Animal products, in particular dairy and fermented products, are natural, major sources of lactic acid bacteria (LAB). Due to their antimicrobial properties, LAB are used in humans and in animals, with beneficial effects, as probiotics or in the treatment of a variety of diseases. In livestock production, LAB contribute to animal performance, health, and productivity. In the food industry, LAB are applied as bioprotective and biopreservation agents, contributing to improve food safety and quality. However, some studies have described resistance to relevant antibiotics in LAB, with the concomitant risks associated to the transfer of antibiotic resistance genes to foodborne pathogens, their potential dissemination throughout the food chain, and the environment. Here, we summarize the application of LAB in livestock and animal products, as well as the health impact of LAB in animal food products. In general, the beneficial effects of LAB on the human food chain seem to outweigh the potential risks associated with their consumption as part of animal and human diets. However, further studies and continuous monitorization efforts are needed to ensure their safe application in animal products and in the control of pathogenic microorganisms, preventing the possible risks associated with antibiotic resistance and, thus, protecting public health.

Posterior capsule opacification (PCO) is the most common complication of cataract surgery. It causes a gradual deterioration of visual acuity, which would otherwise remain improved after a successful procedure. Despite recent advances in ophthalmology, this complication has not been eradicated and the incidence of PCO can be as high as 10%. This article reviews the literature concerning the pathomechanism of PCO and examines the biochemical pathways involved in its formation and methods to prevent this complication. We also review the reported tests performed in cell cultures under laboratory conditions, in experimental animal models, and in ex vivo human lens capsules. Finally, we describe research involving human eyes in the clinical setting and pharmacological methods that may reduce the frequency of PCO. Due to the multifactorial eti-ology of PCO, in vitro studies make it possible to assess the factors contributing to its complica-tions and search for new therapeutic targets. Not all pathways involved in cell proliferation, mi-gration, and contraction of the lens capsule are reproducible in laboratory conditions; moreover, PCO in humans and laboratory animals may be additionally stimulated by various degrees of postoperative reactions depending on the course of surgery. Therefore, further studies are necessary.

3D cell cultures have improved the evaluation of drugs for cancer therapy, due to their high similarity to solid tumors. In melanoma, autophagy appears to show a dual role depending on the progression of the disease. p62 protein has been proposed for the evaluation of autophagic flux since its expression is an indicator of the state of autophagy. Pentoxifylline (PTX) and Norcantharidin (NCTD) are drugs that have been shown to have anticancer effects in various types of cancer. In this work, B16F1 mouse melanoma cell line was used, from which two-dimensional (2D) monolayer cultures and three-dimensional (3D) organoids were carried out using the hanging drop technique. The expression of p62 was determined by western blot and immunofluorescences. Our results indicate that pentoxifylline reduces p62 expression in both types of culture. While Norcantharidin increases its expression in 3D cultures at 24 h. Therefore, these drugs could have a potential therapeutic use for the regulation of autophagy in melanoma, depending on the state of evolution of the disease.

Bovine whey cheese (WC) is a product from southern European countries that presents some challenges: their elaboration process involves high energy inputs, the yield is low and WC has a short shelf-life. The application of ultrafiltration to bovine whey before its manufacture and the employment of protective cultures can reduce these disadvantages. The aim of this work was to manufacture whey cheeses using ultrafiltrated (UF) concentrated bovine cheese whey with added probiotics or probiotics plus protective cultures. Three types of WC were produced: control, without any addition (C); with the addition of the probiotic Lactobacillus acidophilus (LA5); with the addition of Lactobacillus acidophilus plus a protective culture containing Lactobacillus paracasei and Lactobacillus rhamnosus (LA5FQ4). The WCs were stored under refrigerated conditions for 28 days. The products with added cultures presented a lower pH and a higher titratable acidity when compared to the control. Sample LA5 presented the lowest pH and the highest titratable acidity while LA5FQ4 presented intermediate values. Slight differences were observed between products regarding color parameters, chiefly as a result of storage time. The samples with added cultures were firmer when compared to the control, showing LA5 cheeses the highest values at the end of the storage. Lactobacilli and lactococci counts were of the order of log 8-9 CFU/g for the products with added cultures. Lower levels of yeasts and moulds were detected on the sample with the protective culture (LA5FQ4), that by the end of storage presented counts one log cycle lower than C and LA5. Hence, it is evident the beneficial impact of the protective culture on the shelf life of the product. In what concerns to sensory evaluation, LA5FQ4 cheeses obtained the highest scores regarding all parameters evaluated. It could be concluded that the use of UF associated to the addition of protective cultures can be very useful to reduce the energy consumption of the process, prolong the shelf life of WC, and to improve its sensory properties.

