Biofilm formation of Staphylococcus aureus dairy isolates representing different genotypes.

The objective of this study was to compare the biofilm-forming capabilities of different genotypes of Staphylococcus aureus dairy isolates from Switzerland and northern Italy, including Staph. aureus genotype B (GTB) and methicillin-resistant Staph.aureus (MRSA). We hypothesized that biofilm formation might be more pronounced in the contagious GTB isolates compared with other genotypes affecting individual animals. Twenty-four dairy isolates, including 9 MRSA, were further characterized by genotyping by using ribosomal spacer PCR, spa typing, biofilm formation under static and dynamic conditions, and scanning electron microscopy. The GTB isolates (n = 6) were more able to form biofilms than other genotypes at 37°C and at 20°C after 48 and 72 h of incubation in the static assay using polystyrene microtiter plates. This result was supported by scanning electron micrographs showing a GTB isolate producing strong biofilm with extracellular matrix in contrast to a genotype C isolate. Furthermore, none of the MRSA isolates formed strong biofilms in the static assay. However, some MRSA produced low or moderate amounts of biofilm depending on the applied conditions. Under dynamic conditions, a much more diverse situation was observed. The ability of GTB isolates to be strong biofilm formers was not observed in all cases, emphasizing the importance of growth conditions for the expression of biofilm-related genes. No specific genotype, spa type, or MRSA isolate could be categorized significantly into one level of biofilm formation. Nineteen percent of isolates behaved similarly under static and dynamic conditions. The results of this study expand our knowledge of different dairy-related Staph. aureus subtypes and indicate the benefit of genotyping when biofilms are studied.

Advanced soft-tissue sarcoma in elderly patients: patterns of care and survival.

BACKGROUND: There are no data regarding the management of advanced soft-tissue sarcoma (STS) in elderly patients. PATIENTS AND METHODS: We retrospectively reviewed the charts of patients ≥75 years old diagnosed with metastatic or unresectable STS between 1991 and 2011 in 11 French and American centers. RESULTS: The study included 361 patients. Of these, 223 patients (62%) received systemic therapy, whereas 123 patients (34%) were managed with best supportive care (BSC) only. Patients who received BSC were more likely to be ≥80 years, with performance status (PS) ≥ 2, Charlson comorbidity score ≥ 10, and metastatic disease. The median progression-free survival of patients treated with systemic therapy was 4 months (95% CI: 2.9-5.1). Thirty-six patients (16%) stopped chemotherapy because of toxicity. Median overall survival (OS) of patients managed with specific therapy was 10.9 months (95% CI: 8.3-13.5) versus 5.3 months (95% CI: 3.6-7.1) for patients managed with BSC (P = 0.001). On multivariate analysis, age ≥ 80 years, PS ≥ 2, and number of metastatic sites were the only independent factors associated with OS. CONCLUSION: A high proportion of elderly patients with advanced STS were denied chemotherapy. Further efforts are needed to define better the optimal care for fit and unfit elderly patients with STS.

An Italian multicenter retrospective real-life analysis of patients with brain metastases from renal cell carcinoma: the BMRCC study.

