TP53 mutations and survival in ovarian carcinoma patients receiving first-line chemotherapy plus bevacizumab: Results of the MITO16A/MaNGO OV-2 study.

To date, there are no biomarkers that define a patient subpopulation responsive to bevacizumab (BEV), an effective treatment option for advanced ovarian carcinoma (OC). In the context of the MITO16A/MaNGO OV-2 trial, a Phase IV study of chemotherapy combined with BEV in first-line treatment of advanced OC, we evaluated TP53 mutations by next-generation sequencing and p53 expression by immunohistochemistry (IHC) on 202 and 311 cases, respectively. We further correlated TP53 mutations in terms of type, function, and site, and IHC data with patients' clinicopathological characteristics and survival. TP53 missense mutations of unknown function (named unclassified) represented the majority of variants in our population (44.4%) and were associated with a significantly improved overall survival (OS) both in univariable (hazard ratio [HR] = 0.43, 95% confidence interval [CI] = 0.20-0.92, p = .03) and multivariable analysis (HR = 0.39, 95% CI = 0.18-0.86, p = .02). Concordance between TP53 mutational analysis and IHC was 91%. We observed an HR of 0.70 for OS in patients with p53 IHC overexpression compared to p53 wild-type, which however did not reach statistical significance (p = .31, 95% CI = 0.36-1.38). Our results indicate that the presence of unclassified TP53 mutations has favorable prognostic significance in patients with OC receiving upfront BEV plus chemotherapy. In particular, unclassified missense TP53 mutations characterize a subpopulation of patients with a significant survival advantage, independently of clinicopathological characteristics. Our findings warrant future investigations to confirm the prognostic impact of TP53 mutations in BEV-treated OC patients and deserve to be assessed for their potential predictive role in future randomized clinical studies.

The Bee Gut Microbiota: Bridging Infective Agents Potential in the One Health Context.

The bee gut microbiota plays an important role in the services the bees pay to the environment, humans and animals. Alongside, gut-associated microorganisms are vehiculated between apparently remote habitats, promoting microbial heterogeneity of the visited microcosms and the transfer of the microbial genetic elements. To date, no metaproteomics studies dealing with the functional bee microbiota are available. Here, we employ a metaproteomics approach to explore a fraction of the bacterial, fungal, and unicellular parasites inhabiting the bee gut. The bacterial community portrays a dynamic composition, accounting for specimens of human and animal concern. Their functional features highlight the vehiculation of virulence and antimicrobial resistance traits. The fungal and unicellular parasite fractions include environment- and animal-related specimens, whose metabolic activities support the spatial spreading of functional features. Host proteome depicts the major bee physiological activities, supporting the metaproteomics strategy for the simultaneous study of multiple microbial specimens and their host-crosstalks. Altogether, the present study provides a better definition of the structure and function of the bee gut microbiota, highlighting its impact in a variety of strategies aimed at improving/overcoming several current hot topic issues such as antimicrobial resistance, environmental pollution and the promotion of environmental health.

Carboplatin and paclitaxel plus avelumab compared with carboplatin and paclitaxel in advanced or recurrent endometrial cancer (MITO END-3): a multicentre, open-label, randomised, controlled, phase 2 trial.