Many dairy industries use their own starter cultures rather than Direct vat set culture because of their novelty and specific sensory characteristic it provides to their products. Starter culture rotation is not always possible, if the process involves the use of novel strains. The presence of bacteriophages in the environment leads to the quality instability of the product leads to the unexpected fermentation failure. In this context, a study of the prevalence of bacteriophages in the different sectors of dairy environment is carried out. The Screening of the phages has been undertaken with cheese and yoghurt cultures in the production and processing area. Three tests have been carried out to isolate the phages. Viz., turbidity test, double layer plaque assay, Spot test. Two natural temperate phages have been isolated from the cheese and yoghurt culture. Whereas the samples from the dairy processing environment is negative for phages. The temperate phages belong to the Siphoviridae family. Certain processing area where there is no involvement of starter culture e.g., paneer whey also showed the presence of phages in the whey, which indicates the air borne transmission of phages. Thus, the goal of this article is to make the quality assurance program associated with the production process of the fermented dairy products for the presence of phages in the dairy environment, with special emphasis on the preventive aspects related to this problem in the dairy fermentative industry.

Chironomus riparius is a well-established model organism in various fields such as ecotoxicology and ecology, and therefore environmental preferences, ecological interactions and metabolic traits are well-studied. With the recent publication of a high-quality draft genome, as well as different population genetic parameters such as mutation and recombination rate, the species can be used as an alternative to the Drosophila models in experimental population genomics or molecular ecology. To facilitate access to this promising experimental model species for a wider range of researchers, we describe experimental methods to first create and sustain long term cultures of C. riparius and then use them to perform repeatable and comparable experiments for various research questions.

To understand microbial growth with mathematical models has a long tradition that dates back to the pioneering work of Jacques Monod in the 1940s. Growth laws are simple mathematical expressions that aim at describing growth rates of microbes as functions of external parameters, in particular nutrient concentrations. These laws are now widely applied to construct, e.g., dynamic ecosystem models. However, to explain the growth laws from underlying (first) principles is extremely challenging. In the second half of the 20th century, numerous experimental approaches aimed at precisely measuring heat production during microbial growth to determine the entropy balance in a growing cell and to quantify the exported entropy. This has led to the development of thermodynamic theories of microbial growth, which have generated fundamental understanding and identified principle limitations of the growth process. Whereas these approaches considered a growing microbe as a black box, modern theories heavily rely on genomic resources to describe and model genome-scale networks to explain microbial growth. Interestingly, however, thermodynamic constraints are often included in modern modelling approaches only in a rather superficial fashion, and it appears that recent modelling approaches and classical theories are disconnected fields. In order to stimulate a closer interaction between these fields, we here review various theoretical approaches that aim at describing microbial growth based on thermodynamic principles. We start with classical black-box models of cellular growth, and continue with genome-scale modelling approaches that include thermodynamics, before we place these models in the context of fundamental considerations based on non-equilibrium statistical mechanics. We conclude by identifying conceptual overlaps between the fields and suggest how the various types of theories and models can be integrated. We outline how concepts from one approach may help to inform or constrain another, and we demonstrate how genome-scale models can be used to infer classical black-box parameters, which are experimentally accessible in growth experiments. Such integration will allow understanding to what extent microbes can be viewed as thermodynamic machines, and how close they operate to theoretical optima.

This study investigates distillery wastewater, commonly known as vinasse, as a potential culture medium for the production of Chlorella vulgaris and its most relevant metabolites. The effect of vinasse concentration on the composition of the biomass (proteins, carbohydrates and lipids) was evaluated in treatments performed in 6-L tubular air-lift reactors. The reactors were operated at 25 °C for 18 days, in total darkness, under a continuous flow of air. Results showed a rapid growth of microalgae in the first ten days, when an average production of 0.87 g/L was reached. Then, the daily biomass productivity began to decrease, up to an average value of 11.8 g/L at the 16th day. For all treatments, there was a significant reduction in the concentration of most metabolites in the first eight days. This was likely due to the adaptation of the biomass to the new conditions, with a transition from autotrophic to heterotrophic metabolism. From the 10th day, the concentration of metabolites in the biomass began to increase, reaching a nearly constant value at the 16th day. The observed maximum concentrations (%w/w) were: 48.95% proteins, 2.88% xylose, 7.82% glucose, 4.54% arabinose, 8.28% fructose and 4.82% lipids. These values were only marginally affected by the type of treatment. Overall, the results obtained suggest that vinasse is a promising and sustainable medium for the growth of C. vulgaris and the production of valuable metabolites.

This study investigates distillery wastewater, commonly known as vinasse, as a potential culture medium for the production of Chlorella vulgaris and its most relevant metabolites. The effect of vinasse concentration on the composition of the biomass (proteins, carbohydrates and lipids) was evaluated in treatments performed in 6-L tubular air-lift reactors. The reactors were operated at 25 °C for 18 days, in total darkness, under a continuous flow of air. Results showed a rapid growth of microalgae in the first ten days, when an average production of 0.87 g/L was reached. Then, the daily biomass productivity began to decrease, up to an average value of 11.8 g/L at the 16th day. For all treatments, there was a significant reduction in the concentration of most metabolites in the first eight days. This was likely due to the adaptation of the biomass to the new conditions, with a the transition from autotrophic to heterotrophic metabolism. From the 10th day, the concentration of metabolites in the biomass began to increase, reaching a nearly constant value at the 16th day. The observed maximum concentrations (%w/w) were: 48.95% proteins, 2.88% xylose, 7.82% glucose, 4.54% arabinose, 8.28% fructose and 4.82% lipids. These values were only marginally affected by the type of treatment. Overall, the results obtained suggest that vinasse is a promising and sustainable medium for the growth of C. vulgaris and the production of valuable metabolites.