BACKGROUND: The treatment of patients with brain-spread renal cell carcinoma (RCC) is an unmet clinical need, although more recent therapeutic strategies have significantly improved RCC patients' life expectancy. Our multicenter, retrospective, observational study investigated a real-world cohort of patients with brain metastases (BM) from RCC (BMRCC). PATIENTS AND METHODS: A total of 226 patients with histological diagnosis of RCC and radiological evidence of BM from 22 Italian institutions were enrolled. Univariate and multivariate models were performed to investigate the impact of clinicopathological features and multimodal treatments on both overall survival (OS) from the BM diagnosis and intracranial progression-free survival (iPFS). RESULTS: The median OS from the BM diagnosis was 18.8 months (interquartile range: 6.2-43 months). Multivariate analysis confirmed the following as positive independent prognostic factors: a Karnofsky Performance Status >70% [hazard ratio (HR) = 0.49, 95% confidence interval (CI) 0.26-0.92, P = 0.0026] and a single BM (HR = 0.51, 95% CI 0.31-0.86, P = 0. 0310); in contrast, the following were confirmed as worse prognosis factors: progressive extracranial disease (HR = 1.66, 95% CI 1.003-2.74, P = 0.00181) and only one line of systemic therapy after the BM occurrence (HR = 2.98, 95% CI 1.62-5.49, P = 0.029). Subgroup analyses showed no difference in iPFS according to the type of the first systemic treatment [immunotherapy (IT) or targeted therapy (TT)] carried out after the BM diagnosis (HR = 1.033, 95% CI 0.565-1.889, P = 0.16), and revealed that external radiation therapy (eRT) significantly prolonged iPFS when combined with IT (10.7 months, 95% CI 4.9-48 months, P = 0.0321) and not when combined with TT (9.01 months, 95% CI 2.7-21.2 months, P = 0.59). CONCLUSIONS: Our results suggest a potential additive effect in terms of iPFS for eRT combined with IT and encourage a more intensive multimodal therapeutic strategy in a multidisciplinary context to improve the survival of BMRCC patients.

Antimicrobial resistance pattern of methicillin-resistant Staphylococcus aureus in the food industry.

There is increasing concern about the impact on public health of methicillin-resistant Staphylococcus aureus (MRSA) associated with animal food products. MRSA remains a serious problem because of the high incidence and multidrug resistance of the strains, even for strains isolated from foods, food environments and food handlers. The objectives of this study are: (i) to evaluate the susceptibility of S. aureus strains isolated from food, food handlers and food-processing environments to 14 antibiotics currently used in veterinary and human therapy; (ii) to assess the presence of the mecA gene. A total of 1007 samples were collected from food, food handlers, and environments and were analyzed for the presence of S. aureus. S. aureus was present in 165 of the 1007 samples. A total of 157 isolates were methicillin-susceptible S. aureus (MSSA) and 8 isolates were MRSA. In particular, out of 8 MRSA strains detected, 4 strains harboured the mecA gene. All MRSA strains were resistant to at least one of the tested antibiotics and 6 strains demonstrated multi-resistance. Considering the high level of resistances in S. aureus and the isolation of MRSA strains, the surveillance of antimicrobial resistance and the spreading of this pathogen is of crucial importance in the food production chain. These data are useful in improving background data on antimicrobial resistance of S. aureus isolated from food, processing environments and food handlers, supporting the prudent use of antibiotics and the development of international control programs.

Distribution of aminogenic activity among potential autochthonous starter cultures for dry fermented sausages.

Any bacterial strain to be used as starter culture should have suitable characteristics, including a lack of amino acid decarboxylase activity. In this study, the decarboxylase activity of 76 bacterial strains, including lactic acid bacteria and gram-positive, catalase-positive cocci, was investigated. These strains were previously isolated from European traditional fermented sausages to develop autochthonous starter cultures. Of all the strains tested, 48% of the lactic acid bacteria strains and 13% of gram-positive, catalase-positive cocci decarboxylated one or more amino acids. Aminogenic potential was strain dependent, although some species had a higher proportion of aminogenic strains than did others. Thus, all Lactobacillus curvatus strains and 70% of Lactobacillus brevis strains had the capacity to produce tyramine and beta-phenylethylamine. Some strains also produced other aromatic amines, such as tryptamine and the diamines putrescine and cadaverine. All the enterococcal strains tested were decarboxylase positive, producing high amounts of tyramine and considerable amounts of beta-phenylethylamine. None of the staphylococcal strains had tyrosine-decarboxylase activity, but some produced other amines. From the aminogenic point of view, Lactobacillus plantarum, Lactobacillus sakei, and Staphylococcus xylosus strains would be the most suitable for use as autochthonous starter cultures for traditional fermented sausages.

