence intensity with the cellular content of the sample. Stained cell smears were assumed as the gold standard of endometritis, and based on this assay, the samples were considered positive (C+) and negative (C-) for endometritis. The amount and the turbidity of fluid recovered by uterine flushing were significantly (p < 0.01) higher in C+ than in C-. Moreover, the oxygen peroxide content of the endometrial cells was significantly higher in the C+ than in the C- group (6.31 ± 1.92 vs. 3.12 ± 1.26, p = 0.001). Using the value of 4.4 as the cutoff level of this fluorescence cytology assay, it was found that only one C- sample exceeded the cutoff level (false positives = 7.7%) while three C+ samples showed values below the cutoff level (false negative = 11.5%)

Environmentally Friendly Sunscreens: Mechanochemical Synthesis and Characterization of β-CD Inclusion Complexes of Avobenzone and Octinoxate with Improved Photostability

We report on the mechanochemical synthesis of inclusion complexes obtained by reacting β-cyclodextrin (β-CD) with two widely used sunscreens, namely, avobenzone (AVO) and octinoxate (OCT). Formation of crystalline inclusion complexes was confirmed via a combination of solid-state techniques, including X-ray diffraction (XRD), Raman, and ATR-FTIR spectroscopies. A new, metastable polymorph of avobenzone was also isolated and characterized. NMR spectroscopy and thermal analyses (TGA and DSC) allowed us to evaluate the host/guest ratio and the water content (ca. 8H2O) in crystalline (β-CD)2·AVO and (β-CD)3·OCT2. Photodegradation of the two sunscreens upon inclusion in the hydrophobic cavity of β-CD was evaluated in solution via mass spectrometry (ESI-MS) and UV-vis spectroscopy and found to be sharply reduced. All findings indicate that the inclusion of AVO and OCT in β-CD might represent a viable route for the preparation of environmentally friendly sunscreens with improved photostability to be used in formulations of sun creams

Isotopic and Microcanonical Temperatures in Nuclear Multifragmentation

A systematic comparison of different isotopic temperatures with the thermodynamical temperature of a multifragment system is made on the basis of the Statistical Multifragmentation Model. It is demonstrated that isotopic temperatures are strongly affected by the secondary decays of hot primary fragments and the population of particle-stable excited states in final fragments. The He-Li temperatures, measured recently by the ALADIN group, are reproduced fairly well both as a function of excitation energy and bound charge. Our analysis confirms the anomaly in the nuclear caloric curve.Comment: 10 pages in LaTeX, 3 ps figures, accepted for publication in Phys. Rev.

Cytotoxic activity of organotin(IV) derivatives with triazolopyrimidine containing exocyclic oxygen atoms

In this study cytotoxicity of organotin(IV) compounds with 1,2,4-triazolo[1,5-a]pyrimidines, Me3Sn(5tpO) (1), n-Bu3Sn(5tpO) (2), Me3Sn(mtpO) (3), n-Bu3Sn(mtpO) (4), n-Bu3Sn(HtpO2) (5), Ph3Sn(HtpO2) (6) where 5HtpO = 4,5-dihydro-5-oxo-[1,2,4]triazolo-[1,5-a]pyrimidine, HmtpO = 4,7-dihydro-5-methyl-7-oxo-[1,2,4]triazolo-[1,5-a]pyrimidine, and H2tpO2 = 4,5,6,7-tetrahydro-5,7-dioxo-[1,2,4]triazolo-[1,5-a]-pyrimidine, was assessed on three different human tumor cell lines: HCT-116 (colorectal carcinoma), HepG2 (hepatocarcinoma) and MCF-7 (breast cancer). While 1 and 3 were inactive, compounds 2, 4, 5 and 6 inhibited the growth of the three tumor cell lines with IC50 values in the submicromolar range and showed high selectivity indexes towards the tumor cells (SI > 90). The mechanism of cell death triggered by the organotin(IV) derivatives, investigated on HCT-116 cells, was apoptotic, as evident from the externalization of phosphatidylserine to the cell surface, and occurred via the intrinsic pathway with fall of mitochondrial inner membrane potential and production of reactive oxygen species. While compound 6 arrested the cell progression in the G2/M cell cycle phase and increased p53 and p21 levels, compounds 2, 4 and 5 blocked cell duplication in the G1 phase without affecting the expression of either of the two tumor suppressor proteins. Compounds 1 and 2 were also investigated using single crystal X-ray diffraction and found to be, in both cases, coordination polymers forming 1 D chains based on metal-ligand interactions. Interestingly, for n-Bu3Sn(5tpO)(2) H-bonding interactions between 5tpO− ligands belonging to adjacent chains were also detected that resemble the “base-pairing” assembly and could be responsible for the higher biological activity compared to compound 1. In addition, they are the first example of bidentate N(3), O coordination for the 5HtpO ligand on two adjacent metal atoms

