Summary• NADP‐glutamate dehydrogenase (NADP‐GDH) from Tuber borchii was purified and the correspo... more Summary• NADP‐glutamate dehydrogenase (NADP‐GDH) from Tuber borchii was purified and the corresponding gene was cloned in order to elucidate the physiological role of the enzyme in this ectomycorrhizal fungus.• NADP‐GDH was purified using an anion‐exchange column followed by affinity chromatography. The complete gene was cloned from a 30‐d‐old‐mycelium cDNA library and characterized.• T. borchii NADP‐GDH appears to be physically and kinetically similar to those from other fungi and the deduced amino acid sequence of the gdh gene showed a significant similarity to other fungal NADP‐dependent GDHs. Biochemical and Northern blotting analyses carried out with mycelia grown on different nitrogen sources clearly showed that the regulation of T. borchii NADP‐GDH in response to different nitrogen sources was markedly different from the responses of the NADP‐GDHs of other ascomycetes. Northern blotting analyses highlighted that the gdh gene was also expressed in the symbiotic phase.• The biochemical and molecular data suggest that the fungal NADP‐GDH contributes to the primary nitrogen metabolism in the ectomycorrhizal tissues.
During the different phases of mycorrhizal fungi life cycle, several morphological, genetic and m... more During the different phases of mycorrhizal fungi life cycle, several morphological, genetic and metabolic modifications are induced for both symbiotic partners. These changes are influenced by multiple environmental factors: light, gravity, oxygen, temperature, soil typology, nutrient availability, root exudates (that may content sugars) and the presence of particular bacterial populations in the mycorrhizosphere. Herein, in order to verify if nutritional supply leads to cell signaling events and morpho-functional changes in Tuber borchii pure culture, the expression level of genes involved in the apical growth, was investigated using the array technique, on mycelia grown on culture media containing different carbon sources. Membranes were prepared using ESTs involved in polarized growth, such as: S97, codifying the protein sec1, responsible for binding Syntaxin and involved in vesicle membrane fusion; S102, involved in apical growth, that codify a 1,3-beta-glucanosil transferase responsible for cell wall assembly of fungi; S32, directly involved in cell nuclear migration towards hyphae apical tips by cytoskeleton. The results obtained by morphological analyses highlighted an increased growth of T. borchii mycelium in the presence of glucose, while the hyphae are thinner and less branched in the presence of sucrose and maltose. These data were confirmed by gene expression analyses: the genes involved in apical grown and general metabolism are more expressed. Since glucose is also present in root plant exudates, it can act as a signal for the fungus to indicate the presence of the plant, and to induce the complex symbiotic process. These mechanisms can lead to morphological modifications, among which an increase branching, necessary for the fungus to get closer to the plant roots and establish a contact. This study represents a starting point to identify and characterise genes and proteins involved in the complex morphological, genetic and metabolic modifications that happens during Tuber life cycle
A third partner in the symbiosis between the fungus and plant root is represented by natural bact... more A third partner in the symbiosis between the fungus and plant root is represented by natural bacterial communities, which seem to play pivotal role in the complex biological processes of exchange involving nutrients and signaling from the soil hyphae, ectomycorrhizas, ascomata, and stromata. This review summarizes the recent evidence reported in literature showing that ascoma provides a habitat to complex microbial communities that are clearly differentiated from those of the soil and the ectomycorrhizosphere. Although the traditional plate isolation and the culture of microbes are indispensable for vegetative compatibility and/or functional assays, these techniques do not let a real in vivo picture the truffle ecosystem. Advent of the next-generation sequencing methods and recent advances in microarray technologies have increased culture-independent studies. Indeed, most microbiota remains uncultivable in laboratory conditions, and these novel technologies have greatly improved the understanding of microbial diversity and its functioning. This is particularly important for the ectomycorrhizal fungi of Tuber genus, since specific uncultivable-associated prokaryotes may play important roles in the biological system and ontogenetic cycle of these fungi. Success of truffle cultivation may be achieved with co-inoculum of specific bacteria with Tuber spp. upon establishment of commercial plantations; this is particularly relevant to the Tuber species with a high agronomic value.
