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The organic residues from agro-food sector can be transformed into valuable economic resources rather than representing an unmanageable waste, whose disposal in landfills contributes to increase both soil pollution and the greenhouse effect. With this in mind, the aim of the investigation is to design eco-friendly compatible innovative biological processes for producing lactic acid (LA) from fermentable sugars using as substrate tomato pomace. In the first part of the investigation, we show the physico-chemical characterization of tomato pomace produced in the 2017 season with or without thermal pre-treatment. pH, total and volatile solids content, sugars and cellulose are measured. The microorganism consortia suitable for lactic acid production are selected from buffalo dung, and compared with commercial strains of lactic acid producing bacteria. Both bacteria consortia are used as inoculum for the anaerobic fermentation of tomato pomace as sole substrate for producing fermentable sugars and convert them in LA. Sugars are measured by a spectrophotometric- coupled enzyme assay and LA by HPLC with electrical conductivity detector. The molecular analysis of the isolated bacteria is performed by using the denaturing gradient gel electrophoresis (DGGE). Anaerobic fermentation of tomato pomace is also performed using different initial pH values of the substrate. The final goal is to design biological processes aimed at achieving the highest LA recovery yields.

Industry has the severe problem to accumulate, treat and dispose food processing wastes and by-products. The main objective of many recent studies is to test innovative processes and technologies for recycling and upgrading these wastes for recovering high-value-added marketable materials in compliance with the European Sustainable Waste Management Guidelines and the principles of the "circular economy”. In particular, new, innovative and profitable management strategies for sludgy waste from tomato processing industries could be fostered in Campania. Alternative and valuable process chains are possible in which different players working together to “eco-friendly” recycle, reuse and promote tomato industry residual biomasses could obtain high value-added products with a sustainable approach. One of the possible added value product that can be obtained by tomato industrial processing waste is lactic acid, whose production could have a remarkable economic and ecological impact. In this study we provide an estimate of the available tomato industry processing waste and related potential for lactic acid production in Campania, carrying out a survey on the more suitable production technologies. Maximum radius distance of industries from a possible/hypothetical centralized transformation plant according to the biomass availability and transportation facilities is calculated. Feasibility studies of eco-friendly processes able to convert tomato processing waste in lactic acid are considered, evaluating the actual amount, reuse or disposal in waste dumps or soil of the tomato pomace. The benefits of the correct management of tomato processing waste is considered, evaluating the possible economic revenue resulting from the proposed system.

In this paper we study the anaerobic digestion of water buffalo manure collected during a period of three years with the goal of investigating the effect of the substrate C/N ratio on the bio-methane production. The investigated samples show a C/N ratio between 9 and 50, this interval being much wider than that typically considered optimal in the literature for the waste digestion processes. The experimental tests are performed in batch mode, at 37 °C and with a starting pH equal either to 6 or 7, uncontrolled during the digestion process. We observe that, regardless of the C/N ratio of tested samples, the Gompertz productivity, expressed in terms of bio-methane volume fraction, is always larger than 54 %. A slightly larger productivity is recorded for substrates digested starting from an initial pH set to 7. We may then conclude that the C/N ratio of water buffalo manure does not really affect the bio-methane yield.

Animal manure can be profitably used to produce energy in anaerobic fermenters increasing energy independence and reducing disposal costs and pollution. Animal manure is a low cost substrate rich in carbohydrates, especially suitable for biofuels production, such as hydrogen and methane. We preliminary studied the effect of mechanical manure pre-treatments on the fermentation process focusing on the biogas yield and composition. In particular, we investigated both the H2 and CH4 oriented fermentation. Our batch reactors consist in 280 ml serum glass bottles hermetically sealed with pierceable rubber cups. The products of fermentation were measured with a micro gas chromatograph, equipped with two capillary columns. To individuate the most suitable conditions for H2 production, a manure thermal pre-treatment, used to isolate the H2-producing bacteria, was investigated. Preliminary results show that the mechanical pre-treatments do not systematically alter the fermentation process. Concerning the thermal pre-treatment, we found that the CH4 production is inhibited when the manure is heated at the 90 °C for several hours. Moreover, we identified the most favorable conditions to optimize the production of H2: i) thermal pre-treatment on the substrate; ii) initial pH ~ 5.0; iii) process temperature of 55 °C. The favorable conditions to optimize the production of CH4 are: i) initial pH ~ 7.0;ii) process temperature of 37 °C.