BACKGROUND: Adding immunotherapy to first-line chemotherapy might improve outcomes for patients with advanced or recurrent endometrial cancer. We aimed to compare carboplatin and paclitaxel versus avelumab plus carboplatin and paclitaxel as first-line treatment with avelumab given concurrent to chemotherapy and as maintenance after the end of chemotherapy. METHODS: MITO END-3 is an open-label, randomised, controlled, phase 2 trial conducted at 31 cancer institutes, hospitals, and universities in Italy. Eligible patients were aged 18 years or older with histologically confirmed advanced (FIGO stage III-IV) or recurrent endometrial cancer, an Eastern Cooperative Oncology Group (ECOG) performance status of 0-1, and no previous systemic anticancer therapy as primary treatment for advanced or metastatic disease. Participants were randomly assigned (1:1) using a computerised minimisation procedure stratified by centre, histology, and stage at study entry, to either receive carboplatin (area under the curve [AUC] 5 mg/mL × min) and paclitaxel (175 mg/m2; standard group) intravenously every 3 weeks for six to eight cycles or avelumab (10 mg/kg intravenously) added to carboplatin and paclitaxel (experimental group) every 3 weeks and then every 2 weeks as a single maintenance treatment after the end of chemotherapy until disease progression or unacceptable toxicity. Patients, treating clinicians, and those assessing radiological examinations were not masked to study treatment. The primary endpoint was investigator-assessed progression-free survival, measured in the intention-to-treat (ITT) population. Patients who received at least one dose of study drug were included in the safety analysis. Experimental group superiority was tested with 80% power and one-tailed α 0·20. This trial is registered with ClinicalTrials.gov (NCT03503786) and EudraCT (2016-004403-31). FINDINGS: From April 9, 2018, to May 13, 2021, 166 women were assessed for eligibility and 39 were excluded. 125 eligible patients were randomly assigned to receive carboplatin and paclitaxel (n=62) or avelumab plus carboplatin and paclitaxel (n=63) and included in the ITT population. The median follow-up was 23·3 months (IQR 13·2-29·6) and was similar between the two groups. 91 progression-free survival events were reported, with 49 events in 62 patients in the standard group and 42 events in 63 patients in the experimental group. The median progression-free survival was 9·9 months (95% CI 6·7-12·1) in the standard group and 9·6 months (7·2-17·7) in the experimental group (HR of progression or death 0·78 [60% CI 0·65-0·93]; one-tailed p=0·085). Serious adverse events were reported more frequently in the experimental group (24 vs seven events in the standard group); neutrophil count decrease was the most frequent grade 3-4 adverse event (19 [31%] of 61 patients in the experimental group vs 26 [43%] of 61 patients in the standard group). Two deaths occurred in the experimental group during treatment (one respiratory failure following severe myositis [possibly related to treatment] and one cardiac arrest [not related to treatment]). INTERPRETATION: Adding avelumab to first-line chemotherapy deserves further testing in patients with advanced or recurrent endometrial cancer, although consideration of mismatch repair status is warranted. FUNDING: Pfizer.

Clinical characteristics and molecular aspects of low-grade serous ovarian and peritoneal cancer: a multicenter, observational, retrospective analysis of MITO Group (MITO 22).

BACKGROUND: Low-grade serous ovarian and peritoneal cancer (LGSC) is a rare disease and few data on the clinical and genomic landscape have been published. METHODS: A retrospective analysis of patients diagnosed with LGSC between 1996 and 2019 was conducted in MITO centers. Objective Response Rate (ORR) to treatments, progression-free survival (PFS) and overall survival (OS) were assessed. Additionally, the tumor molecular profile of 56 patients was evaluated using the Next Generation Sequencing (NGS) FoundationOne CDX (Foundation Medicine®). RESULTS: A total of 128 patients with complete clinical data and pathologically confirmed diagnosis of LGSC were identified. ORR to first and subsequent therapies were 23.7% and 33.7%, respectively. PFS was 43.9 months (95% CI:32.4-53.1) and OS was 105.4 months (95% CI: 82.7-not reached). The most common gene alterations were: KRAS (n = 12, 21%), CDKN2A/B (n = 11, 20%), NRAS (n = 8, 14%), FANCA (n = 8, 14%), NF1 (n = 7, 13%) and BRAF (n = 6, 11%). Unexpectedly, pathogenetic BRCA1 (n = 2, 4%), BRCA2 (n = 1, 2%) and PALB2 (n = 1, 2%) mutations were found. CONCLUSIONS: MITO 22 suggests that LGSC is an heterogenous disease for both its clinical behavior in response to standard therapies and its molecular alterations. Future prospective studies should test treatments according to biological and molecular tumor's characteristics. CLINICAL TRIAL REGISTRATION: This study is registered under NCT02408536 on ClinicalTrials.gov .

The Functional Characteristics of Goat Cheese Microbiota from a One-Health Perspective.

Goat cheese is an important element of the Mediterranean diet, appreciated for its health-promoting features and unique taste. A pivotal role in the development of these characteristics is attributed to the microbiota and its continuous remodeling over space and time. Nevertheless, no thorough study of the cheese-associated microbiota using two metaomics approaches has previously been conducted. Here, we employed 16S rRNA gene sequencing and metaproteomics to explore the microbiota of a typical raw goat milk cheese at various ripening timepoints and depths of the cheese wheel. The 16S rRNA gene-sequencing and metaproteomics results described a stable microbiota ecology across the selected ripening timepoints, providing evidence for the microbiologically driven fermentation of goat milk products. The important features of the microbiota harbored on the surface and in the core of the cheese mass were highlighted in both compositional and functional terms. We observed the rind microbiota struggling to maintain the biosafety of the cheese through competition mechanisms and/or by preventing the colonization of the cheese by pathobionts of animal or environmental origin. The core microbiota was focused on other biochemical processes, supporting its role in the development of both the health benefits and the pleasant gustatory nuances of goat cheese.