Traditional dry fermented sausages produced in small-scale processing units in Mediterranean countries and Slovakia. 1: Microbial ecosystems of processing environments.

Microbial ecosystems were surveyed in 314 environmental samples from 54 Southern and Eastern European small-scale processing units (PUs) manufacturing traditional dry fermented sausages. The residual microflora contaminating the surfaces and the equipment were analysed after cleaning and disinfection procedures. All the PU environments were colonised at various levels by spoilage and technological microflora with excessive contamination levels in some of the PUs. Sporadic contamination by pathogenic microflora was recorded. Salmonella and Listeria monocytogenes were detected in 4.8% and 6.7% of the samples, respectively, and Staphylococcus aureus was enumerated in 6.1% of the samples. Several critical points were identified, such as the machines for S. aureus and the tables and the knives for L. monocytogenes; this knowledge is crucial for the improvement of hygiene control systems in small and traditional meat processing industries. The variability of the residual contamination emphasized the different cleaning, disinfecting and manufacturing practices routinely followed by these small-scale processing units.

Metal Concentrations in Samples of Frozen Cephalopods (Cuttlefish, Octopus, Squid, and Shortfin Squid): An Evaluation of Dietary Intake.

Cephalopods are an important source of nutrients and some of the most widely consumed marine foods. However, because of contamination of the oceans and the bioaccumulative nature of toxic metals, these foods may pose a health risk. For this reason, the concentrations of some trace elements (chromium [Cr], lithium, strontium [Sr], copper [Cu], and nickel) and toxic metals (aluminum [Al], cadmium, and lead) were determined in 65 frozen samples of cuttlefish, octopus, common squid, and shortfin squid by inductively coupled plasma optical emission spectrometry to evaluate dietary intake and toxic risk. Sr was the major trace element (3.03 mg/kg) in cuttlefish; however, Cu (1.57 mg/kg) was found in the highest concentration in common squid. Among the toxic metals, Al had the highest concentration (3.09 mg/kg) in common squid. Al can pose an important health risk to individuals with kidney problems and to children because these groups are most vulnerable to the toxic effects. Significant differences among the four cephalopod types were found in the concentrations of most of the metals examined. Taking into account the average consumption of cephalopods, the contribution of toxic metals does not pose a risk to the health of adults.

Targeting androgen-independent pathways: new chances for patients with prostate cancer?

Androgen deprivation therapy (ADT) is the mainstay treatment for advanced prostate cancer (PC). Most patients eventually progress to a condition known as castration-resistant prostate cancer (CRPC), characterized by lack of response to ADT. Although new androgen receptor signaling (ARS) inhibitors and chemotherapeutic agents have been introduced to overcome resistance to ADT, many patients progress because of primary or acquired resistance to these agents. This comprehensive review aims at exploring the mechanisms of resistance and progression of PC, with specific focus on alterations which lead to the activation of androgen receptor (AR)-independent pathways of survival. Our work integrates available clinical and preclinical data on agents which target these pathways, assessing their potential clinical implication in specific settings of patients. Given the rising interest of the scientific community in cancer immunotherapy strategies, further attention is dedicated to the role of immune evasion in PC.

Performances and meat quality of two Italian pig breeds fed diets for commercial hybrids.