Animal manure is a low cost substrate suitable for biofuels production, such as hydrogen and methane, via anaerobic fermentation (Kyazze et al., 2007). Previous studies attributed the higher biogas yield from certain types of manure to the presence of a native microflora (Yeole and Ranande, 1992) and/or a higher or better carbon-nitrogen (C/N) ratio (Fulford, 1988). Probably the main difference between the different types of dung is related to the specific food the animal eat, which changes the overall C content and C/N ratio. We measured the C/N ratio and the native microflora of dung collected during one year. We related it to the biogas yield during anaerobic fermentation both for lactating and non-lactating buffaloes manure. The results showed that the buffalo manure was in both cases plenty of carbohydrates with a C percentage content based on the dry sample weight of 36.2 ± 2.0 % and 31.4 ± 1.5 %, respectively. The C/N ratio was 18.6 ± 1.5 % and 20.6 ± 2.9 %, respectively, and it changed during the different seasons because of the fluctuating N content. Indeed we found an acceptable correlation (R = 0.75) between the average environmental temperature and the C/N ratio of lactating buffalo manure. Neither the C or N content nor the C/N ratio appeared to be strongly correlated to the biogas production. The manure was fermented in batch reactors at 37 °C and pH = 6.0 after different pretreatments of the collected manure. The analysis of the microbial community was carried out using denaturing gradient gel electrophoresis (DGGE) to generate fingerprints of 16S rRNA genes (Carillo et al., 2012). The sequences analysis revealed that the Eubacterial community during fermentation in batch reactors was much more taxon rich and diversified than the Archeal community. The study of Archeal community instead showed the presence of only three main methanogen strains very active in methane production and largely predominant.

In this work hydrogen and methane production from buffalo manure has been scrutinized, by means of the anaerobic dark fermentation process and different process conditions. Animal manure characteristics are largely variable during different periods of the year because of the unavoidable changes in animal food supply and digestion apparatus of buffalo. A very significant variability was observed for water buffaloes manure, collected from the same supplier. This implies also a large variability in biogas production by anaerobic fermentation of such substrates. To overcome this variability, we mixed fresh with digested manure. This procedure is consolidated as regard methane production. We also investigated a similar strategy to optimize and stabilize H2 yield. We were able to find optimal process conditions for hydrogen and methane production with this type of substrate. The experiments have been run in batch mode. The products of fermentation, H2, CO2 and CH4, have been measured with a micro-gas chromatograph. Moreover the potential production of H2 has been maximized by thermally pre-treating the substrate and using a process temperature of the fermenter equal to 55 °C and an initial pH value of 5.5, while very high levels of potential production of CH4 have been obtained at 37 °C and with an initial pH of 7.0. The effect of different strategies has also been analyzed to highlight the different microbial community present in the substrate using the denaturing gradient gel electrophoresis (DGGE). This investigation has been carried out with the aim at isolating and selecting of natural bacterial species to use as inoculum to improve and stabilize the fermentation process of buffalo manure. More results concerning the microbial communities present in the substrate and responsible for biogas production are presented in a companion paper. Here we only list the main types of bacteria identified in the manure.

Abstract In this work, an intercomparison of sensitization effects produced by gold (GNP) and dextran-coated iron oxide (SPION-DX) nanoparticles in M059J and U87 human glioblastoma cells was performed using 6 MV-photons. Three variables were mapped: the nanoparticle material, treatment concentration, and cell radiosensitivity. For U87, GNP treatments resulted in high sensitization enhancement ratios (SER $$_{10\%}$$ 10 % up to 2.04). More modest effects were induced by SPION-DX, but still significant reductions in survival were achieved (maximum SER $$_{10\%}=1.61$$ 10 % = 1.61 ). For the radiosensitive M059J, sensitization by both NPs was poor. SER $$_{10\%}$$ 10 % increased with the degree of elemental uptake in the cells, but not necessarily with treatment concentration. For GNP, where exposure concentration and elemental uptake were found to be proportional, SER $$_{10\%}$$ 10 % increased linearly with concentration in both cell lines. For SPION-DX, saturation of sensitization enhancement and metal uptake occurred at high exposures. Fold change in the $$\alpha /\beta$$ α / β ratios extracted from survival curves are reduced by the presence of SPION-DX but strongly increased by GNPs , suggesting that sensitization by GNPs occurs mainly via promotion of lethal damage, while for SPION-DX repairable damage dominates. The NPs were more effective in eliminating the radioresistant glioblastoma cells, an interesting finding, as resistant cells are key targets to improve treatment outcome.