3° Congresso Internazionale di Spoleto sul Tartufo, 2010
The first insight into T. borchii isoprenoid pathway, also known as mevalonic acids pathway (MVA)... more The first insight into T. borchii isoprenoid pathway, also known as mevalonic acids pathway (MVA) during ascoma maturation were here discussed . Three genes coding for the most important enzymes of the isoprenoid biosynthesis were characterised, and the activity of the major regulatory enzyme of the MVA was determined in particular during the ascomata maturation. Furthermore several isoprenoids were identified among the volatile compounds produced by ripe T. borchii ascomat
Background: MoviS: ‘Movement and Health Beyond Care’ is a 3-year randomized controlled trial aimi... more Background: MoviS: ‘Movement and Health Beyond Care’ is a 3-year randomized controlled trial aiming to educate cancer patients after adjuvant therapy on the benefits of personalized physical activity (PA) and a proper nutritional plan Methods: In this project, breast cancer (BC) patients who consent to participate will be randomized to Interventional Arm (A), consisting of 3-months of Movis Training, or Control Arm (B), consisting of standard care with no supervised PA The Movis Training consists of 3-months of aerobic training (2 d/week of supervised training and 1 d/week of unsupervised exercise) with an increase in exercise intensity (40-70% Heart Rate Reserve) and duration (20-60 min) Ad interim analyses every 3-months up to 1-year will be included The first cohort of eligible BC survivors were enrolled in January 2020 and carried out the Movis Training even during the COVID-19 pandemic The primary outcome is improvement of Quality of Life (QoL) assessed by European Organization for Research and Treatment of Cancer QoL (EORTC QLQ-C30) The secondary outcomes are improvement of health-related QoL parameters such as: PA level (International Physical Activity Questionnaire;SenseWear Armband), fitness (VO2max), flexibility, strength, psychological well-being (Psychological distress Inventory;Profile of Mood States and diet habit (DianaWeb, MEDIET modified and recall 24h);Anthropometric measurement, Body mass (kg);BMI (kg/m2);body composition Results: The expected improvement (mean ± SD) of the QoL in Arm A at 3-months is 15 1 ± 17 7, while in Arm B is 6 1 ± 17 1 (Cohen d effect size=0 51, medium effect) Using a t test for independent samples, with 0 05 alpha and 0 80 1-beta will require 60 subjects per group Considering an expected drop-out of 30%, a total of 172 patients will be recruited Conclusions: The targeted exercise oncology through multidisciplinary team would like to provide a coordinated program of cancer care to improve health care quality, improve prognosis, increase survival times and QoL and reduce the risk of BC recurrence Legal entity responsible for the study: Elena Barbieri Funding: Has not received any funding Disclosure: All authors have declared no conflicts of interest
BackgroundBreast cancer (BC) is the second-leading cause of cancer-related death worldwide. This ... more BackgroundBreast cancer (BC) is the second-leading cause of cancer-related death worldwide. This study aimed to investigate the effects of a 12-week home-based lifestyle intervention (based on nutrition and exercise) on gut microbial composition in twenty BC survivors of the MoviS clinical trial (protocol: NCT 04818359).MethodsGut microbiota analysis through 16S rRNA gene sequencing, anthropometrics, Mediterranean Diet (MD) adherence, and cardiometabolic parameters were evaluated before (Pre) and after (Post) the lifestyle intervention (LI).ResultsBeneficial effects of the LI were observed on MD adherence, and cardiometabolic parameters (pre vs post). A robust reduction of Proteobacteria was observed after LI, which is able to reshape the gut microbiota by modulating microorganisms capable of decreasing inflammation and others involved in improving the lipid and glycemic assets of the host. A significant negative correlation between fasting glucose and Clostridia_vadinBB60 (r = -0.6...