Nowadays, the transparency of governments with respect to the sustainability of public services is a very interesting issue for stakeholders and academics. It has led to previous research and international organisations (EU, IMF, OECD, United Nations, IFAC, G-20, World Bank) to recommend promotion of the online dissemination of economic, social and environmental information. Based on previous studies about e-government and the influence of administrative cultures on governmental accountability, this paper seeks to identify political actions useful to improve the practices of transparency on economic, social and environmental sustainability in European local governments. We perform a comparative analysis of sustainability information published on the websites of 72 local governments in 10 European countries grouped into main three cultural contexts (Anglo-Saxon, Southern European and Nordic). Using international sustainability reporting guidelines, our results reveal significant differences in local government transparency in each context. The most transparent local governments are the Anglo-Saxon ones, followed by Southern European and Nordic governments. Based on individualized empirical results for each administrative style, our conclusions propose useful policy interventions to enhance sustainability transparency within each cultural tradition, such as development of legal rules on transparency and sustainability, tools to motivate local managers for online diffusion of sustainability information and analysis of information needs of stakeholders.

Stevia rebaudiana (Bert.) is a natural sweetener with diterpenic glycosides, offering 300 times more sweetness than sucrose, and boasting low-calorie content, along with medicinal and therapeutic properties. To address a limited genetic diversity, a germplasm bank comprising 115 segregated lines of stevia was established in Colombia. Notwithstanding, the fungus Athelia rolfsii adversely impacted key stevia lines, leading to wilting and plant mortality. For this reason our study aimed to explore the biocontrol potential of native strains of Trichoderma spp., isolated from productive lots in Colombia's Cordoba department, as a solution to manage A. rolfsii. Soil samples underwent serial dilution technique (103 conidia.mL-1) to obtain Trichoderma spp. isolates, which were thoroughly morphologically and molecularly characterized. A total five native strains were identified through sequencing the PCR products using universal ITS primers, and comparisons with the GENBANK database confirmed their identity (OK310695.1, ON237703.1, ON238106.1, ON237737.1, and ON237923.1). Their biocontrol efficacy against A. rolfsii was evaluated using the dual culture technique. Remarkably, all strains exhibited significant biocontrol capacity against the A. rolfsii isolate INVEPAR-05 (OK271308.1), except for Trichoderma viride INVEPAR-T10 (ON237923.1). These findings suggest that Trichoderma spp. could be a potential option to manage the fungus in stevia.

In this work, the resistance/susceptibility (R/S) profile of Staphylococcus equorum strains (n=30) from cheese to 16 antibiotics was determined by broth microdilution. The minimum inhibitory concentration (MIC) for all antibiotics was low in most strains, although higher MICs compatible with acquired genes were also noted. Genome analysis of 13 strains showed the S. equorum resistome to be composed of intrinsic mechanisms, acquired mutations, and acquired genes. However, the genetic data did not always correlate with the phenotype. As such, a cat gene providing resistance to chloramphenicol was found on a plasmid in one strain; this was able to provide resistance to Staphylococcus aureus after electroporation. An msr(A) polymorphic gene was identified in five strains. The Mrs(A) variants were associated with variable resistance to erythromycin. All strains harboured a polymorphic fosB/fosD gene, although only one acquired copy was associated with strong resistance to fosfomycin. Similarly, a plasmid-associated blaR1-blaZI operon encoding a penicillinase was identified in five ampicillin- and penicillin G-susceptible strains. Identified genes not associated with resistance further included mph(C) in two strains and norA in all strains. The antibiotic R/S status and gene content of S. equorum strains intended to be employed in food systems should be carefully determined.

The General Data Protection Regulation (GDPR) protects the personal data of natural persons and at the same time allows the free movement of such data within the European Union (EU). Hailed as majestic by admirers and dismissed as protectionist by critics, the Regulation is expected to have a profound impact around the world, including in the African Union (AU). For European–African consortia conducting research that may affect the privacy of African citizens, the question is ‘how to protect personal data of data subjects while at the same time ensuring a just distribution of the benefits of a global digital ecosystem?’ We use location privacy as a point of departure, because information about an individual’s location is different from other kinds of personally identifiable information. We analyse privacy at two levels, individual and cultural. Our perspective is interdisciplinary: we draw from computer science to describe three scenarios of transformation of volunteered/observed information to inferred information about a natural person and from cultural theory to distinguish four privacy cultures emerging within the EU in the wake of GDPR. We highlight recent data protection legislation in the AU and discuss factors that may accelerate or inhibit the alignment of data protection legislation in the AU with the GDPR.

