Proteinaceous inhibitors of α-amylase and α-glucosidase from common bean (Phaseolus vulgaris L.): biochemical characterization and phylogenetic analysis of Sardinian cultivars

The common bean Phaseolus vulgaris is a good source of protein fiber and bioactive compounds. Among these proteinaceous α-amylase inhibitors (α-AIs) and α-glucosidase inhibitor could play an important role in weight loss and control of glycemic index above all in overweight and diabetic subjects. In this study 10 poorly studied Italian P. vulgaris cultivars were screened about their content in α-amylase/glucosidase inhibitors, antinutritional factor phytohemagglutinin (PHA), and about the expression of α-AI gene and their phylogenetic relationship. All the cultivars presented α-glucosidase inhibitor activity, while α-AI was missing in two of them. Only Nieddone cultivar (ACC177) had no hemagglutination activity and its α-AI was extracted, purified and characterized. The purified inhibitor has a tetrameric structure with a molecular weight of about 42 kDa, a temperature optimum of about 40°C and two pH optima (5 and 6.5). Thermal stability is remarkably important, since the inhibitory activity was maintained at 25% after 5 hours of incubation at 100°C. The purified inhibitor had the highest affinity toward insect α-amylase about 3-fold more susceptible than human salivary amylase and porcine pancreatic α-amylase. The kinetic characterization showed a mixed-type inhibition, suggesting a multiple site protein/protein interaction between enzyme (α-amylase) and inhibitor. Molecular studies showed that α-AI is expressed in all cultivars and a close similarity between the two cultivars Pisu Grogu and Fasolu α-AI and isoform α-AI-4 emerged from the comparison of the partially reconstructed primary structures. Moreover, from mechanistic models emerge the interaction network that connects the αAI with the α-amylase enzymecharacterized from two interaction hotspots (Asp38, and Tyr186), giving some insights for the analysis of α-AI primary structure from the different cultivars, particularly about structure-activity relationship. This study can be lay the basis for increase the knowledge about this class of proteins and developing commercial preparations from Italian common beans cultivars

Anticancer properties of bromelain: State-of-the-art and recent trends

Bromelain is a key enzyme found in pineapple (Ananas comosus (L.) Merr.); a proteolytic substance with multiple beneficial effects for human health such as anti-inflammatory, immunomodulatory, antioxidant and anticarcinogenic, traditionally used in many countries for its potential therapeutic value. The aim of this updated and comprehensive review focuses on the potential anticancer benefits of bromelain, analyzing the cytotoxic, apoptotic, necrotic, autophagic, immunomodulating, and anti-inflammatory effects in cancer cells and animal models. Detailed information about Bromelain and its anticancer effects at the cellular, molecular and signaling levels were collected from online databases such as PubMed/MedLine, TRIP database, GeenMedical, Scopus, Web of Science and Google Scholar. The results of the analyzed studies showed that Bromelain possesses corroborated pharmacological activities, such as anticancer, anti-edema, anti-inflammatory, anti-microbial, anticoagulant, anti-osteoarthritis, anti-trauma pain, anti-diarrhea, wound repair. Nonetheless, bromelain clinical studies are scarce and still more research is needed to validate the scientific value of this enzyme in human cancer diseases

Sisymbrium officinale, the Plant of Singers: A Review of Its Properties and Uses

Sisymbrium officinale (hedge mustard and formerly called Erysimum officinale) is a common plant in wild lands of Europe and Africa. It is also cultivated for its seeds and leaves to be used in salad or mustard. Sisymbrium officinale is useful not only in culinary preparations, but it also seems to possess interesting therapeutic properties, especially for throat diseases such as aphonia and hoarseness. For this reason, it is commonly called “herb of singers” (in Italian, “Erba dei cantanti”). Indeed a cup of Sisymbrium officinale infusion is frequently consumed by singers before artistic performance, even if its beneficial ability still needs to be scientifically demonstrated. Some preliminary data can be analyzed, but new efforts and resources should be devoted to study and investigate a plant with valuable therapeutic potential. This review summarizes the data available for Sisymbrium officinale