Arthralgia in patients with ovarian cancer treated with bevacizumab and chemotherapy.

BACKGROUND: Chemotherapy with carboplatin, paclitaxel, and bevacizumab is the standard therapy for patients with advanced stage ovarian cancer wild-type BRCA after primary surgery. The most frequent side effects of bevacizumab in this setting are hypertension, thrombosis, hemorrhage, and proteinuria, while arthralgia has been poorly described. OBJECTIVE: To examine the incidence, duration, and reversibility of arthralgia. PATIENTS AND METHODS: A retrospective analysis was performed to describe the occurrence and outcome of arthralgia in 114 patients with advanced ovarian cancer, given first-line treatment with a combination of carboplatin, paclitaxel, and bevacizumab. Statistical analysis was performed to investigate a possible prognostic role of arthralgia, with progression-free survival as endpoint. RESULTS: 47 of 114 patients (41%) developed arthralgia during therapy. All patients had grade 1 or grade 2 arthralgia. Toxicity persisted after the end of bevacizumab in 17/47 patients (36%). Median progression-free survival for patients without arthralgia was 18 months (95% CI 14 to 24) compared with 29 months (95% CI 21 to not reached) for patients experiencing arthralgia (p=0.03). In order to avoid possible biases related to treatment duration, a multivariable Cox proportional hazards model including toxicity as a time dependent variable and age, stage, and residual disease after primary surgery was performed. In this model no variable showed a statistically significant association with progression-free survival. CONCLUSION: A high incidence of arthralgia (41%) was found and although rogression-free survival was worse for those patients who developed arthralgia, this was not maintained on multivariate analysis. Guidelines for treatment of this adverse event are needed.

Effects of ecotrofin™ on milk yield, milk quality and serum biochemistry in lactating goats.

A nutritional supplement (Ecotrofin™, by Vetoquinol Italia S.r.l) recommended in ruminants feeding to strengthen the physiological condition and improve digestive performance was tested in 20 pluriparae grazing goats divided in two groups (control and treated) to assess its possible effects on milk yield and quality and to assess eventual adverse effects. Animals from both groups also received 400 g/day of corn meal, and the treated group was supplemented with 20 g/head/day of the nutritional supplement. At the doses suggested by the manufacturer, despite a transient increase after 30 days of supplementation, Ecotrofin™ did not show significant effects on milk yield and, although some changes were found in the fatty acids profile, no significant improvement of MUFA and PUFA, as well as of omega-6:omega-3 ratio and CLA content were seen. Therefore, in our experimental conditions the supplementation of diet with Ecotrofin™ did not appear useful to improve goat's performance. A significant effect on kidney health markers (27 vs. 22.5 for urea and 0.83 vs. 0.76 for creatinine, p < 0.05) suggested a beneficial effect on renal function but, since levels fell in the normal ranges in both groups, such hypothesis would need further studies to be addressed.

Does Pelleted Starter Feed Restriction and Provision of Total Mixed Ration Ad Libitum during Weaning Influence the Behavior of Dairy Calves?