Brucellosis is one of the most common etiologies of laboratory-acquired infections worldwide, and handling of living brucellae should be performed in a Class II biological safety cabinet. The low infecting dose, multiple portals of entry to the body, the great variety of potentially contaminated specimens, and the unspecific clinical manifestations of human infections facilitate the unintentional transmission of brucellae to laboratory personnel. Work accidents such as spillage of culture media cause only a small minority of exposures, whereas >80% of events result from unfamiliarity with the phenotypic features of the genus, misidentification of isolates, and unsafe laboratory practices such as aerosolization of bacteria and working on an open bench without protective goggles or gloves. Although the bacteriological diagnosis of brucellae by traditional methods is simple, the Gram stain and the biochemical profile of the organism, as determined by commercial kits, can be misleading, resulting in inadvertent exposure and contagion. The use of novel identification technologies is not hazard-free. The MALDI-TOF technology requires an initial bacterial inactivation step, while the instruments’ reference database may misidentify Brucella as belonging to other Gram-negative species. The rapid identification by the FISH method mistakes brucellar isolates for members of the closely related Ochrobactrum genus.

Tea plants (Camellia sinensis L.) are economically important horticultural crops for industrial use. The drink (tea) obtained from them is used all over the world, and its functional activity is determined by the polyphenols content and composition. Its accumulation is regulated by the action of many environmental factors. This paper presents data on the different intensity (50, 75, and 100 µmol·m−2·s−1) and duration of light exposure on tea plant in vitro cultures. Changes in morphophysiological characteristics, accumulation of carbohydrates and polyphenols, including their monomeric (phenylpropanoids, flavanols) and oligomeric (proanthocyanidins) forms, were studied. The malondialdehyde and photosynthetic pigments (chlorophyll a and b) contents were also analyzed. In the calli grown in light conditions, chloroplasts and anthocyanoplasts formation was noted, which was especially pronounced under the highest illumination intensity. A significant similarity between the growth of callus cultures, the carbohydrates and polyphenols accumulation at 50 and 75 µmol·m−2·s−1 was established. Which wasn’t characteristic of the calli grown at 100 µmol·m−2·s−1. The stress reaction absence in tea callus cultures has been demonstrated, which may be a consequence of the phenolic antioxidants’ accumulation in them. The obtained data evidence that their potential ability to accumulate primary (carbohydrates) and secondary (polyphenols) metabolites depends on the light exposure intensity and duration.

(1) Background: Central nervous system (CNS) development is characterized by dynamic changes in cell proliferation and differentiation. Key regulators of these transitions are the transcription factors such as SOX2 and SOX9. SOX2 is involved in the maintenance of progenitor cell state and neural stem cell multipotency, while SOX9, expressed in neurogenic niches, plays an important role in neuron/glia switch with predominant expression in astrocytes in the adult brain. (2) Methods: To validate SOX2 and SOX9 expression pattern in dissociated primary neuronal cultures derived from opossum (Monodelphis domestica), we used immunohistochemistry (IHC) and the isotropic fractionator method on fixed cortical tissue from comparable postnatal ages; (3) Results: Neurons positive for both neuronal (NeuN or TUJ1) and stem cell (SOX2) markers were identified and their presence was confirmed with all methods and postnatal age groups (P4-6, P6-18 and P30) analyzed. SOX9 showed exclusive staining in non-neuronal cells, and it was coexpressed with SOX2. (4) Conclusions: The persistence of SOX2 expression in developing cortical neurons of M. domestica during first post-natal month implies the functional role of SOX2 during neuronal differentiation and maturation, which was not previously reported in opossums.

The production and N-glycosylation of recombinant human butyrylcholinesterase (BChE), a model highly glycosylated therapeutic protein, in a transgenic rice cell suspension culture treated with kifunensine, a strong α-mannosidase I inhibitor, was studied in a 5 L bioreactor. A media exchange was performed at day 7 of cultivation by removing spent sugar rich media (NB+S) and adding fresh sugar free (NB-S) media to induce the rice α-amylase 3D (RAmy3D) promoter to produce rice recombinant human BChE (rrBChE). Using a 1.25X concentrated sugar-free medium together with an 80% reduced working volume during the media exchange lead to a total active rrBChE production level of 79 ± 2 µg (g FW)^-1 or 7.5 ± 0.4 mg L^-1 in the presence of kifunensine, which is 1.5-times higher than our previous bioreactor runs using normal sugar free medium with no kifunensine treatment. Importantly, the amount of secreted active rrBChE in culture medium was enhanced in the presence of kifunensine, comprising 44% of the total active rrBChE at day 5 post-induction. Coomassie stained SDS-PAGE gel and Western blot analyses reveal different electrophoretic migration of purified rrBChE bands with and without kifunensine treatment, which is attributed to different N-glycoforms. N-Glycosylation analysis shows substantial increase of oligomannose glycans (Man5/6/7/8) in rrBChE treated with kifunensine compared to controls. However, mass transfer limitation of kifunensine is likely the major reason for incomplete inhibition of α-mannosidase I in this bioreactor study.