The effects of specific diets for commercial hybrids were investigated on 6 Casertana and 11 Mora Romagnola, two endangered Italian pig breeds. Average daily gain (ADG), feed conversion index (FCI), dressing percentage and meat and fat quality of animals bred under similar environmental and nutritional conditions were compared to define their optimal slaughtering weight. Animals were fed the same diets assuming that requirements of Mora Romagnola and Casertana did not differ, and changed every 30kg of weight gain. ADG and FCI were calculated every 15 days. Weight gains, divided into 5 groups based on live weight (LW) of animals (⩽60kg, 60160kg), showed higher values for Casertana than Mora Romagnola, particularly from 121 to 160kg LW (687g/d). Average FCI from 50 to 160kg LW was similar in both breeds (4.2). After 403 days of trial, animals were slaughtered at about 195kg LW. Carcass measurements showed that Casertana had higher dressing percentage and lean cuts than Mora Romagnola. Both breeds showed extraordinary high ultimate pH values of M. longissimus thoracis (5.96 and 6.15 for Casertana and Mora Romagnola, respectively) M. semimembranosus (6.37 and 6.30), showing an incomplete post mortem glycolysis. Colour of M. longissimus thoracis did not differ between breeds and was particularly dark. Chemical analysis of Casertana meat showed lower percentage of water and fat; the total amount of fatty acids (SFA, MUFA and PUFA) and the SFA/UFA ratio did not show significant differences between breeds. Results showed that from a growth point of view the optimal slaughtering weight of Casertana and Mora Romagnola should not exceed 160kg LW. Both breeds had an uncommon reactivity to stress probably due to interactions of genetic, nutritional and management factors.

Tibolone: a review.

Tibolone appears to be at least as efficacious as other forms of hormonal replacement therapy (HRT) on climacteric symptoms. It does not cause withdrawal bleeding when used in women with at least 1 year of amenorrhea. It is, therefore, not indicated in perimenopause because it may cause irregular bleeding. The androgenic action of tibolone may have a two-fold benefit: on the one hand, it may help depression and libido more than other forms of HRT, while, on the other hand, it may improve some lipid parameters such as Lp(a), and triglycerides. However, this androgenic action, may also be responsible for the reduction of HDL cholesterol, that may thus reduce the beneficial effect of tibolone on lipids. It is estimated that only 30% of cardiovascular risk protection of HRT is due to improvement of classical lipids parameters while a great role is played by the direct effect of estrogen on vessels. Tibolone, as well as estrogen, has been shown to induce peripheral vasodilatation and also has a direct effect on vascular reactivity thus increasing peripheral blood flow with no changes in blood pressure or cardiac output. Tibolone seems to exert a similar effect as other forms of HRT on markers of bone metabolism and bone mass, but no data is yet available on fracture prevention.

Oxidation in traditional mediterranean meat products.

Lipid oxidation in dry-fermented sausages and dry-salted-hams, representative items of traditional Mediterranean pork products, is responsible for the production of compounds which can affect their organoleptic qualities and wholesomeness. Lipid-derived compounds make up an important share of the volatiles extracted from matured products, once the contribution of spices is removed, and contribute significantly to the flavour of old style products. The few reports available on oxysterols depict a total cholesterol oxidation rate of about 0.1%, a threshold level for toxic effects to occurin vitro but about 100 times lower of thein vivo toxicity dose. The value could perhaps be reduced with the use of antioxidants. Phospholipids are the main class of fats interested by lipolysis and unsaturated fatty acids are those most interested by oxidation. On that basis, the case of special feeding regimes, such as that of Iberian pigs, and the possible relevance of muscle fibre type on flavour development are discussed.

Lipid and colour stability of Milano-type sausages: effect of packing conditions.

Lipid and colour oxidative changes in Milano-type fermented sausages were studied in relation to packing conditions and extended storage under fluorescent light. Matured sausages were sliced and packed under vacuum or in protective atmosphere (100% N(2)) and exposed in a display cabinet to mimic commercial conditions of light and temperature for 2 months. Lipid oxidation was measured by the determination of thiobarbituric acid reactive substances (TBARS) and cholesterol oxidation products (COP), whereas the oxidation of nitrosylmyoglobin was evaluated by a trained sensory panel. Lipid oxidation and discolouration were positively related in vacuum-packed sausages: TBARS and COP values increased significantly concurrently with increasing brown scores. Protective atmosphere came out to be more efficient than vacuum in controlling fatty acid oxidation and, to a lesser extent, cholesterol and pigment degradation: TBARS values remained constant during the whole storage period whereas cholesterol oxides and brown colour scores gradually increased but remained lower than those of vacuum-packed sausages. Higher residual oxygen in vacuum packing could be responsible for the observed differences in oxidative stability.

