Production of Stilbenes in Callus Cultures of the Maltese Indigenous Grapevine Variety, Gellewza.

The production of secondary metabolites in tissue culture has been considered as an alternative to the cultivation and harvesting of crops intended for this purpose. The present study was aimed at the growth of callus and production of polyphenolic compound of callus derived from a Maltese indigenous grapevine variety, Gellewza. Callus was inoculated onto plant growth regulators-enriched Murashige Skoog media (MSm) to determine whether polyphenols are produced in vitro as well as to determine the best combination of plant growth regulators needed for the production of these metabolites. From results obtained, it was observed that the best callus production was obtained by auxin-enriched MSm. In fact, indole acetic acid and indole acetic acid /6-benzyl aminopurine enhanced biomass accumulation (3.04 g and 3.39 g) as opposed to the others (<1.97 g). On the other hand, parameters showing the presence of flavonoids (tonality, 3.80), particularly anthocyanins (24.09 mg/kg) and total polyphenols (1.42 mg/g), were optimum in the presence of cytokinins, particularly 6-benzyl aminopurine. Analysis for single polyphenols revealed a high amount a particular stilbene: polydatin (glucoside of resveratrol). Resveratrol and other typical polyphenols, found in mature berries, were also found in significant quantities, while the other polyphenolic compounds were found in minimal quantities. This is the first study to describe the production and composition of polyphenols in Gellewza callus cultures. From the results obtained, it can be seen that this grape tissue is an excellent alternative for the production of polyphenols from the stilbene group, which can be upscaled and exploited commercially.

Oleuropein Prevents Neuronal Death, Mitigates Mitochondrial Superoxide Production and Modulates Autophagy in a Dopaminergic Cellular Model.

Parkinson's disease (PD) is a progressive neurodegenerative disorder, primarily affecting dopaminergic neurons in the substantia nigra. There is currently no cure for PD and present medications aim to alleviate clinical symptoms, thus prevention remains the ideal strategy to reduce the prevalence of this disease. The goal of this study was to investigate whether oleuropein (OLE), the major phenolic compound in olive derivatives, may prevent neuronal degeneration in a cellular dopaminergic model of PD, differentiated PC12 cells exposed to the potent parkinsonian toxin 6-hydroxydopamine (6-OHDA). We also investigated OLE's ability to mitigate mitochondrial oxidative stress and modulate the autophagic flux. Our results obtained by measuring cytotoxicity and apoptotic events demonstrate that OLE significantly decreases neuronal death. OLE could also reduce mitochondrial production of reactive oxygen species resulting from blocking superoxide dismutase activity. Moreover, quantification of autophagic and acidic vesicles in the cytoplasm alongside expression of specific autophagic markers uncovered a regulatory role for OLE against autophagic flux impairment induced by bafilomycin A1. Altogether, our results define OLE as a neuroprotective, anti-oxidative and autophagy-regulating molecule, in a neuronal dopaminergic cellular model.

Chemical Characterization and In Vitro Gas Production Kinetics of Alternative Feed Resources for Small Ruminants in the Maltese Islands.

The ever-increasing human population, the problem associated with climate change and recent crises-COVID-19 disease and trade conflicts-all impacted on the availability and cost of animal feed raw materials. This is clearly visible in realities which heavily rely on importation such as islands and small states, where producers involved in the agricultural sector were strongly affected by the sharp increase in prices. To deal with these global issues, alternative resources are perceived to replace conventional ingredients. This work aimed at assessing the nutritive value of different resources (sheep feed, mature carob, Maltese bread, wild asparagus, prickly lettuce, and loquat) for small ruminants present in the Maltese Islands, analyzing their chemical composition, gas production kinetics and antioxidant properties. In general, the variation in chemical composition resulted in different rumen fermentation kinetics (p < 0.007). The ratio between GP-24 h and GP-48 h was higher in Maltese bread than other substrates; loquat, prickly lettuce and wild asparagus showed lower fermentation kinetics in accordance with their high NDF and ADF contents. The antioxidant activity may be partially related to the polyphenolic content that was higher in wild asparagus, prickly lettuce and loquat. All feed characteristic confirmed their potential to be included as ingredients in ruminant diets and as a source of fiber.

Opuntia spp. as Alternative Fodder for Sustainable Livestock Production.

During the past decades, livestock production has increased significantly, which has led to the degradation of rangelands due to overgrazing. The lack of water in several arid areas has led to a decline in crop and animal husbandry. As a consequence, the demand for drought-resistant crops has increased significantly so as to keep crop and animal husbandry systems viable and sustainable. Cactaceae have adaptive characteristics that ensure their development progress under drought conditions. The present review provides information on the nutritive value of Opuntia in animal fodder production, its effects on animal performance, and the quality of the animal-derived products. In conclusion, the use of Opuntia as innovative alternative feed would render animal production systems more sustainable.

Selection of Acetic Acid Bacterial Strains and Vinegar Production From Local Maltese Food Sources.