Genetic Correlates of Brain Aging on MRI and Cognitive Test Measures: A Genome-Wide Association and Linkage Analysis in the Framingham Study

BACKGROUND: Brain magnetic resonance imaging (MRI) and cognitive tests can identify heritable endophenotypes associated with an increased risk of developing stroke, dementia and Alzheimer's disease (AD). We conducted a genome-wide association (GWA) and linkage analysis exploring the genetic basis of these endophenotypes in a community-based sample. METHODS: A total of 705 stroke- and dementia-free Framingham participants (age 62 +9 yrs, 50% male) who underwent volumetric brain MRI and cognitive testing (1999–2002) were genotyped. We used linear models adjusting for first degree relationships via generalized estimating equations (GEE) and family based association tests (FBAT) in additive models to relate qualifying single nucleotide polymorphisms (SNPs, 70,987 autosomal on Affymetrix 100K Human Gene Chip with minor allele frequency ≥ 0.10, genotypic call rate ≥ 0.80, and Hardy-Weinberg equilibrium p-value ≥ 0.001) to multivariable-adjusted residuals of 9 MRI measures including total cerebral brain (TCBV), lobar, ventricular and white matter hyperintensity (WMH) volumes, and 6 cognitive factors/tests assessing verbal and visuospatial memory, visual scanning and motor speed, reading, abstract reasoning and naming. We determined multipoint identity-by-descent utilizing 10,592 informative SNPs and 613 short tandem repeats and used variance component analyses to compute LOD scores. RESULTS: The strongest gene-phenotype association in FBAT analyses was between SORL1 (rs1131497; p = 3.2 × 10-6) and abstract reasoning, and in GEE analyses between CDH4 (rs1970546; p = 3.7 × 10-8) and TCBV. SORL1 plays a role in amyloid precursor protein processing and has been associated with the risk of AD. Among the 50 strongest associations (25 each by GEE and FBAT) were other biologically interesting genes. Polymorphisms within 28 of 163 candidate genes for stroke, AD and memory impairment were associated with the endophenotypes studied at p < 0.001. We confirmed our previously reported linkage of WMH on chromosome 4 and describe linkage of reading performance to a marker on chromosome 18 (GATA11A06), previously linked to dyslexia (LOD scores = 2.2 and 5.1). CONCLUSION: Our results suggest that genes associated with clinical neurological disease also have detectable effects on subclinical phenotypes. These hypothesis generating data illustrate the use of an unbiased approach to discover novel pathways that may be involved in brain aging, and could be used to replicate observations made in other studies.National Institutes of Health National Center for Research Resources Shared Instrumentation grant (ISI0RR163736-01A1); National Heart, Lung, and Blood Institute's Framingham Heart Study (N01-HC-25195); National Institute of Aging (5R01-AG08122, 5R01-AG16495); National Institute of Neurological Disorders and Stroke (5R01-NS17950