The release of damage-associated molecular patterns, including uridine triphosphate (UTP) and adenosine triphosphate (ATP) to the extracellular milieu is a key component of innate immune response to infection. Previously, we showed that macrophage infection by the protozoan parasite Leishmania amazonensis—the etiological agent of cutaneous leishmaniasis—can be controlled by ATP- and UTP-mediated activation of P2Y and P2X7 receptors (activated by UTP/ATP and ATP, respectively), which provided comparable immune responses against the parasite. Interestingly, in context of Leishmania amazonensis infection, UTP/P2Y triggered apoptosis, reactive oxygen species, and oxide nitric (NO) production, which are characteristic of P2X7 receptor activation. Here, we examined a possible “cross-talk” between P2Y2 and P2X7 receptors, and the requirement for pannexin-1 (PANX-1) in the control of L. amazonensis infection in mouse peritoneal macrophages and in vivo. UTP treatment reduced L. amazonensis parasite load, induced extracellular ATP release [which was pannexin-1 (PANX-1) dependent], and triggered leukotriene B4 (LTB4) production in macrophages. UTP-induced parasite control was blocked by pharmacological antagonism of P2Y2 or P2X7 receptors and was absent in macrophages lacking P2X7 or PANX-1. In addition, ATP release induced by UTP was also inhibited by PANX-1 blocker carbenoxolone, and partially reversed by inhibitors of vesicle traffic and actin cytoskeleton dynamics. In vivo, UTP treatment reduced footpad and popliteal lymph node parasite load, and the lesion in wild-type (WT) mice; fact not observed in P2X7−/− mice. Our data reveal that P2Y2 and P2X7 receptors cooperate to trigger potent innate immune responses against L. amazonensis infection.

Abstract Glioblastomas are the most common form of primary tumors of the central nervous system (CNS) and despite treatment, patients with these tumors have a very poor prognosis. ATP and other nucleotides and nucleosides are very important signaling molecule in physiological and pathological conditions in the CNS. ATP is degraded very slowly by gliomas when compared to astrocytes, potentially resulting in the accumulation of extracellular ATP around gliomas. Cell lysis caused by excitotoxic death or by tumor resection may liberate intracellular ATP, a known mitotic factor for glioma cells. The aim of this study is to examine the effects on cytotoxicity induced by extracellular ATP in U138-MG human glioma cell line and C6 rat glioma cell line compared to hippocampal organotypic cell cultures. The cytotoxicity of ATP (0.1, 0.5, 5 mM) was measured using propidium iodide and LDH assays. Caspases assay was performed to identify apoptotic cell death. Results showed that the glioma cells present resistance to death induced by ATP when compared with a normal tissue. High ATP concentrations (5 mM) induced cell death after 24 h in organotypic cell cultures but not in glioma cell lines. Our data indicate that ATP released in these situations can induce cell death of the normal tissue surrounding the tumor, potentially opening space to the fast growth and invasion of the tumor. [ABSTRACT FROM AUTHOR]

ABSTRACT Purpose: Testicular germ cells tumor (TGCT) are associated with a high cure rate and are treated with platinum-based chemotherapy. However, a group of testicular cancer patients may have a very unfavorable evolution and insensitivity to the main therapeutic agent chemotherapy (CT) cisplatin. The aim of this study was to evaluate the risk of recurrence and overall survival related to the expression of nuclear factor kappa-B (NF-κB), transglutaminase 2 (TG2) and excision repair cross-complementation group 1 (ERCC1) in patients with TGCT treated with platinum combinations. Patients and Methods: A retrospective study was performed with TGCT patients treated with platinum-based chemotherapy. Immunohistochemical analysis was performed and the expression was correlated with clinical and laboratory data. Results: Fifty patients were included, the mean age was 28.4 years (18 to 45), and 76% were non-seminoma. All patients were treated with standard cisplatin, etoposide and bleomycin or cisplatin, and etoposide. Patient’s analyzed immunodetection for NF-κB, TG2, and ERCC1 were positive in 76%, 54% and 42%, respectively. Multivariate analysis identified that positive expressions to ERCC1 and NF-κB are independent risk factors for higher recurrence TGCT after chemotherapy (RR 2.96 and 3.16, respectively). Patients with positive expression of ERCC1 presented a poor overall survival rate for 10-year follow (p=0.001). Conclusions: The expression of ERCC1 and NF-κB give a worse prognosis for relapse, and only ERCC1 had an influence on the overall survival of TGCT patients treated with platinum-based chemotherapy. These may represent markers that predict poor clinical outcome and response to cisplatin.