This study investigates the isolation, identification, and fermentation performance of autochthonous acetic acid bacteria (AAB) from local niche habitats on the Island of Gozo (Malta) and their further use for vinegar production, employing local raw materials. The bacteria were isolated from grapevine berries and vinegar produced in the cottage industry. Following phenotype and genotype identification, the AAB were ascribed to the genera Acetobacter, Gluconobacter, and Komagataeibacter. A mixture of selected AAB was tested as an inoculum for vinegar production in bench fermenters, under different conditions and substrates, namely, grapes, honey, figs, onions, prickly pear, and tomatoes. The bench fermenters were operated under semi-continuous fermentation where working volumes were maintained by discharging and subsequent recharging accordingly to maintain the acidity in fermenters by adding 30-50 g/l of acetic acid for optimal Acetobacteraceae performance. Finally, the vinegar products obtained from the different substrates were evaluated for their quality, including organoleptic properties, which showed the superior quality of wood-treated vinegar samples with respect to neat vinegar samples.

Coffee intake during pregnancy and neonatal low birth weight: data from a multicenter Italian cross sectional study.

Aim: Coffee intake is common during pregnancy. However, the influence of coffee and caffeine on pregnancy has not yet been fully determined. Some studies show that high coffee intake could cause miscarriage, preterm birth or reduction of fetal growth, but other studies do not support these findings. The aim of the present study was to analyze data collected from a database focusing on coffee intake during pregnancy, which was specifically created for multicenter studies carried out in the maternity units of Italian general hospitals. Principal outcomes of pregnancy during pregnancy were considered. Methods: Data of 5405 pregnancies were collected by a direct questionnaire supplemented with data from patients'clinical records during the survey named PHYTO.VIG.GEST. Results: We observed that 42.3% of the total sample had consumed at least one coffee a day during pregnancy. Analysis of a dose-response relationship showed that, in pregnant women starting from the consumption of three coffees a day (6% of pregnant women consuming coffee), there is a statistically significant association between number of coffees and reduction of babies birth weight (< 2500 g). Coclusion: Even though high coffee intake is known to influence negatively birth weight, our results indicate that a significant percentage of pregnant women maintain this habit.

Consumption of Minerals, Toxic Metals and Hydroxymethylfurfural: Analysis of Infant Foods and Formulae.

Infant foods and formulae may contain toxic substances and elements which can be neo-formed contaminants or derived from raw materials or processing. The content of minerals, toxic elements, and hydroxymethylfurfural (HMF) in infant foods and formulae were evaluated. The effect of storage temperature on HMF formation in infant formulae and its potential as a quality parameter was also evaluated. Prune-based foods contained the highest HMF content. HMF significantly increased when the storage temperature was elevated to 30 °C for 21 days. All trace elements were present in adequate amounts, while the concentration of nickel was higher when compared to those of other studies. The study indicates that HMF can be used as a quality indicator for product shelf-life and that the concentrations of minerals and toxic elements vary greatly due to the diverse compositions of foods and formulae. Such contaminants need to be monitored as infants represent a vulnerable group compared to adults.

Cows fed hydroponic fodder and conventional diet: effects on milk quality.

The technology of green fodder production is especially important in arid and semiarid regions. Hydroponics improves on average the amount of crops in the same space, as traditional soil-based farming and can reduce water consumption compared to traditional farming methods. Limited research has been carried out on the use of hydroponic fodder and milk quality. A comparative study of traditional (Malta farm) and hydroponic fodder (Gozo farm) was conducted in Malta with 20 cows of the Holstein-Friesian breed from two farms. Individual and bulk-tank milk samples were collected once a week for a period of 1 month in order to evaluate physical (pH, conductivity, density, freezing point) and chemical (fat, protein, ash, lactose, solid nonfat) parameters as well as mineral (Zn, Cu, Pb, Ba) content. Milk proximate and physical data were processed by analysis of variance (ANOVA) for repeated measures and an ANOVA procedure with farm and time as effects for minerals. The results indicated differences in fat content and pH, showing higher values ( P < 0.05 ) in milk samples of cows fed with the hydroponic rather than the traditional fodder; a significant time effect ( P < 0.001 ) was found in all qualitative analyses except for lactose and salts. Minerals were in the range as reported elsewhere; Cu and Pb content was significantly higher ( P < 0.001 ) in the Gozo farm than the one in Malta, whereas Zn content showed higher values in Malta ( P < 0.001 ) than Gozo. Although the proximate results were similar for both farms, except for the higher fat content for the Gozo farm, principal component analysis (PCA) revealed that milk quality for the Gozo farm was superior to that of the Malta farm. However, further studies are needed to determine the effects of different hydroponic fodder using a large herd size.

Cucurbitacin E, An Experimental Lead Triterpenoid with Anticancer, Immunomodulatory and Novel Effects Against Degenerative Diseases. A Mini-Review.