Currently, in dairy farming, there is growing concern for the welfare of calves during the critical period between the separation from their dams and weaning. During weaning, rationed feeding is a practice used to improve feed efficiency and control the calves' growth, but it could also have negative consequences associated with hunger and feed restriction behavior. One such consequence could be the performance by calves of stereotyped behaviors indicative of poor welfare, such as non-nutritive oral behaviors. We hypothesized that making a Total Mixed Ration (TMR) available to calves, in addition to the standard pelleted starter diet, thanks to its structural and nutritional characteristics, could help to focus the oral behavior of the subjects towards nutritional activities and therefore limit the development of stereotyped behaviors, even when the amount of starter is restricted. To test this hypothesis, 30 female Holstein calves (equipped with an accelerometer based on an ear tag), were randomly assigned to one of three treatments: a control diet (CTR) with an ad libitum calf starter but without TMR; Treatment 1 (TRT1) with both ad libitum calf starter and ad libitum TMR; Treatment 2 (TRT2) with ad libitum TMR and a restricted amount of a calf starter (50% of the intake starter of the control group day by day). All animals were kept in individual cages equipped with a slow-flow teat bucket apparatus for milk feeding and with access to separated buckets (one for drinking water, one for the starter, and one for the TMR) placed on the outside of the front gate of each cage. Sucking behavior, as well as resting, ruminating and activity behavior, was recorded individually from 7 days of life to weaning (63 d of age) by an automated monitoring system based on ear-tag accelerometers (SCR eSense, Allflex, Irving, TX, USA). The results showed that in the CTR group, there was a greater sucking activity compared to the TRT1 group (26.25 min/head/day vs. 16.83 min/head/day, p = 0.0181), while the TRT2 group showed intermediate values (20.41 min/head/day). We hypothesized that this increased sucking activity may be explained by the oral activity directed at sucking the Milk Bar tube observed only in CTR calves after complete consumption of available milk and could indicate a higher stress level than in the TRT1 group. The time spent resting was significantly lower for the CTR group compared to the TRT1 group (9.20 h/day vs. 11.15 h/day, p = 0.0049) while the TRT2 group was in an intermediate situation (10.65 h/day). Furthermore, the increase in time dedicated to rest observed in TRT1, in light of the good vitality of the calves shown by the daily patterns of resting behavior and daily activity, also seems to suggest an improvement in welfare conditions compared to calves receiving pelleted starter alone. Based on these results, we can deduce that providing TMR ad libitum in the diet of weaning calves could be an effective strategy to improve calf welfare due to the reduction in stereotyped oral activity. However, further studies are needed to establish threshold values useful for distinguishing physiological from pathological resting times in weaning calves raised in individual cages.

Darts fast-learning reduces theta power but is not affected by Hf-tRNS: A behavioral and electrophysiological investigation.

Sports trainers have recently shown increasing interest in innovative methods, including transcranial electric stimulation, to enhance motor performance and boost the acquisition of new skills during training. However, studies on the effectiveness of these tools on fast visuomotor learning and brain activity are still limited. In this randomized single-blind, sham-controlled, between-subjects study, we investigated whether a single training session, either coupled or not with 2 mA online high-frequency transcranial random noise stimulation (hf-tRNS) over the bilateral primary motor cortex (M1), would affect dart-throwing performance (i.e., radial error, arm range of motion, and movement variability) in 37 healthy volunteers. In addition, potential neurophysiological correlates were monitored before and after the training through a 32-electrode portable electroencephalogram (EEG). Results revealed that a single training session improved radial error and arm range of motion during the dart-throwing task, but not movement variability. Furthermore, after the training, resting state-EEG data showed a decrease in theta power. Radial error, arm movement, and EEG were not further modulated by hf-tRNS. This indicates that a single training session, regardless of hf-tRNS administration, improves dart-throwing precision and movement accuracy. However, it does not improve movement variability, which might require multiple training sessions (expertise resulting in slow learning). Theta power decrease could describe a more efficient use of cognitive resources (i.e., attention and visuomotor skills) due to the fast dart-throwing learning. Further research could explore different sports by applying longer stimulation protocols and evaluating other EEG variables to enhance our understanding of the lasting impacts of multi-session hf-tRNS on the sensorimotor cortex within the framework of slow learning and training assistance.

Detection of Antimicrobial Proteins/Peptides and Bacterial Proteins Involved in Antimicrobial Resistance in Raw Cow's Milk from Different Breeds.

Proteins involved in antibiotic resistance (resistome) and with antimicrobial activity are present in biological specimens. This study aims to explore the presence and abundance of antimicrobial peptides (AMPs) and resistome proteins in bovine milk from diverse breeds and from intensive (Pezzata rossa, Bruna alpina, and Frisona) and non-intensive farming (Podolica breeds). Liquid atmospheric pressure matrix-assisted laser desorption/ionization (LAP-MALDI) mass spectrometry (MS) profiling, bottom-up proteomics, and metaproteomics were used to comprehensively analyze milk samples from various bovine breeds in order to identify and characterize AMPs and to investigate resistome proteins. LAP-MALDI MS coupled with linear discriminant analysis (LDA) machine learning was employed as a rapid classification method for Podolica milk recognition against the milk of other bovine species. The results of the LAP-MALDI MS analysis of milk coupled with the linear discriminant analysis (LDA) demonstrate the potential of distinguishing between Podolica and control milk samples based on MS profiles. The classification accuracy achieved in the training set is 86% while it reaches 98.4% in the test set. Bottom-up proteomics revealed approximately 220 quantified bovine proteins (identified using the Bos taurus database), with cathelicidins and annexins exhibiting higher abundance levels in control cows (intensive farming breeds). On the other hand, the metaproteomics analysis highlighted the diversity within the milk's microbial ecosystem with interesting results that may reflect the diverse environmental variables. The bottom-up proteomics data analysis using the Comprehensive Antibiotic Resistance Database (CARD) revealed beta-lactamases and tetracycline resistance proteins in both control and Podolica milk samples, with no relevant breed-specific differences observed.