This study was aimed to characterize the depression-like behaviour in the classical model of chronic inflammation induced by Complete Freund's Adjuvant (CFA). Male Swiss mice received an intraplantar (i.pl.) injection of CFA (50 µl/paw) or vehicle. Behavioural and inflammatory responses were measured at different time-points (1 to 4 weeks), and different pharmacological tools were tested. The brain levels of IL-1β and BDNF, or COX-2 expression were also determined. CFA elicited a time-dependent edema formation and mechanical allodynia, which was accompanied by a significant increase in the immobility time in the tail suspension (TST) or forced-swimming (FST) depression tests. Repeated administration of the antidepressants imipramine (10 mg/kg), fluoxetine (20 mg/kg) and bupropion (30 mg/kg) significantly reversed depression-like behaviour induced by CFA. Predictably, the anti-inflammatory drugs dexamethasone (0.5 mg/kg), indomethacin (10 mg/kg) and celecoxib (30 mg/kg) markedly reduced CFA-induced edema. The oral treatment with the analgesic drugs dipyrone (30 and 300 mg/kg) or pregabalin (30 mg/kg) significantly reversed the mechanical allodyinia induced by CFA. Otherwise, either dipyrone or pregabalin (both 30 mg/kg) did not significantly affect the paw edema or the depressive-like behaviour induced by CFA, whereas the oral treatment with dipyrone (300 mg/kg) was able to reduce the immobility time in TST. Noteworthy, CFA-induced edema was reduced by bupropion (30 mg/kg), and depression behaviour was prevented by celecoxib (30 mg/kg). The co-treatment with bupropion and celecoxib (3 mg/kg each) significantly inhibited both inflammation and depression elicited by CFA. The same combined treatment reduced the brain levels of IL-1β, as well as COX-2 immunopositivity, whilst it failed to affect the reduction of BDNF levels. We provide novel evidence on the relationship between chronic inflammation and depression, suggesting that combination of antidepressant and anti-inflammatory agents bupropion and celecoxib might represent an attractive therapeutic strategy for depression.

Objective: To determine factors indicating testing frequency and positive test results in a Division I sports department intrapandemic., Design: Retrospective analysis., Setting: A single Division I collegiate sports department., Patients: All student-athlete (n = 437), student staff (n = 89), and adult staff (n = 202) members of the sports department. Total cohort (n = 728)., Interventions: The authors analyzed the independent variables of local positive rates, sport characteristics, and campus events for impact on the volume of the departmental testing and positive rates., Main Outcome Measures: Measured dependent variables of the volume of departmental testing and positive rates were analyzed., Results: Positive predictive rates (PPRs) largely differed from local, off-campus rates in timing and duration (59.52%: P < 0.05). Overall, 20633 tests were administered with 201 positive results (0.97% PPR). Student-athlete numbers were highest in all categories, followed by adult then student staff. Greater proportions of contact sports became positive (53.03%: P < 0.001) and all-male sports (47.69%: P < 0.001). No comparative difference was seen for teams using fomites (19.15%: P = 0.403). Spring sports teams had the lowest percentage of the team positive (22.22%: P < 0.001). Winter sports had the highest PPR (1.15%), all occurring during team-controlled activities. Playing sports indoors did not increase inside team-controlled activity positive rates ( P = 0.066)., Conclusions: Longitudinal changes in local, off-campus infection rates partially affected sports departmental positive results while testing rates were more influenced by sport and university schedule. Testing resources should be directed toward high-risk sports, which included contact sports (football, basketball, and soccer), all-male teams, both Winter and indoor sports inside team-controlled activities, and sports with long periods of time outside team-controlled activities., Competing Interests: The authors report no conflicts of interest., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

Some effects of expressing stereotypic behavior have not yet been elucidated. During gestation, the environment has the potential to interfere with offspring development and to have prenatal or longer-term consequences. We tested the hypothesis that the occurrence of stereotypic behavior during gestation could affect the phenotype of the offspring. Twenty-eight pregnant sows were studied by comparing two groups differing in the amount of stereotypy shown. We analyzed emotionality in the offspring from sows showing high or low stereotypy frequency using the open field and novel object tests. In the open field test, piglets from sows with a high rate of stereotypies walked more in central sectors ( p < 0.0001) and lateral sectors ( p = 0.04) than piglets from sows with a low rate of stereotypies. In the novel object test, the offspring from low stereotypy sows vocalized more ( p = 0.008). We demonstrate for the first time that the stereotypic behavior by the mother during gestation changes the phenotype of the offspring, in particular, their emotionality., (Copyright © 2020 Tatemoto, Bernardino, Morrone, Queiroz and Zanella.)