During meat processing, lactic acid bacteria (LAB) have to competitively adapt to the hostile en-vironment produced by curing additives (CA). The objective of this study was to investigate the ability of Latilactobacillus curvatus CRL 705, a bioprotective strain of meat origin, to adapt to CA. A physiological and proteomic approach was performed. CRL 705 was grown in a chemically de-fined medium (CDM) containing specific concentrations of CA (NaCl, nitrite, sucrose and ascor-bic acid). The results showed minor differences in growth kinetics in the presence of CA. Glucose consumption, present in CDM, and production of lactic acid and bacteriocins were not signifi-cantly affected. Proteomic analyses indicated that most of the identified proteins (36 out of 39) mainly related to carbohydrate metabolism (18%), posttranslational modifications (15.6%), energy production and conversion (11.1%), translation (11.1%) and nucleotide metabolism (8.9%), were under expressed. In response to the studied CA, CRL 705 slowed down its general metabolism, achieving slight changes in physiological and proteomic parameters. The observed performance is another characteristic that extends the well-known competitive profile of CRL 705 as a meat starter- and -bioprotective culture. This is the first report dealing with the impact of CA on LAB proteomics.

Rural tourism plays an increasing role in maintaining sustainable rural development. In-11 tegrating culture into rural tourism is multifaceted. Local communities have often been regarded as 12 homogeneous, and different voices within them are selectively presented or re-interpreted by pow-13 ers. A better understanding of how and why cultures are integrated into rural tourism is urgently 14 needed. This paper aims to investigate (1) the aims and motives of tourism managers to integrate 15 cultural concepts into rural tourism; (2) who has participated in the cultural integration process; and 16 (3) how cultures have been integrated into rural tourism. Based on a scoping literature review, we 17 found that cultures could add more attractions, such as historical heritage, artwork, cultural land-18 scape, customs, food, and language, to the natural landscape and bring more tourists to rural areas. 19 However, integrating cultures into rural tourism is not always successful. Conflicts of interest 20 among different stakeholders are also often found. Some of the worst cases of cultural integration 21 have even destroyed the original natural landscape and local cultures. While cultural integration is 22 complicated, injustice and perverse effects do not have to be a consequence of integrating cultures 23 into rural tourism. Multi-way communication among tourism providers, managers, and consumers 24 can mitigate disruptive outcomes and unlock positive social outcomes.

Gamma-aminobutyric acid (GABA) is thought to have beneficial effects on human health and well-being. GABA enrichment by lactic acid bacteria (LAB) could be used in the development of functional fermented foods. In this work, four GABA-producing strains each of Lactococcus lactis and Streptococcus thermophilus were isolated from cowʼs milk, and their phenotypic, technological, and safety profiles were determined. Genome analysis provided genetic support for the majority of the analyzed traits. The operon harboring the glutamate decarboxylase gene (gadB) was chromosomally encoded in all strains and showed the same gene content and gene order as that reported, respectively, for L. lactis and S. thermophilus. In the latter species, the operon was flanked (as in most strains of this species) by complete or truncated copies of insertion sequences (IS), suggesting recent acquisition by horizontal gene transfer. The genomes of three L. lactis and two S. thermophilus strains showed a gene encoding a caseinolytic proteinase (PrtP in L. lactis and PrtS in S. thermophilus). These strains grew in milk forming a coagulum of good appearance and an appealing acidic flavor and taste. They also produced GABA from monosodium glutamate in this medium. Two L. lactis strains were identified as belonging to the biovar. diacetylactis, utilized citrate from milk and produced significant amounts of acetoin. None of the strains showed any noticeable antibiotic resistance, nor did their genomes harbor transferable antibiotic resistance genes, genes involved in toxicity, virulence, or pathogenicity. Altogether these results suggest all eight strains may be considered candidates for use as starter or components of mixed LAB cultures for the manufacture of GABA-enriched fermented dairy products.

The growth of two pairs of co-cultures (Escherichia coli/Geotrichum candidum, and Staphylococcus aureus/Geotrichum candidum) with a starter culture of lactic acid bacteria was studied in milk at temperatures related to artisanal cheesemaking ripening. For an inoculum of approximately 106 CFU/mL, LAB not only induced an early stationary phase of E. coli (two strains BR and PS2) and S. aureus (strains 2064 and 14733) but affected their death phase. G. candidum was found to be the subject of interactions with LAB within a given temperature range only partially. To develop a tertiary model for the growth curves of the populations, a one-step approach was used, combining two types of primary models (Huang and Gimenez and Dalgaard) with secondary square root models for growth rate and lag time. Furthermore, the reparametrized Gompertz-inspired function with the Bigelow secondary model was used to describe the death phase of the E. coli and S. aureus strains. The prediction ability of the growth of the H-GD tertiary model for co-cultures was cross-validated within the strains and data sets in milk and milk medium with 1 % NaCl.