Volatile compounds of commercial Milano salami.

The relationship between extracted volatiles of Milano salami, one of the main dry-cured sausages produced in Italy, and their olfactory properties was studied. Volatile compounds were extracted by a purge-and-trap method, quantified using a flame ionisation detector and identified by mass spectrometry. Olfactory analysis was performed by sniffing the gas chromatographic effluent. Nearly 80 compounds were identified and quantified: most came from spices (60.5%), 18.9% from lipid oxidation, 11.8% from amino acid catabolism and 4.9% from fermentation processes. Panellists detected 19 odours by sniffing. These odours were associated with spices, lipid oxidation or fermentation and were in agreement with the contributions of each reaction to the overall aroma of the product.

Oxidative stability of lipids and cholesterol in salame Milano, coppa and Parma ham: dietary supplementation with vitamin E and oleic acid.

An investigation has been carried out on the effect of dietary supplementation with vitamin E and oleic acid on oxidative stability of Italian representative pork products. Fatty acid composition of deposited fat was modified in the oil supplemented groups with negative effects on fat firmness. Oil supplementation significantly increased vitamin E content of muscles, as well. Oxidative stability of fat, evaluated by TBARS and cholesterol oxides, has not shown significant differences between dietary treatments but, although the differences were not significant, the tendency was sufficiently clear, especially in salame Milano, towards a lower oxidation level in vitamin E enriched meat. Cholesterol oxidation was found to vary in general around 0.1% of total cholesterol and the only cholesterol oxides observed were 7ß-hydroxycholesterol, 5,6α-epoxycholesterol and 7-ketocholesterol. Aldehydes content and distribution appeared to be linked, firstly, to the specific processing technology of the product and, secondly, to the changed fatty acid composition.

Volatile compounds released during ripening in Italian dried sausage.

A commercial production was analysed at six stages during ripening. Water content, pH and bacterial counts were followed, and volatile compounds from sausages were extracted by dynamic headspace sampling and analysed by gas chromatography/mass spectrometry. Total concentrations of all classes increased during ripening. Pepper compound concentrations peaked in the middle of the ripening period. Lipid oxidation products increased especially towards the end of ripening, in particular, the compounds 2-heptanol, 1-octen-3-ol, 2-heptanone and 2-nonanone. Surface moulds probably caused 4-heptanone to appear late in the processing. Benzeneacetaldehyde was absent in fresh mince, but increased to become one of the most abundant volatiles. Compounds from carbohydrate catabolism disappeared during the processing.

Lipid and cholesterol oxidation in frozen stored pork, salame Milano and mortadella.

An investigation has been carried out on oxidative processes of fresh and stored frozen pork and of two pork products (salame Milano and mortadella). Wholesale pork cuts (shoulder, ham trimmings, belly and backfat) were checked fresh and after frozen storage (1, 3 and 6 months). The streaky bacon was studied fresh and after scalding at 80 °C. Salame Milano and mortadella were produced with fresh and frozen raw materials and a few commercial samples were also part of the study. No real differences have been observed between commercial and experimentally produced salame and mortadella. Peroxide and TBARS values were low in all cases. Among the cholesterol oxides determined (7ß-hydroxycholesterol, 5,6α-epoxycholesterol, 7-ketocholesterol and 25-hydroxycholesterol) those dangerous for human health were either absent or their concentration was below toxic levels. The results are discussed on the light of literature reports and Italian pork products technologies.

Comparative evaluation of some antioxidants in salame Milano and mortadella production.

