Effect of somatotropin on changes in milk-production and composition during coliform mastitis in periparturient cows.

The potential protective and therapeutic effects of bST during coliform mastitis in periparturient cows were evaluated. In a first study, 19 cows, classified as moderate or severe responders based on the respiratory burst activity of blood neutrophils, were treated with recombinant bST or vehicle 48 h after intramammary inoculation of Escherichia coli. Clinical status and changes in milk production and composition were compared in the four groups. In a second study, 8 cows received bST or vehicle 7 d before bacterial challenge. During mastitis, losses in milk production and compositional changes were most pronounced in infected glands and in severe responders. Milk production of bST cows recovered better than that of placebo cows. Recovery of milk components was accelerated in severe responders treated with bST, but not in moderate responders. Pretreatment of severe responders with bST enhanced milk production before infection, protected the mammary glands from excessive loss of milk during the subsequently induced coliform mastitis, and accelerated normalization of milk composition. In conclusion, the beneficial effects of bST upon normalization of milk production and composition in periparturient cows suffering from coliform mastitis seem to be restricted to the severe responders. In severe responders that had been treated with bST, changes observed during mastitis resembled those in moderate responders treated with the placebo

Molecular cloning and biochemical characterization of a Cu,Zn-superoxide dismutase from Scedosporium apiospermum.

A Cu,Zn-superoxide dismutase has been characterized from Scedosporium apiospermum, a fungus which often colonizes the respiratory tract of patients with cystic fibrosis. Enzyme production was stimulated by iron starvation. Purification was achieved from mycelial extract from 7-day-old cultures on Amberlite XAD-16. The purified enzyme presented a relative molecular mass of 16.4 kDa under reducing conditions and was inhibited by potassium cyanide and diethyldithiocarbamate, which are two known inhibitors of Cu,Zn-SODs. Its optimum pH was 7.0 and the enzyme retained full activity after pretreatment at temperatures up to 50 degrees C. Moreover, a 450-bp fragment of the gene encoding the enzyme was amplified by PCR using degenerate primers designed from sequence alignment of four fungal Cu,Zn-SODs. Sequence data from this fragment allowed us to design primers which were used to amplify by walking-PCR the flanking regions of the known fragment. SaSODC gene (890 bp) corresponded to a 154 amino acid polypeptide with a predicted molecular mass of 15.9 kDa. A database search for sequence homology revealed for the deduced amino acid sequence 72 and 83% identity rate with Cu,Zn-SODs from Aspergillus fumigatus and Neurospora crassa, respectively. To our knowledge, this enzyme is the first putative virulence factor of S. apiospermum to be characterized

An Assessment of Food Safety Handling Practices at Farmers\u27 Markets in Rhode Island Using a Smartphone Application

The number of foodborne illness outbreaks increased in the US from 2001 to 2010, and 17% of them were associated with produce. Higher risk, whole produce sold at farmers\u27 markets presents unique challenges to food safety practices in regard to temperature controls, potable water, and exposure to contaminants. The purpose of this study is to use direct observations to identify unsafe food handling practices among vendors selling higher risk produce at Rhode Island farmers\u27 markets. This study used, as a tool for data acquisition, a Smartphone application developed to allow concealed direct observations of actual vendors\u27 practices at farmers\u27 markets. Observations were made at fourteen (7 state and 7 private) farmers\u27 markets to collect data on food handling practices of 26 vendors selling higher-risk produce. The results of this study will be used as guidance for education programs targeting farmers\u27 market managers and vendors that promote best practices in regard to whole produce

Cell-Free DNA as a Diagnostic and Prognostic Biomarker in Pediatric Rhabdomyosarcoma.

PURPOSE: Total cell-free DNA (cfDNA) and tumor-derived cfDNA (ctDNA) can be used to study tumor-derived genetic aberrations. We analyzed the diagnostic and prognostic potential of cfDNA and ctDNA, obtained from pediatric patients with rhabdomyosarcoma. METHODS: cfDNA was isolated from diagnostic plasma samples from 57 patients enrolled in the EpSSG RMS2005 study. To study the diagnostic potential, shallow whole genome sequencing (shWGS) and cell-free reduced representation bisulphite sequencing (cfRRBS) were performed in a subset of samples and all samples were tested using droplet digital polymerase chain reaction to detect methylated RASSF1A (RASSF1A-M). Correlation with outcome was studied by combining cfDNA RASSF1A-M detection with analysis of our rhabdomyosarcoma-specific RNA panel in paired cellular blood and bone marrow fractions and survival analysis in 56 patients. RESULTS: At diagnosis, ctDNA was detected in 16 of 30 and 24 of 26 patients using shallow whole genome sequencing and cfRRBS, respectively. Furthermore, 21 of 25 samples were correctly classified as embryonal by cfRRBS. RASSF1A-M was detected in 21 of 57 patients. The presence of RASSF1A-M was significantly correlated with poor outcome (the 5-year event-free survival [EFS] rate was 46.2% for 21 RASSF1A-M‒positive patients, compared with 84.9% for 36 RASSF1A-M‒negative patients [P < .001]). RASSF1A-M positivity had the highest prognostic effect among patients with metastatic disease. Patients both negative for RASSF1A-M and the rhabdomyosarcoma-specific RNA panel (28 of 56 patients) had excellent outcome (5-year EFS 92.9%), while double-positive patients (11/56) had poor outcome (5-year EFS 13.6%, P < .001). CONCLUSION: Analyzing ctDNA at diagnosis using various techniques is feasible in pediatric rhabdomyosarcoma and has potential for clinical use. Measuring RASSF1A-M in plasma at initial diagnosis correlated significantly with outcome, particularly when combined with paired analysis of blood and bone marrow using a rhabdomyosarcoma-specific RNA panel