Plants as a Promising Reservoir of Tyrosinase Inhibitors

The process of melanogenesis, that takes place in the melanocytes of the epidermis, leads to hyperpigmentation. The biosynthetic pathway for production of melanin involves the enzyme tyrosinase that has been an attractive target for cosmaceutical research. Numerous synthetic, semisynthetic and natural, especially plant-based, inhibitors of tyrosinase have been reported in the literature. In plants, the secondary metabolites like flavonoids, chalcones, stilbenes, tannins, hydroquinone and kojic acid, etc... have been shown to possess the anti-tyrosinase activity. In the current review, we have covered the progress in this sphere that would be useful for not only further mechanistic investigations but also for the optimization of the structure of the metabolites for improved activity and selectivity. Thus the review presents a comprehensive report on tyrosinase inhibitors of plant origin reported in the extract form or as isolated compounds. Huge gap has been found between research and industry due to inconsistent pursual of the potent plant based extracts. There is a need to completely evaluate the extracts for structure optimization using molecular docking and evaluation of the safety inorder to benefit the industry with non toxic biological friendly products through invivo and exvivo optimization

The success in the short-distance communication for mating does not depend on chemical signals in the crustacean decapod Procambarus clarkii (Girard, 1852)

Pheromone-driven sex recognition has been widely documented in crayfish and a great deal of evidence supports the involvement of pheromones in their mating behaviour. This study investigates whether sexual interaction and mating success in the red swamp crayfish Procambarus clarkii are dependent on short-distance chemical communication between sexes, mediated by urine-borne pheromones. We compared the mating behaviour of intact animals that could release urine to chemically communicate in a natural way with that of urine-blocked animal pairs, for which chemical communication was precluded. Our results show that urine-borne pheromones are not critical for the reproductive success of P. clarkii, at least over the short-range distance (<1 m) considered in this study, during which the animals were confined in a restricted tank, facing one each other, and thus able to promptly mate. Under these experimental conditions, a lack of urine release neither precluded the occurrence, nor affected the duration of the different phases of mating behaviour. We conclude that short-distance chemical communication in P. clarkii is not a prerequisite for mating, and suggest that it could be affected by alternative sensory modalities, likely vision and/or acoustic signalling

Development of PVC Dispensers for Long-Lasting Release of Attractants for the Control of Invasive Crayfish Populations

Invasive alien species (IAS) are considered one of the major threats to biodiversity worldwide, thus requiring severe control strategies. Based on the promising results obtained in the field of insect pest management with polyvinyl chloride (PVC) for long-lasting release of attractants, the main aim of this study was to ascertain the efficacy of PVC/attractant dispensers also in the aquatic environment. Therefore, we developed PVC/food dispensers and evaluated their attractiveness, by means of behavioural bioassays on whole animals, over a 60-day period of continuous use towards the red swamp crayfish Procambarus clarkii, one of the top 100 worst IAS. The attractiveness in PVC of trehalose, leucine and taurocholic acid was also tested. Our results show that the PVC dispensers release the food and are strongly attractive for crayfish over a prolonged time, even if their effectiveness depends on the storage conditions: From 18 days when stored underwater at 23 °C up to 50 days when stored out of the water at -20 °C. Besides, trehalose, leucine and taurocholic acid in PVC resulted in reliable attractants for P. clarkii. The development of PVC dispensers for long-lasting release of attractants may help improve the efficiency of mass trapping strategies in the management and control of invasive crayfish

Biochemical and Phylogenetic Analysis of Italian <i>Phaseolus vulgaris</i> Cultivars as Sources of α-Amylase and α-Glucosidase Inhibitors

Phaseolus vulgaris α-amylase inhibitor (α-AI) is a protein that has recently gained commercial interest, as it inhibits mammalian α-amylase activity, reducing the absorption of dietary carbohydrates. Numerous studies have reported the efficacy of preparations based on this protein on the control of glycaemic peaks in type-2 diabetes patients and in overweight subjects. A positive influence on microbiota regulation has also been described. In this work, ten insufficiently studied Italian P. vulgaris cultivars were screened for α-amylase- and α-glucosidase-inhibiting activity, as well as for the absence of antinutritional compounds, such as phytohemagglutinin (PHA). All the cultivars presented α-glucosidase-inhibitor activity, while α-AI was missing in two of them. Only the Nieddone cultivar (ACC177) had no haemagglutination activity. In addition, the partial nucleotide sequence of the α-AI gene was identified with the degenerate hybrid oligonucleotide primer (CODEHOP) strategy to identify genetic variability, possibly linked to functional α-AI differences, expression of the α-AI gene, and phylogenetic relationships. Molecular studies showed that α-AI was expressed in all the cultivars, and a close similarity between the Pisu Grogu and Fasolu cultivars’ α-AI and α-AI-4 isoform emerged from the comparison of the partially reconstructed primary structures. Moreover, mechanistic models revealed the interaction network that connects α-AI with the α-amylase enzyme characterized by two interaction hotspots (Asp38 and Tyr186), providing some insights for the analysis of the α-AI primary structure from the different cultivars, particularly regarding the structure–activity relationship. This study can broaden the knowledge about this class of proteins, fuelling the valorisation of Italian agronomic biodiversity through the development of commercial preparations from legume cultivars

