Spatial coupling of mTOR and autophagy augments secretory phenotypes

Protein synthesis and autophagic degradation are regulated in an opposite manner by mammalian target of rapamycin (mTOR), whereas under certain conditions it would be beneficial if they occurred in unison to handle rapid protein turnover. We observed a distinct cellular compartment at the trans side of the Golgi apparatus, the TOR-autophagy spatial coupling compartment (TASCC), where (auto)lysosomes and mTOR accumulated during Ras-induced senescence. mTOR recruitment to the TASCC was amino acid- and Rag guanosine triphosphatase-dependent, and disruption of mTOR localization to the TASCC suppressed interleukin-6/8 synthesis. TASCC formation was observed during macrophage differentiation and in glomerular podocytes; both displayed increased protein secretion. The spatial coupling of cells' catabolic and anabolic machinery could augment their respective functions and facilitate the mass synthesis of secretory proteins.     

Biological Potential of Flaxseed Protein Hydrolysates Obtained by Different Proteases

Plant Foods for Human Nutrition - Flaxseed meal, a byproduct of flaxseed oil extraction, was treated as low-value agrowaste for a long time despite its high protein content. Flaxseed meal has...     

Pseudocereals as alternative sources for high folate content in staple foods

Total folate content was determined in the pseudocereals amaranth (four varieties), quinoa and buckwheat in comparison to four cereal species (eight wheat varieties, four barley varieties, one oat variety, one rye variety). Amaranth and quinoa were found to possess very high total folate contents: in amaranth, total folate ranged from 52.8 to 73.0 and in quinoa it was 132.7 μg/100 g dm, about ten times as much as in wheat. The bran fractions contained on average 124% of total folate, while only 57% on average was present in the flour fractions.     

Characterisation of gluten-free pasta through conventional and innovative methods: Evaluation of the cooking behaviour

The rapid stiffening of a well-developed gluten network able to entrap swollen starch granules is a key factor for the high quality of durum wheat pasta during cooking. Good resistance and firmness, low stickiness and limited release of organic materials into the cooking water are quality traits of primary importance for traditional pasta. In gluten-free (GF) pasta, the formation of a scaffold of retrograded starch can be an alternative to gluten networking: it confers rigidity to the cooked product and reduces pasta stickiness and loss of soluble materials into the cooking water. In a previous paper, 14 commercial GF spaghetti samples were studied as uncooked products from a chemical, biochemical and physical point of view. The aim of this study was to determine the cooking behaviour of these samples. A durum wheat pasta was also included as reference. Suitable cooking conditions were adopted and different conventional and innovative evaluations (i.e. compression test, creep test) were performed as a function of cooking time. Different behaviours were evidenced, often related to the ultrastructural organization of the uncooked products. In particular, the creep test revealed to be very effective in discriminating among the properties of the different GF spaghetti.     

Towards the identification of ecological management units: A multidisciplinary approach for the effective management of bottlenose dolphins in the southern Iberian Peninsula

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Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs

Because infected dogs are widely considered to be the main domestic reservoir for Leishmania infantum (syn Leishmania chagasi) parasites in Brazil, the diagnosis of canine visceral leishmaniasis (CVL) must be made both accurately and promptly. The present study attempted to standardize a conventional polymerase chain reaction (cPCR) protocol for the detection of L. infantum DNA in canine spleen samples. Quantitative PCR (qPCR) technique was used to confirm the presence of Leishmania DNA in the canine spleen fragments. A comparison was made between the efficacies of these molecular diagnostic techniques and conventional parasitological and serological methods. cPCR protocols for spleen samples were standardized using primers that amplify a 145 bp fragment, located at the parasite kinetoplast minicircle. The genus specificity of the cPCR protocol was assessed by its inability to amplify the DNA of other common canine pathogens, such as Ehrlichia canis, Babesia canis, Toxoplasma gondii and Trypanosoma cruzi. cPCR protocol sensitivity was tested by assessing the reaction detection limit, determined to be 10 fg of L. infantum reference strain DNA, which corresponds to a range of 0.03-0.1 parasites per fragment. Standardized cPCR protocol was used to detect the presence of Leishmania in 45 dog spleen samples. Our results showed that 40% of the spleen fragment cultures were positive for Leishmania parasites, 58% of the dog serum samples tested positive using ELISA, and parasite DNA was detected in 44% using qPCR, while 47% of the spleen samples using cPCR. Diagnostic methods performance was assessed and revealed a better degree of ascertainment for cPCR when compared to other diagnostic methods. The sensitivity of ELISA was 83.3%, qPCR was 83.3%, and cPCR was 88.9%; PPV for ELISA was 57.7%, qPCR was 75% and cPCR was 76.2%; the Kappa coefficients were found to be 0.40 (fair) for ELISA, 0.64 (substantial) for qPCR and 0.68 (substantial) for cPCR. In both oligosymptomatic and polysymptomatic dogs, cPCR revealed the better performance analysis when compared to other diagnostic methods. The findings presented herein establish cPCR as the most indicated test to detect Leishmania when compared to the other two diagnostic methods evaluated. Despite the fact that the qPCR protocol provides a highly accurate quantification of parasites when targeting the SSU rRNA gene, this technique does not significantly improve the diagnosis of CVL when compared with the performance of the cPCR protocol, which focused on the kinetoplast minicircle.     