The mammalian gut ecosystem plays critical roles in multiple functions related to health and homeostasis. In many cases, disturbances in the gut ecosystem are associated with a large number of metabolic and chronic diseases and disorders such as diabetes, cancer, and obesity. A diverse community of microorganisms ranging from viruses to bacteria comprise the gut microbiota, which is often considered as an organ in itself. Recent studies have profiled the influence of lifestyles and dietary behavior by comparing the gut microbiome of populations with different cultural underpinnings. In this review, we provide an overview of the studies which report the influence on the gut microbial composition of dietary and lifestyle patterns in different contexts such as western industrialized countries and indigenous cultures (corresponding to different lifestyle gradients such as hunter-gatherers and pastoralists) and how this association may influence health and disease.

Arthrospira platensis, commercially known as Spirulina, is a fresh-water cyanobacterium that is gaining even more attention in the last years due to its high biological and nutritional value. For this reason, it has been employed in several food applications, to obtain or enhance functional and technological properties of cheese, yogurt, bread, cookies or pasta. The aim of this work was to evaluate the potential boosting effect of two different concentrations (0.25% and 0.50%) of Arthrospira platensis on the fermentation capability of several starter LAB strains, 1 probiotic and 4 commercial mix culture. These strains were used to ferment three different substrates and their fermentation behaviors were evaluated by impedance analyses together with rheological and color measurements. It was demonstrated that the booster effect took place, but it was variable and dependent not only on the strain or mix culture used, but also on the substrate and Arthrospira platensis concentration. Also, rheological and color modifications were found to be dependent of these factors.

Nine 2-(1,2-benzothiazol-3-yl)-N-(4-oxo-2-phenyl-1,3-thiazolidin-3-yl) propanamides were designed and synthesized, combining benzisothiazole and 4-thiazolidinone in one frame. The aim of the study was to verify their effectiveness to contrast the inflammatory/oxidative process in which free oxygen and nitrite (ROS and RNS) radicals, advanced glycation products (AGEs), inflammatory cytokines and matrix metalloproteinases (MMPs) are involved. Docking studies of all the compounds were performed in order to explore their binding mode at the MMP-9 protein. An appreciable anti-inflammatory/wound healing effects of the tested compounds was highlighted. Derivative 23, bearing a 4-carboxyphenyl substituent at C2 of the 4-thiazolidinone ring, exhibited the highest activity, being able to inhibit MMP-9 at nanomolar level (IC₅₀ = 40 nM).

Gymnema sylvestre is a pharmacological plant which has a rich source of bioactive compounds specifically gymnemic acid (GA) and phenolic compounds (PC) that used for pharmaceutical industries. Sources for naturally occurring bioactive compounds are limited, due to geographical and seasonal variations; on the other hand, it is commercially in demand. Biosynthesis of G. sylvestre phytochemicals through in vitro culture often enhanced by elicitation. The use of cell suspension cultures (CSC) has interested serious attention on the production of essential phytochemicals. The current study is aimed at improving the contents of GA and PC in G. sylvestre CSC using the copper oxide nanoparticles (CuO NPs). Callus was obtained on MS medium with 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), 0.1 mg/L kinetin (KIN), phytoagar (8.0 g/L), and sucrose (30 g/L). The above medium devoid of agar was used for the initiation of CSC. The CSC was treated with three levels of CuO NPs (1, 3 or 5 mg/L) to enhance the production of GA and PC. The greatest amount of GA (89.25 mg/g dry cell mass, DCM), total phenolic (245.10 mg/g), and flavonoid (4.57 mg/g) in CSC were achieved when G. sylvestre cells were treated for 48 h with 3 mg/L CuO NPs. Also, the biomedical potential (antioxidant, antidiabetic, anti-inflammatory, antibacterial, antifungal and anticancer activities) were also high in the CuO NPs (3 mg/L) treated CSC extracts of G. sylvestre. CuO NPs elicitation of CSC significantly increased production of GA (9-fold), and PC than non-elicited CSC in G. sylvestre.

With worsening environmental degradation and increasing international demand for human health, plant cell based technology will be more important. The aim of this review is to give an overview regarding key points of current progress and development trend of nature medical and edible plants from the framework of whole plant cell engineering including cell cultures, cell fusion and cell splitting derived biotechnology. Here we sum up three main application trends of medical plants cell cultures covering industrial output, technology upgrading and potential resource development. We also provide classification and typification insights to summarize the development of somatic hybridization in cereal, vegetables crops and other medicinal and edible plants as well as application of artificial polyploidy induction in major medicinal herbs. Development medicinal and edible homologous plants based on modern plant cell engineering will bring us a promising future.