Spectral Profiling (Fourier Transform Infrared Spectroscopy) and Machine Learning for the Recognition of Milk from Different Bovine Breeds.

The Podolica cattle breed is widespread in southern Italy, and its productivity is characterized by low yields and an extraordinary quality of milk and meats. Most of the milk produced is transformed into "Caciocavallo Podolico" cheese, which is made with 100% Podolica milk. Fourier Transform Infrared Spectroscopy (FTIR) is the technique that, in this research work, was applied together with machine learning to discriminate 100% Podolica milk from contamination of other Calabrian cattle breeds. The analysis on the test set produced a misclassification percentage of 6.7%. Among the 15 non-Podolica samples in the test set, 2 were misclassified and recognized as Podolica milk even though the milk was from other species. The correct classification rate improved to 100% when the same method was applied to the recognition of Podolica and Pezzata Rossa milk produced by the same farm. Furthermore, this technique was tested for the recognition of Podolica milk mixed with milk from other bovine species. The multivariate model and the respective confusion matrices obtained showed that all the 14 Podolica samples (test set) mixed with 40% non-Podolica milk were correctly classified. In addition, Pezzata Rossa milk produced by the same farm was detected as a contaminant in Podolica milk from the same farm down to concentrations as little as 5% with a 100% correct classification rate in the test set. The method described yielded higher accuracy values when applied to the discrimination of milks from different breeds belonging to the same farm. One of the reasons for this phenomenon could be linked to the elimination of the environmental variable. However, the results obtained in this work demonstrate the possibility of using FTIR to discriminate between milks from different breeds.

Harmonization of homologous recombination deficiency testing in ovarian cancer: Results from the MITO16A/MaNGO-OV2 trial.

BACKGROUND: Homologous Recombination Deficiency (HRD) status predicts response to treatment with poly(ADP-ribose) polymerase inhibitors in Ovarian Cancer (OC) patients. The Myriad myChoiceCDx Assay is approved by Food and Drug Agency for the HRD assessment. Here we compared the HRD status obtained by three commercial panels with the results from Myriad reference test. METHODS: The HRD analysis was performed on DNA from formalin-fixed and paraffin-embedded tumor samples of 100 untreated OC patients for which Myriad assay results were available, using TruSight Oncology 500 HRD assay (Illumina), Oncomine Comprehensive Assay Plus (Thermo Fisher Scientific) and SOPHiA DDM HRD solution panel (SOPHiA Genetics). RESULTS: A good overall concordance with the reference method was demonstrated at three different levels: BRCA mutational status (from 94.4 % to 97.7 %), the genomic instability value (from 88.2 % to 95.3 %) and for the HRD status (from 90.4 % to 97.6 %). Moreover, a trend in favour of HRD positive patients for response rate, progression-free survival and overall survival similar to Myriad was observed for all three tests. DISCUSSION: Our data suggest the feasibility of commercial testing for assessing HRD status, with a good concordance with the reference method and association with clinical outcome.

Recent Advances in the Determination of Milk Adulterants and Contaminants by Mid-Infrared Spectroscopy.

The presence of chemical contaminants, toxins, or veterinary drugs in milk, as well as the adulteration of milk from different species, has driven the development of new tools to ensure safety and quality. Several analytical procedures have been proposed for the rapid screening of hazardous substances or the selective confirmation of the authenticity of milk. Mid-infrared spectroscopy and Fourier-transform infrared have been two of the most relevant technologies conventionally employed in the dairy industry. These fingerprint methodologies can be very powerful in determining the trait of raw material without knowing the identity of each constituent, and several aspects suggest their potential as a screening method to detect adulteration. This paper reviews the latest advances in applying mid-infrared spectroscopy for the detection and quantification of adulterants, milk dilution, the presence of pathogenic bacteria, veterinary drugs, and hazardous substances in milk.