Six formulations of salame Milano and mortadella were produced, one without antioxidants, two with sodium ascorbate (0.05-0.1 and O.2%), one with catechin, one with phytic acid, and one with sesamol. The products were studied for oxidation levels (peroxide value, TBARS, cholesterol oxides) and colour stability. The evaluation of the latter took place in conditions simulating commercial types of packaging (vacuum and modified atmosphere) and display. A significant difference emerged in colour stability. With the exception of salame Milano packed under modified atmosphere, in all cases salame and mortadella produced without Na ascorbate had significantly lower colour stability. Both in salame and mortadella, peroxide values and TBARS determination gave indications of a possible pro-oxidant effect of ascorbate 0.2% compared with an effective antioxidant activity of the other compounds tested. Cholesterol oxidation in salame appeared to be inhibited by sesamol and ascorbate while catechin was much less effective. In mortadella the prooxidant effect of Na ascorbate was confirmed. All lipid oxidation parameters, though, showed low oxidation values so that no quality nor health deficiencies could be envisaged with any of the formulations adopted. Lower colour stability scores, though, would create commercial problems.

Lean colour of green and matured Parma hams: Comparative evaluation and technological relevance of sensory and objective data.

Research has been conducted on the relationship between sensory evaluation and objective measurement of pork colour. The study also investigated the relationship between the colour of fresh meat and the colour of matured hams. Sensory evaluation was carried out at 24 h post mortem on the freshly cut surfaces of green hams trimmed for Parma ham production. Objective measurements were performed at the same time on Semimembranosus and Biceps femoris muscles with a Minolta Chromameter Reflectance II CR200/08. Cold carcass weight, lean content, trimmed ham weight, 45 min and 24 h post mortem pH of Semimembranosus muscle were also recorded. The hams were processed according to Parma ham technology. Weight losses at standard processing times were collected. At the end of processing a sample of about one-third of the hams was evaluated for internal lean colour and lean firmness both by sensory panel and objective measurements. Proximate composition was also determined. The results showed highly significant relationships between colour sensory scores and the coordinates L (∗), b (∗) and hue of the C.I.E. L (∗)a (∗)b (∗) colour system. Among the other parameters measured on fresh meat only 24 h pH was related at a very significant level with sensory and objective colour values. Correlation coefficients were generally higher for Semimembranosus than Biceps femoris. Matured ham colour was linked with fresh meat sensory judgements, L (∗), hue and a (∗) values. The relationship, though, was not sufficiently strong for use in predicting the colour of matured hams from green ham characteristics. Processing appears to have a positive effect on the colour of pale meat. Weight losses were mainly linked with lean meat content; other quality attributes being less important. Lean firmness of matured hams was related to the colour and pH of the fresh meat.

Oxidative stability and dietary treatment with vitamin E, oleic acid and copper of fresh and cooked pork chops.

Research has been conducted on the effects of feed supplementation with vitamin E, oleic acid (sunflower oil) and copper on some quality characteristics of pork chops. The study was based on the measurement of pH 45' and 24 hr post mortem, proximate composition, colour both objective and sensory, colour stability after packing in oxygen permeable film and protective atmosphere, copper and iron contents, vitamin E content in fresh and cooked chops, peroxide and TBARS value in raw and cooked meat, total cholesterol, cholesterol oxides and aldehydes in cooked meat, sensory evaluation of the eating quality of cooked chops and iodine number of subcutaneous fat. Vitamin E content was significantly higher with oil and vitamin supplementation. No variation has been observed in copper and iron contents. A higher stability of the colour of meat packed under modified atmosphere has been observed with increasing vitamin E content, but the same did not apply with meat packed in oxygen permeable film. No differences have been observed on fatty acid oxidative stability of fresh m. longissimus dorsi and cooked chops (peroxide and TBARS values), on cholesterol oxidation and aldehydes production of cooked chops with increasing vitamin E content. Iodine number in supplemented animals reached levels normally considered at risk for fat firmness.