Chronic degeneration of the Retinal Pigment Epithelium (RPE) is a precursor to pathological changes in the outer retina. The RPE monolayer, which lies beneath the neuroretina, daily internalises and digests large volumes of spent photoreceptor outer segments. Impaired cargo handling and processing in the endocytic/phagosome and autophagy pathways leads to the accumulation of lipofuscin and N-retinylidene-N-retinylethanolamine aggregates and chemically-modified compounds such as malondialdehyde and 4-hydroxynonenal within RPE. These contribute to increased proteolytic and oxidative stress, resulting in irreversible damage to post-mitotic RPE cells and development of blinding conditions such as Age-related Macular Degeneration, Stargardt disease and Choroideremia. Here, we review how impaired cargo handling in the RPE results in their dysfunction, discuss new findings from our laboratory and consider how newly discovered roles for lysosomes and the autophagy pathway could provide insights into retinopathies. Studies of these dynamic, molecular events have also been spurred on by recent advances in optics and imaging technology. Mechanisms underpinning lysosomal impairment in other degenerative conditions including storage disorders, a-synuclein pathologies and Alzheimer’s disease are also discussed. Collectively, these findings help transcend conventional understanding of these intracellular compartments as simple waste disposal bags to bring about a paradigm shift in the way lysosomes are perceived.

Bacterial Calcium-carbonate Precipitation (BCP) has been studied for multiple applications such as remediation, consolidation and cementation. Isolation and screening of strong calcifying bacteria is the main task of BCP-technique. In this paper we studied CaCO₃ precipitation by different bacteria isolated from a rhizospheric soil in both solid and liquid media. It has been found, through culture-depending studies, that bacteria belonging to Actinobacteria, Gammaproteobacteria and Alphaproteobacteria are the dominant bacteria involved in CaCO₃ precipitation in this environment. Pure and mixed cultures of selected strains were applied for sand biocementation experiments. Scanning Electron Microscopy (SEM) and Energy Dispersive X-ray Spectroscopy (EDS) analyses of the biotreated samples revealed the biological nature of the cementation and the effectiveness of the biodeposition treatment by mixed cultures. X-ray diffraction (XRD) analysis confirmed that all the calcifying strains selected for sand biocementation precipitated CaCO₃, mostly in the form of calcite. In this study Biolog® Eco-plate is evaluated as a useful method for a more targeted choice of the sampling site with the purpose of obtaining interesting candidates for BCP applications. Furthermore, ImageJ software was investigated, for the first time to our knowledge, as a potential method to screen high CaCO₃ producer strains.

HIV-associated neurocognitive disorders (HAND) persist under antiretroviral therapy as a complex pathology that has been difficult to study in cellular and animal models. Therefore, we generated an ex vivo human brain slice model of HIV-1 infection from surgically resected adult brain tissue. Brain slice cultures processed for flow cytometry showed >90% viability of dissoci-ated cells within the first three weeks in vitro, with parallel detection of astrocyte, myeloid, and neuronal populations. Neurons within brain slices showed stable dendritic spine density and mature spine morphologies in the first weeks in culture, and they generated detectable activity in multi-electrode arrays. We infected cultured brain slices using patient-matched CD4+ T-cells or monocyte-derived macrophages (MDMs) that were exposed to a GFP-expressing R5-tropic HIV-1 in vitro. Infected slice cultures expressed viral RNA and developed a spreading infection up to 9-days post-infection, which were significantly decreased by antiretrovirals. We also detected infected myeloid cells and astrocytes within slices and observed minimal effect on cellular via-bility over time. Overall, this human-centered model offers a promising resource to study the cel-lular mechanisms contributing to HAND (including antiretroviral toxicity, substance use, and aging), infection of resident brain cells, and new neuroprotective therapeutics.

Clear cell renal cell carcinoma (ccRCC) is the most prevalent form of renal cancer and its treatment is hindered by resistance to targeted and immunotherapies and combinations ob both. We have reported that knockdown of the antisense noncoding mitochondrial RNAs (ASncmtRNAs) with chemically-modified antisense oligonucleotides induces proliferative arrest and apoptotic death in tumor cells from many human and mouse cancer types. These studies have been mostly performed in vitro and in vivo on commercially-available cancer cell lines and have shown that, in mouse models, tumor growth is stunted by the treatment. The present work was performed on cells derived from primary and metastatic ccRCC tumors. We established primary cultures fom primary and metastatic ccRCC tumors, which were subjected to knockdown of ASncmtRNAs in vitro and in vivo in an orthotopic xenograft model in NOD/SCID mice. We found that these primary ccRCC cells are affected in the same way as tumor cell lines and, in the orthotopic model, tumor growth was significantly reduced by the treatment. This study on patient-derived ccRCC tumor cells represents a model closer to actual patient ccRCC tumors and shows that knockdown of ASncmtRNAs poses a potential treatment option for these patients.