Manure from lactating and non-lactating water buffaloes was separately collected from a single dairy farm and anaerobically digested under mesophilic conditions in batch mode to produce biogas. This substrate, scarcely studied in the literature, showed two peculiarities regarding two fundamental parameters in the digestion processes: C/N ratio and initial pH. Typically, optimal C/N varies from 20 to 30, but in this work an almost negligible role of this ratio is observed. We demonstrated it by investigating a very large C/N interval, from 9.7 to 50.1, not by adding selected nutrients to the system, but exploiting the natural variation of the substrate. Concerning the pH, we show that also typically considered unfavorable conditions are feasible for this substrate. In fact, though neutral-basic initial pH is proved to be optimal to run the digestion process, in line with many other kinds of dungs, also acid initial pH leads to satisfactory CH 4 yield. This is principally related to the capability of water buffalo manure of auto-modifying the pH to neutrality during the digestion, when initial pH of 5.0 and 6.0 are considered. This aspect may be relevant in co-digestion processes with acid wastes, since it may allow not adding neither a buffer, nor a pH regulator to the system. All the digestion conditions are separately tested with lactating and non-lactating water buffaloes and no statistical meaningful differences exist between the two kinds of cattle., Competing Interests: Declaration of Competing Interest The authors declared that there is no conflict of interest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Aggressive interactions, and their consequences, are the most important causes of poor welfare in piglets. Aggressive behaviour can be modulated by the prenatal and neonatal environment in several species. Commercially kept pregnant sows are often subjected to food restriction, which can compromise their welfare. Limited information is available on the consequences of sow hunger during pregnancy on welfare outcomes for their piglets. High fibre diets can mitigate the feeling of hunger and, consequently, it may improve welfare and productivity measures. The aim of this study was to assess the consequences of feeding pregnant gilts with high fibre diets (HFD) on agonistic behaviour, as manifested by skin lesions, and indicators of fear in their piglets at weaning. Twenty-two pregnant gilts were fed either HFD, 12.86% of crude fibre, 2.4 kg per day (N = 14), or low fibre diet (LFD), 2.53% of crude fibre, 2.0 kg per day (N = 8). During lactation, both treatments received the same diet, ad libitum. We investigated the impact of HFD on behaviour and performance measures (birth weight, average daily gain, weaning weight, see S3 File) in the offspring. Skin lesions were evaluated before and after weaning in 156 piglets (100 HFD and 56 LFD), and 142 piglets were subjected to an open field test and a novel object test (87 HFD and 55 LFD). We found no treatment effect on the performance measures. Piglets born from gilts that received HFD had fewer skin lesions before weaning (D28) than the offspring of LFD gilts, while no difference was found during days 29 and 30. In the open field and novel object tests, there was no treatment effect on the behaviour of piglets. The improved skin health at weaning in piglets of sows fed HFD suggests less agonistic interactions amongst these littermates than in piglets of sows fed LFD., Competing Interests: The authors have declared that no competing interests exist.

Cytokines are cell signalling proteins that mediate a number of different physiological responses. They are also biomarkers for inflammatory conditions and potential diagnostic references for diseases. Until recently, simultaneous quantification of cytokine profiles had not been possible. Now however, fluorescent microsphere immunoassays (FMIA) are able to measure multiple cytokines in a single sample. The following pro-inflammatory and anti-inflammatory cytokines were quantified in equine plasma and serum samples: interleukin (IL)-2, IL-4, IL-6, IL-10, interferon (IFN)-γ, tumor necrosis factor (TNF)-α. •The objective of this study was to quantify six equine cytokines simultaneously using the BioPlex(®) 200 system in equine EDTA-plasma and serum.•It demonstrates an increased number of detectable cytokines over published studies.•This technology has the advantage of reduced sample volume and assay time compared to traditional sandwich ELISAs.

Buffalo dung is a low-cost substrate with plenty of carbohydrates, an optimal carbon/nitrogen ratio, and a rich microbial flora, and could become a valuable source of biogas. Therefore, in the present study we compared the type and amount of specific eubacteria to the different configurations of pH, temperature and thermal pretreatment after fermentation in batch reactors in order to understand the suitability of buffalo manure for hydrogen production. The phylogenetic structure of the microbial community in fermentation samples was studied using denaturing gradient gel electrophoresis to generate fingerprints of 16S rRNA genes. The sequences analysis revealed abundance of the phyla Bacteroidetes and Firmicutes, and in particular of the order Clostridiales. Very active hydrogen producing bacteria belonging to Clostridium cellulosi species were identified demonstrating the suitability of this substrate to produce hydrogen. Moreover, a large fraction of 16S-rDNA amplicons could not be assigned to lower taxonomic ranks, demonstrating that numerous microorganisms involved in anaerobic fermentation in digesters or bioreactors are still unclassified or unknown.