A growing number of studies have revealed that natural molecules own interesting antioxidant and anti-apoptotic properties in cell culture as well as in animal models of human diseases such as cancer, inflammatory and neurodegenerative diseases. During the past sixty years, several cucurbitacins have been isolated from a number of cucurbitaceous species, amongst others. Cucurbitacins are triterpenoid compounds originally identify as the bitter components of the Cucurbit family that demonstrated several pro-survival activities in various model of cellular decay. Specifically, Cucurbitacin E (CuE), an oxygenated tetracyclic triterpenoid, has been investigated in a wider array of bioactivities, mainly immunomodulatory. Recently, CuE has been reported to possess anti-inflammatory and anti-tumorigenic properties mediated by its action on the cellular cytoskeleton, on mitotic pathways as well as on cellular autophagy. Few studies also pinpoint the role of CuE in the nervous system as cytostatic for gliomas and neuroprotective in a model of Parkinson's diseases. This review deals with the use of CuE in various experimental models as one of the most promising therapeutic natural molecules against cancer proliferation, as an immunomudulator and for the prevention of neurodegeneration.

In vitro fermentation of feed ingredients by fresh or frozen pig fecal inocula.

In vitro techniques can offer a rapid, repeatable and cost-effective alternative to in vivo experiments. We investigated the gas production (GP) of nine different feeds commonly used in Italian and Maltese pig farms in order to assess the use of the Ankom system in this species and to compare the in vitro microbial activity fresh or frozen inocula derived from piglet feces. Proximate and polyphenolic analyses were determined for all substrates. Fermentation profiles were measured according to the cumulative GP technique. Analysis of GP values revealed significant differences in the fermentation patterns mainly due to substrate and inoculum effects. Fermentation parameters varied significantly according to the substrate ingredients. The frozen inoculum generally led to more total GP than for fresh, with the exception of hard wheat bran pellets (HWBP), pelleted pulp and mature carob. Conversely the fresh fecal inoculum had an earlier maximum time to reach maximum GP than the frozen, showing a higher maximum rate of GP in all substrates excepting for HWBP. A significant difference in frozen and fresh inocula was found for the complete formulation. Further studies are required to confirm the variation between fresh and frozen fecal inocula to a wider range of feed, including those that could have negative biological effects.

Cucurbitacin E has neuroprotective properties and autophagic modulating activities on dopaminergic neurons.

Natural molecules are under intensive study for their potential as preventive and/or adjuvant therapies for neurodegenerative disorders such as Parkinson's disease (PD). We evaluated the neuroprotective potential of cucurbitacin E (CuE), a tetracyclic triterpenoid phytosterol extracted from the Ecballium elaterium (Cucurbitaceae), using a known cellular model of PD, NGF-differentiated PC12. In our postmitotic experimental paradigm, neuronal cells were treated with the parkinsonian toxin 1-methyl-4-phenylpyridinium (MPP(+)) to provoke significant cellular damage and apoptosis or with the potent N,N-diethyldithiocarbamate (DDC) to induce superoxide (O2(•-)) production, and CuE was administered prior to and during the neurotoxic treatment. We measured cellular death and reactive oxygen species to evaluate the antioxidant and antiapoptotic properties of CuE. In addition, we analyzed cellular macroautophagy, a bulk degradation process involving the lysosomal pathway. CuE showed neuroprotective effects on MPP(+)-induced cell death. However, CuE failed to rescue neuronal cells from oxidative stress induced by MPP(+) or DDC. Microscopy and western blot data show an intriguing involvement of CuE in maintaining lysosomal distribution and decreasing autophagy flux. Altogether, these data indicate that CuE decreases neuronal death and autophagic flux in a postmitotic cellular model of PD.

Compilation of a herbal medicine formulary for herbal substances in Malta and its usefulness amongst healthcare professionals.

CONTEXT: Today, the use of herbal medicine for primary healthcare has increased considerably. Since local pharmacists graduate with little knowledge on herbal medicine, the majority are ill-equipped to provide pharmaceutical advice. AIMS: To develop and evaluate a herbal medicine formulary to aid healthcare professionals (HCPs) in the prescribing, dispensing and counselling responsibilities. SETTINGS AND DESIGN: Community pharmacies. METHODS AND MATERIAL: Monographs on all herbal substances available locally were compiled into a formulary. The formulary was then distributed to all, 216, local pharmacies. Subsequently, a questionnaire was distributed to 55 pharmacists and 10 general practitioners (GPs). STATISTICAL ANALYSIS USED: Descriptive statistical analysis. RESULTS: A total of 177 herbal monographs have been compiled and 612 herbal products listed. Thirty HCPs participated in the questionnaire. The formulary was found to be useful by all participants with 19 claiming to use it frequently and 7 quite frequently. Participants (n = 30) agree that the information contained within the formulary was found to be useful (26), the formulary helped them learn which HMPs are present in the local market (29), the formulary is user friendly (27), information included is up-to-date and well referenced (29) and that there is the need for a formulary of this kind in Malta (28). CONCLUSIONS: The formulary was found to be a useful tool for HCPs leading to high quality, evidence-based prescribing together with enhanced monitoring and improved patient care.