The Effect of Feeding a Total Mixed Ration with an ad libitum or Restricted Pelleted Starter on Growth Performance, Rumination Behavior, Blood Metabolites, and Rumen Fermentation in Weaning Holstein Dairy Calves.

The aim of the current study was to evaluate the effect of the starter restriction and of the ad libitum TMR (total mixed ration) inclusion on intake, growth performance, rumination time (RT), and health condition of Holstein dairy calves during weaning. We randomly assigned thirty female Holstein calves (with an average weight of 38.5 ± 1.96 kg at birth) to one of three treatments. From 21 days of age, the calves were fed one of three treatments as follows: a control diet (CTR) with an ad libitum calf starter but without TMR; Treatment 1 diet (TRT1) with both an ad libitum calf starter and ad libitum TMR; Treatment 2 diet (TRT2) with ad libitum TMR and a restricted amount of a calf starter (50% of the intake recorder in the control group day by day). Calves in the TRT2 group, between 56 and 63 days of age, had a lower body weight (80.1; 79.5; 75.6 kg for the CTR, TRT1, and TRT2 groups, respectively) compared with CTR and TRT1 calves. This outcome is ascribed to the average daily gain (0.759; 0.913; 0.508 kg/day for the CTR, TRT1, and TRT2 groups, respectively), resulting also in TRT2 being lower than CTR or TRT1 calves. The inclusion of ad libitum TMR increased the rumination time, especially after weaning (15.28 min/h, 18.38 min/h, and 18.95 min/h for the CTR, TRT1, and TRT2 groups, respectively). Concerning the rumen metabolism and inflammometabolic response, overall, no differences were observed between the three dietary treatments. In conclusion, the results indicated that a TMR could partially replace a calf starter in weaning dairy calves, since neither growth performance nor health status were impaired. In addition, providing TMR (with or without concentrate restriction) led to a better rumen development and likely a better rumen fermentation efficiency in post-weaning.

Biological and clinical impact of membrane EGFR expression in a subgroup of OC patients from the phase IV ovarian cancer MITO-16A/MANGO-OV2A trial.

BACKGROUND: Validated prognostic biomarkers for anti-angiogenic therapy using the anti-VEGF antibody Bevacizumab in ovarian cancer (OC) patients are still an unmet clinical need. The EGFR can contribute to cancer-associated biological mechanisms in OC cells including angiogenesis, but its targeting gave disappointing results with less than 10% of OC patients treated with anti-EGFR compounds showing a positive response, likely due to a non adequate selection and stratification of EGFR-expressing OC patients. METHODS: EGFR membrane expression was evaluated by immunohistochemistry in a cohort of 310 OC patients from the MITO-16A/MANGO-OV2A trial, designed to identify prognostic biomarkers of survival in patients treated with first line standard chemotherapy plus bevacizumab. Statistical analyses assessed the association between EGFR and clinical prognostic factors and survival outcomes. A single sample Gene Set Enrichment-like and Ingenuity Pathway Analyses were applied to the gene expression profile of 195 OC samples from the same cohort. In an OC in vitro model, biological experiments were performed to assess specific EGFR activation. RESULTS: Based on EGFR-membrane expression, three OC subgroups of patients were identified being the subgroup with strong and homogeneous EGFR membrane localization, indicative of possible EGFR out/in signalling activation, an independent negative prognostic factor for overall survival of patients treated with an anti-angiogenic agent. This OC subgroup resulted statistically enriched of tumors of histotypes different than high grade serous lacking angiogenic molecular characteristics. At molecular level, among the EGFR-related molecular traits identified to be activated only in this patients' subgroup the crosstalk between EGFR with other RTKs also emerged. In vitro, we also showed a functional cross-talk between EGFR and AXL RTK; upon AXL silencing, the cells resulted more sensitive to EGFR targeting with erlotinib. CONCLUSIONS: Strong and homogeneous cell membrane localization of EGFR, associated with specific transcriptional traits, can be considered a prognostic biomarker in OC patients and could be useful for a better OC patients' stratification and the identification of alternative therapeutic target/s in a personalized therapeutic approach.

Mid-infrared (MIR) spectroscopy for the detection of cow's milk in buffalo milk.