Thai basil is a renowned medicinal plant and a rich source of bioactive antioxidant compounds having several health benefits, with actions to prevent of cancer, diabetes and cardiovascular disease. Plant cell and tissue culture technologies can be routinely established as an important, sustainable and low-cost biomass source for the production of high-value phytochemicals. The current study aimed at developing an effective protocol for the production of Thai basil leaf derived callus cultures with sustainable and high production of biomass and antioxidants as an alternative of leaves production. MS basal medium with various concentrations of plant growth regulators (PGRs) compatible with nutraceutical applications (i.e., gibberellic acid (GA3) and 6-benzylaminopurine (BAP) either alone or in combination with naphthalene acetic acid (NAA)) were evaluated. Among all tested PGRs, the combination BAP:NAA (5 mg/L:1 mg/L) yield maximum biomass accumulation (fresh weight (FW): 190 g/L and dry weight (DW): 13.05 g/L) as well as enhanced phenolic (346.08 mg/L) production. HPLC quantification analysis indicated high productions of chicoric acid (35.77 mg/g DW) and rosmarinic acid (7.35 mg/g DW) under optimized callus culture conditions. Antioxidant potential was assessed using both in vitro cell free and in vivo cellular antioxidant assays. Maximum in vitro antioxidant activity DPPH (93.2 % of radical scavenging activity) and ABTS (1322 µM Trolox equivalent antioxidant capacity) was also observed for the extracts from callus cultures grown on optimal conditions. In vivo cellular antioxidant activity assay confirmed the effective protection against oxidative stress of the corresponding extract by the maximum inhibition of ROS and RNS production. Compared to commercial leaves, callus extracts showed higher production of chicoric acid and rosmarinic acid associated with higher antioxidant capacity. In addition, this biological system also has a large capacity for continuous biomass production, thus demonstrating its high potential for possible nutraceutical applications.

There is a huge interest in developing novel hollow fiber (HF) membranes able to modulate neural differentiation to produce in vitro blood-brain barrier (BBB) models for biomedical and pharmaceutical research, due to the low cell-inductive properties of the polymer HFs used in current BBB models. In this work, poly(ε-caprolactone) (PCL) and composite PCL/graphene (PCL/G) HF membranes were prepared by phase inversion and were characterized in terms of mechanical, electrical, morphological, chemical, and mass transport properties. The presence of graphene in PCL/G membranes enlarged the pore size and the water flux and presented significantly higher electrical conductivity than PCL HFs. Biocompatibility assay showed that PCL/G HFs significantly increased C6 cells adhesion and differentiation towards astrocytes, may be attributed to their higher electrical conductivity in comparison to PCL HFs. On the other hand, PCL/G membranes produced a cytotoxic effect on the endothelial cell line HUVEC presumably related with a higher production of intracellular reactive oxygen species induced by the nanomaterial in this particular cell line. These results prove the potential of PCL HF membranes to grow endothelial cells and PCL/G HF membranes to differentiate astrocytes, the two characteristic cell types that could develop in vitro BBB models in future 3D co-culture systems.

Traditional dry fermented meat products are obtained artisanally in many countries, where they represent a gastronomic heritage well distinguished from industrial counterparts. This food category is most often obtained from red meat, a food commodity that is under attack because of evidences of increased risk of cancer and degenerative diseases with high consumption. However, traditional fermented meat products are intended for moderate consumption and gastronomic experience and, as such, their production must be continued also to safeguard culture and economy of the geographical areas of origin. In this review, the main risks attributed to these products are considered and how these risks are minimized by following precise production norms, respectful of the ancient manufacturing processes is described. Moreover, results obtained in studies reporting use of autochthonous microbial cultures of lactic acid bacteria (LAB), coagulase negative staphylococci (CNS), Debaryomyces hansenii and Penicillium nalgiovense to improve safety and quality of traditional fermented meats are summarized. The role of these products as a source of microorganisms that can be beneficial to the host are also considered. From the results of the studies reviewed here it appears that the development of autochthonous cultures for these foods ensure safety and stabilize sensory characteristics with possibility to be extended to the large variety of traditional productions.

Thirteen benzylethylenearyl ureas and one carbamate have been designed, synthesized and biologically evaluated as regards their antiproliferative activity on tumor cell lines HEK-293 and A-549 and on the immune and endothelial Jurkat T cells and HMEC-1, respectively. A relationship between the structure of the synthetic compounds and their antiproliferative activity has been established. Compounds C.1, C.3, C.12 and C.14 were chosen for further biological studies in order to determine their potential as oncoimmunomodulating agents. Some compounds exhibited significant inhibitory effects on both total PD-L1 and VEGFR-2 in HT-29 cell line, showing urea C.12 active against both targets. Some compounds could reduce more than 50% of cancer living cells, compared to non-treated ones, when assessed in co-cultures using HT-29 and THP-1 cells. In addition, these compounds showed no effect on the immune cell viability and significantly reduce CD11b expression, which is a promising target for immune modulation in anti-cancer therapies.