Summary• NADP‐glutamate dehydrogenase (NADP‐GDH) from Tuber borchii was purified and the correspo... more Summary• NADP‐glutamate dehydrogenase (NADP‐GDH) from Tuber borchii was purified and the corresponding gene was cloned in order to elucidate the physiological role of the enzyme in this ectomycorrhizal fungus.• NADP‐GDH was purified using an anion‐exchange column followed by affinity chromatography. The complete gene was cloned from a 30‐d‐old‐mycelium cDNA library and characterized.• T. borchii NADP‐GDH appears to be physically and kinetically similar to those from other fungi and the deduced amino acid sequence of the gdh gene showed a significant similarity to other fungal NADP‐dependent GDHs. Biochemical and Northern blotting analyses carried out with mycelia grown on different nitrogen sources clearly showed that the regulation of T. borchii NADP‐GDH in response to different nitrogen sources was markedly different from the responses of the NADP‐GDHs of other ascomycetes. Northern blotting analyses highlighted that the gdh gene was also expressed in the symbiotic phase.• The biochemical and molecular data suggest that the fungal NADP‐GDH contributes to the primary nitrogen metabolism in the ectomycorrhizal tissues.
During the different phases of mycorrhizal fungi life cycle, several morphological, genetic and m... more During the different phases of mycorrhizal fungi life cycle, several morphological, genetic and metabolic modifications are induced for both symbiotic partners. These changes are influenced by multiple environmental factors: light, gravity, oxygen, temperature, soil typology, nutrient availability, root exudates (that may content sugars) and the presence of particular bacterial populations in the mycorrhizosphere. Herein, in order to verify if nutritional supply leads to cell signaling events and morpho-functional changes in Tuber borchii pure culture, the expression level of genes involved in the apical growth, was investigated using the array technique, on mycelia grown on culture media containing different carbon sources. Membranes were prepared using ESTs involved in polarized growth, such as: S97, codifying the protein sec1, responsible for binding Syntaxin and involved in vesicle membrane fusion; S102, involved in apical growth, that codify a 1,3-beta-glucanosil transferase responsible for cell wall assembly of fungi; S32, directly involved in cell nuclear migration towards hyphae apical tips by cytoskeleton. The results obtained by morphological analyses highlighted an increased growth of T. borchii mycelium in the presence of glucose, while the hyphae are thinner and less branched in the presence of sucrose and maltose. These data were confirmed by gene expression analyses: the genes involved in apical grown and general metabolism are more expressed. Since glucose is also present in root plant exudates, it can act as a signal for the fungus to indicate the presence of the plant, and to induce the complex symbiotic process. These mechanisms can lead to morphological modifications, among which an increase branching, necessary for the fungus to get closer to the plant roots and establish a contact. This study represents a starting point to identify and characterise genes and proteins involved in the complex morphological, genetic and metabolic modifications that happens during Tuber life cycle
A third partner in the symbiosis between the fungus and plant root is represented by natural bact... more A third partner in the symbiosis between the fungus and plant root is represented by natural bacterial communities, which seem to play pivotal role in the complex biological processes of exchange involving nutrients and signaling from the soil hyphae, ectomycorrhizas, ascomata, and stromata. This review summarizes the recent evidence reported in literature showing that ascoma provides a habitat to complex microbial communities that are clearly differentiated from those of the soil and the ectomycorrhizosphere. Although the traditional plate isolation and the culture of microbes are indispensable for vegetative compatibility and/or functional assays, these techniques do not let a real in vivo picture the truffle ecosystem. Advent of the next-generation sequencing methods and recent advances in microarray technologies have increased culture-independent studies. Indeed, most microbiota remains uncultivable in laboratory conditions, and these novel technologies have greatly improved the understanding of microbial diversity and its functioning. This is particularly important for the ectomycorrhizal fungi of Tuber genus, since specific uncultivable-associated prokaryotes may play important roles in the biological system and ontogenetic cycle of these fungi. Success of truffle cultivation may be achieved with co-inoculum of specific bacteria with Tuber spp. upon establishment of commercial plantations; this is particularly relevant to the Tuber species with a high agronomic value.