Selection for tetracycline-resistant Chlamydia suis in treated pigs

The aim of this study was to investigate Chlamydia suis in a pig farm with an outbreak of conjunctivitis and diarrhea. Eye swabs and pooled fecal samples were investigated for the presence of C. suis by real-time PCR and ArrayTube microarray. Samples positive for C. suis by ArrayTube microarray assay were further tested for the presence of the tet(C) resistance gene by PCR. In the first examination, C. suis was identified in 12 six-week-old pigs showing conjunctivitis. Of these, the tet(C) gene-coding region was amplified in one pooled fecal sample and one eye swab, respectively. After oral treatment with tetracycline, clinical symptoms disappeared. Subsequently, all eye swabs investigated from 10 healthy pigs were positive for C. suis and the tet(C) gene-coding region. The present study reports rapid selection for tetracycline-resistant C. suis after antibiotic treatment.     

Evaluation of a minimally invasive non-calibrated pulse contour cardiac output monitor (FloTrac/Vigileo) in anaesthetized dogs

ObjectiveTo evaluate the accuracy of a new cardiac output monitor (FloTrac/Vigileo), originally designed for humans, in dogs. This pulse contour cardiac output monitoring system cannot be calibrated and measures cardiac output (

t) from a standard arterial catheter.Study designProspective experimental trial.AnimalsEight adult Beagle dogs weighing 13.1 (9.8–17.1) kg [median (range)].MethodsAnaesthesia in the dogs was maintained using isoflurane. A pulmonary artery catheter and a metatarsal arterial catheter (22 gauge) were placed. Cardiac output was measured simultaneously 331 times by thermodilution and FloTrac technique. A broad spectrum of

t measurements was achieved through alterations of isoflurane concentration, administration of propofol boluses and dobutamine infusions. Agreement between the methods was quantified with Bland Altman analysis and disagreement was assessed with linear mixed models.ResultsMedian (10th and 90th percentile) cardiac output as measured with thermodilution was 2.54 (1.47 and 5.15) L minute^?1 and as measured with FloTrac 8.6 (3.9 and 17.3) L minute^?1. FloTrac measurements were consistently higher with a mean bias of 7 L minute^?1 and limits of agreement of ?3.15 to 17.17 L minute^?1. Difference between the methods was most pronounced in high

t measurements. Linear mixed models showed an estimated difference between the two methods of 8.05 (standard error 1.18) L minute^?1 and a significant interaction between mean arterial pressure and method. Standard deviation (4.45 higher) with the FloTrac method compared to thermodilution was increased.ConclusionCompared to thermodilution measurements, the FloTrac system was influenced to a higher degree by arterial blood pressure, resulting in consistent overestimation of cardiac output.Clinical RelevanceThe FloTrac monitor, whose algorithms were developed based on human data, cannot be used as an alternative for thermodilution in dogs.     

Risk factors for highly pathogenic H7N1 avian influenza virus infection in poultry during the 1999-2000 epidemic in Italy

In 1999-2000, Italian poultry production was disrupted by an H7N1 virus subtype epidemic of highly pathogenic avian influenza (HPAI). The objectives of the present study were to identify risk factors for infection on poultry farms located in regions that had the highest number of outbreaks (Veneto and Lombardia) and the impact of pre-emptive culling as a complementary measure for eradicating infection. A Cox regression model that included spatial factors, such as the G index, was used. The results confirmed the relationship between risk of infection and poultry species, production type and size of farms. The effectiveness of pre-emptive culling was confirmed. An increased risk of infection was observed for poultry farms located near an infected farm and those at altitudes less than 150m above sea level. The measures for the control and eradication of AI virus infection need to consider species differences in susceptibility, the types of production and the density of poultry farms in the affected areas.