In Italy, buffalo mozzarella is a largely sold and consumed dairy product. The fraudulent adulteration of buffalo milk with cheaper and more available milk of other species is very frequent. In the present study, Fourier transform infrared spectroscopy (FTIR), in combination with multivariate analysis by partial least square (PLS) regression, was applied to quantitatively detect the adulteration of buffalo milk with cow milk by using a fully automatic equipment dedicated to the routine analysis of the milk composition. To enhance the heterogeneity, cow and buffalo bulk milk was collected for a period of over three years from different dairy farms. A total of 119 samples were used for the analysis to generate 17 different concentrations of buffalo-cow milk mixtures. This procedure was used to enhance variability and to properly randomize the trials. The obtained calibration model showed an R 2 ≥ 0.99 (R 2cal. = 0.99861; root mean square error of cross-validation [RMSEC] = 2.04; R 2val. = 0.99803; root mean square error of prediction [RMSEP] = 2.84; root mean square error of cross-validation [RMSECV] = 2.44) suggesting that this method could be successfully applied in the routine analysis of buffalo milk composition, providing rapid screening for possible adulteration with cow's milk at no additional cost.

The Quality of Five Natural, Historical Italian Cheeses Produced in Different Months: Gross Composition, Fat-Soluble Vitamins, Fatty Acids, Total Phenols, Antioxidant Capacity, and Health Index.

Five natural historic cheeses of Southern Italy were investigated-Caciocavallo Palermitano (CP), Casizolu del Montiferru (CdM), Vastedda della Valle del Belìce (VVB), Pecorino Siciliano (PS), and Caprino Nicastrese (CN)-which are produced with raw milk and with traditional techniques and tools, from autochthonous breeds reared under an extensive system. The effects of the month of production on gross composition, MUFA, PUFA, PUFA-ω6, PUFA-ω3, α-tocopherol, retinol, cholesterol, TPC, TEAC, and GHIC were evaluated. In CP, CLA, TPC, and GHIC were higher in April than in February. CdM showed higher values in terms of fat, saturated fatty acids, PUFA-ω3, α-tocopherol, TEAC, and GHIC in May than in February and September, while low values in terms of protein, moisture, and CLA were found. In VVB, MUFA, PUFA-ω6, and α-tocopherol increased in June compared with April; conversely, protein, FRAP, and TEAC were higher in April. In PS, protein, CLA, PUFA, PUFA-ω3, α-tocopherol, and GHIC increased in May compared with January; on the contrary, moisture, NaCl, and TEAC showed high values in January. CN showed higher values in terms of PUFA, PUFA-ω6, PUFA-ω3, TPC, TEAC, and GHIC in April and June compared with January. It is shown that each cheese is unique and closely linked to the production area. Cheeses produced in the spring months showed a high nutritional quality due to the greatest presence of healthy compounds originating from an extensive feeding system.

Analysis of A Disintegrin and Metalloprotease 17 (ADAM17) Expression as a Prognostic Marker in Ovarian Cancer Patients Undergoing First-Line Treatment Plus Bevacizumab.

To find prognostic factors for advanced ovarian cancer patients undergoing first-line therapy with carboplatin, paclitaxel and bevacizumab, we investigated the expression of a disintegrin and metalloprotease 17 (ADAM17) in cancer tissues. ADAM17 has been involved in ovarian cancer development, progression and cell resistance to cisplatin. Tissue microarrays from 309 ovarian cancer patients enrolled in the MITO16A/MANGO-OV2 clinical trial were analyzed by immunohistochemistry for ADAM17 protein expression. Intensity and extent of staining were combined into a semi-quantitative visual grading system (H score) which was related to clinicopathological characteristics of cases and the clinical outcome of patients by univariate and multivariate Cox regression models. ADAM17 immunostaining was detected in most samples, mainly localized in the tumor cells, with variable intensity across the cohort. Kaplan-Meier survival curves, generated according to the best cut-off value for the ADAM17 H score, showed that high ADAM17 expression was associated with worse prognosis for PFS and OS. However, after the application of a shrinkage procedure to adjust for overfitting hazard ratio estimates, the ADAM17 value as prognostic factor was lost. As subgroup analysis suggested that ADAM17 expression could be prognostically relevant in cases with no residual disease at baseline, further studies in this patient category may be worth planning.

Biological Role of Tumor/Stromal CXCR4-CXCL12-CXCR7 in MITO16A/MaNGO-OV2 Advanced Ovarian Cancer Patients.