3° Congresso Internazionale di Spoleto sul Tartufo, 2010
The first insight into T. borchii isoprenoid pathway, also known as mevalonic acids pathway (MVA)... more The first insight into T. borchii isoprenoid pathway, also known as mevalonic acids pathway (MVA) during ascoma maturation were here discussed . Three genes coding for the most important enzymes of the isoprenoid biosynthesis were characterised, and the activity of the major regulatory enzyme of the MVA was determined in particular during the ascomata maturation. Furthermore several isoprenoids were identified among the volatile compounds produced by ripe T. borchii ascomat
Background: MoviS: ‘Movement and Health Beyond Care’ is a 3-year randomized controlled trial aimi... more Background: MoviS: ‘Movement and Health Beyond Care’ is a 3-year randomized controlled trial aiming to educate cancer patients after adjuvant therapy on the benefits of personalized physical activity (PA) and a proper nutritional plan Methods: In this project, breast cancer (BC) patients who consent to participate will be randomized to Interventional Arm (A), consisting of 3-months of Movis Training, or Control Arm (B), consisting of standard care with no supervised PA The Movis Training consists of 3-months of aerobic training (2 d/week of supervised training and 1 d/week of unsupervised exercise) with an increase in exercise intensity (40-70% Heart Rate Reserve) and duration (20-60 min) Ad interim analyses every 3-months up to 1-year will be included The first cohort of eligible BC survivors were enrolled in January 2020 and carried out the Movis Training even during the COVID-19 pandemic The primary outcome is improvement of Quality of Life (QoL) assessed by European Organization for Research and Treatment of Cancer QoL (EORTC QLQ-C30) The secondary outcomes are improvement of health-related QoL parameters such as: PA level (International Physical Activity Questionnaire;SenseWear Armband), fitness (VO2max), flexibility, strength, psychological well-being (Psychological distress Inventory;Profile of Mood States and diet habit (DianaWeb, MEDIET modified and recall 24h);Anthropometric measurement, Body mass (kg);BMI (kg/m2);body composition Results: The expected improvement (mean ± SD) of the QoL in Arm A at 3-months is 15 1 ± 17 7, while in Arm B is 6 1 ± 17 1 (Cohen d effect size=0 51, medium effect) Using a t test for independent samples, with 0 05 alpha and 0 80 1-beta will require 60 subjects per group Considering an expected drop-out of 30%, a total of 172 patients will be recruited Conclusions: The targeted exercise oncology through multidisciplinary team would like to provide a coordinated program of cancer care to improve health care quality, improve prognosis, increase survival times and QoL and reduce the risk of BC recurrence Legal entity responsible for the study: Elena Barbieri Funding: Has not received any funding Disclosure: All authors have declared no conflicts of interest
BackgroundBreast cancer (BC) is the second-leading cause of cancer-related death worldwide. This ... more BackgroundBreast cancer (BC) is the second-leading cause of cancer-related death worldwide. This study aimed to investigate the effects of a 12-week home-based lifestyle intervention (based on nutrition and exercise) on gut microbial composition in twenty BC survivors of the MoviS clinical trial (protocol: NCT 04818359).MethodsGut microbiota analysis through 16S rRNA gene sequencing, anthropometrics, Mediterranean Diet (MD) adherence, and cardiometabolic parameters were evaluated before (Pre) and after (Post) the lifestyle intervention (LI).ResultsBeneficial effects of the LI were observed on MD adherence, and cardiometabolic parameters (pre vs post). A robust reduction of Proteobacteria was observed after LI, which is able to reshape the gut microbiota by modulating microorganisms capable of decreasing inflammation and others involved in improving the lipid and glycemic assets of the host. A significant negative correlation between fasting glucose and Clostridia_vadinBB60 (r = -0.6...
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