Small-scale crayfish distribution modelling of the invasive crayfish Procambarus clarkii in the Molentargius-Saline Regional Natural Park (Sardinia, Italy) in response to environmental variables

The crayfish, Procambarus clarkii, is among the worst invasive species capable of disperse overland, aiding the invasion of new environments. Because management options are limited once crayfish become established, there is a critical need to identify the suitable habitats for this invasive species. In this study, we forecast the potential distribution of crayfish based on environmental predictors in the Molentargius-Saline Regional Natural Park (Sardinia, Italy). Current known distribution was used to build correlative species distribution models (SDMs) and forecast distribution in the entire area. We considered, during the spring season, 5 quantitative environmental descriptors at the local scale (Depth, Water Temperature, Salinity, pH and Dissolved Oxygen). Single station data were used to generate synoptic distribution of environmental factors through the inverse distance weighted deterministic interpolation (IDW). The SDMs obtained through 2 different algorithms showed very good performance: GBM (General Boosted Models, ROC=0.847) and RF (Random Forest, ROC=0.996). For GBM the predicted distribution correlated most significantly with environmental variables (Salinity, Dissolved Oxygen and pH), while for RF all variables are homogeneously significant. Identified best models, performed by the Ensemble Modeling technique, showed that most of the areas outside the currently known distribution may offer potential habitats for P. clarkii. Greater probability to find the species presence were computed in correspondence of the Deflux Channel and ESF6B basin of the EcoSistema Filtro. However, for the Bellarosa Minore basin the forecast spread of the species is evident in all the tanks with large dense patches. Presented results deepen the knowledge about distribution patterns of P. clarkii in the Regional Natural Park. Moreover, the ongoing study gives a basic background for the future development of management strategies. This study was supported by the Fondazione di Sardegna for the project Impact of Invasive Alien Species on Sardinian ecosystems and by the Fondazione con il Sud for the project Nr. 2015-AMB-0065

The imbalance of serotonergic circuitry impairing the crop supercontractile muscle activity and the mitochondrial morphology of PD PINK1B9 Drosophila melanogaster are rescued by Mucuna pruriens

Despite its great potentiality, little attention has been paid to modelling gastrointestinal symptoms of Parkinson's disease (PD) in Drosophila melanogaster (Dm). Our previous studies on standardized Mucuna pruriens extract (Mpe) have shown usefulness in the Drosophila model of PD. In this communication, we provide new information on the effect of Mpe on basal and serotonin treated contractions in the crop (i.e., an important and essential part of the gut) in Drosophila PD mutant for PTEN-induced putative kinase 1 (PINK1B9) gene. The effect of Mpe on PINK1B9 supplied with standard diet to larvae and/or adults, were assayed on 10–15 days old flies. Conversely from what we observed in the wild type flies, recordings demonstrated that exogenous applications of serotonin on crop muscles of untreated PINK1B9 affect neither the frequency nor the amplitude of the crop contraction, while the same muscle parameters are enhanced following brain injections of serotonin, thus suggesting that PINK1B9 mutants may likely have an impairment in the serotonergic pathways. Also, the mitochondrial morphology in the crop muscles is strongly compromised, as demonstrated by the transmission electron microscopy analysis. The Mpe treatment rescued the crop muscle parameters and also the mitochondrial morphology when supplied to both larvae and adults. Overall, this study strengthens the relevance of using PINK1B9 Dm as a translational model to study the gastrointestinal symptoms in PD and also confirms the useful employment of M. pruriens for PD treatment