This study investigated the prognostic role of the CXCR4-CXCL12-CXCR7 axis in advanced epithelial ovarian cancer (EOC) patients receiving first-line treatment within the MITO16A/MaNGO-OV2 phase-IV trial. CXCR4-CXCL12-CXCR7 expression was evaluated in the epithelial and stromal component of 308 EOC IHC-stained tumor samples. The statistical analysis focused on biomarkers' expression, their association with other variables and prognostic value. Zero-inflated tests, shrinkage, bootstrap procedures, and multivariable models were applied. The majority of EOC (75.0%) expressed CXCR4 and CXCR7, 56.5% expressed the entire CXCR4-CXCL12-CXCR7 axis, while only 4.6% were negative for CXCL12 and its cognate receptors, in regard to the epithelial component. Stromal CXCL12 and CXCR7, expressed in 11.2% and 65.5%, respectively, were associated with the FIGO stage. High CXCL12 in epithelial cancer cells was associated with shorter progression-free and overall survival. However, after adjusting for overfitting due to best cut-off multiplicity testing, the significance was lost. This is a wide-ranging, prospective study in which CXCR4-CXCL12-CXCR7 were systematically evaluated in epithelial and stromal components, in selected stage III-IV EOC. Although CXCL12 was not prognostic, epithelial expression identified high-risk FIGO stage III patients for PFS. These data suggest that it might be worth studying the CXCL12 axis as a therapeutic target to improve treatment efficacy in EOC patients.

Suitability of Protein Content Measured by MilkoScan FT-Plus Milk Analyzer to Evaluate Bovine and Ovine Colostrum Quality.

The objective of this study was to evaluate MilkoScan FT-plus for the estimation of the immunoglobulin G (IgG) content in bovine and ovine colostrum. Between April and May 2016, a total of 94 colostrum samples (54 from Simmental dairy cows and 39 from Sarda ewes) were collected within 6 h (T0) and after 24 h (T24) from parturition. Colostrum samples were subjected to the radial immunodiffusion (RID) assay for the quantification of IgG and to MilkoScan FT-plus for the estimation of protein content (TP, %), which was then used as an indirect method for the evaluation of colostrum quality. To compare the two methods, correlation and regression analysis of IgG quantification by RID and protein (%) content estimation by MilkoScan FT-plus data was performed using Procedure CORR and Procedure REG of SAS, respectively (version 9.3, SAS Institute Inc., Cary, NC, USA). Thresholds for the classification of good colostrum quality (as determined by RID assay, the gold standard method) were set at 50 g of IgG/L in cows and 20 g of IgG/L in ewes. The concentration of IgG in bovine colostrum assayed by RID showed a variation ranging from 41.45 to 199.97 g/L with an average of 99.85 ± 40.84 g/L at T0, and from 2.83 to 75.93 g/L with an average of 19.76 ± 19.01 g/L at T24. Regarding ovine colostrum, the concentration of IgG assayed by RID ranged from 34.45 to 156.32 g/L with an average value of 77.82 ± 37.58 g/L at T0, and from 5.6 to 69.74 g/L with an average of 27.90 ± 19.81 g/L at T24. Colostrum TP ranged from 3.70 to 23.96% for bovine colostrum and 6.32 to 22.88% for ovine colostrum using MilkoScan FT-plus. MilkoScan FT-plus and RID data were highly and significantly correlated (r = 0.91 for bovine and r = 0.94 for ovine colostrum), and regression analysis showed a strong relationship between IgG concentration provided by RID assay and TP provided by MilkoScan FT-plus (R2 = 0.84 and 0.88 for bovine and ovine, respectively). Optimal cut-off points for the greatest accuracy of TP (%) determined by MilkoScan FT-plus were 12.8% in cows [with 88.9% sensitivity (Se) and 100% specificity (Sp)] and 9% in ewes (with 96.7% Se and 100% Sp). In conclusion, these outcomes indicate that MilkoScan FT-plus as an indirect method may be a reliable tool for the estimation of the total IgG concentration and quality in bovine and ovine colostrum. Moreover, the cut-off levels of 12.8% for bovine and 9% for ovine of TP, seem sufficient to ensure that all poor-quality colostrum can be classified as such, with only a low proportion of good-quality colostrum being misclassified as poor-colostrum, thereby increasing the probability of delivering good-quality colostrum to new-born calves and lambs.