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A novel methodological approach to simultaneously extract high-quality total RNA and proteins from cortical and trabecular bone.
Faraldi, M, Mangiavini, L, Conte, C, Banfi, G, Napoli, N, Lombardi, G
Open biology. 2022;(5):210387
Abstract
Molecular differences between cortical and trabecular bone, of relevance to understanding the pathophysiological basis of bone diseases, can be determined only through effective isolation methods for RNA and proteins. Here we present a TRIzol-based method, which combines bone pulverization and homogenization to extract simultaneously total RNA and proteins from human cortical and trabecular bone from the same carrot. RNA integrity and purity were determined as the 260/280 nm and 260/230 nm absorbance ratios and the 28S/18S rRNA ratio. Protein integrity and quality were evaluated by Coomassie blue staining. Reverse transcription quantitative polymerase chain reaction and immunoblotting for bone-specific genes and proteins were performed to verify the suitability of the isolated material in downstream applications. The 260/280 nm and 260/230 nm absorbance ratios were, on average, less than or equal to 1.8. Bands on agarose gel were consistent with intact RNA, with mean 28S/18S ratios of 1.68 ± 0.35 and 1.88 ± 0.10 for cortical and trabecular bone, respectively. Band patterns after Coomassie blue staining confirmed protein integrity. Successful gene and protein expression analysis, with relevant differences between the two compartments, highlighted the suitability of the material in downstream applications. The method presented here is appropriate and effective for the study of human bone.
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Vitamin D in the Covid-19 era: a review with recommendations from a G.I.O.S.E.G. expert panel.
Ulivieri, FM, Banfi, G, Camozzi, V, Colao, A, Formenti, AM, Frara, S, Lombardi, G, Napoli, N, Giustina, A
Endocrine. 2021;(3):597-603
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Abstract
Vitamin D (VITD) is a key hormone for bone health and has relevant extra-skeletal effects that might play a role in the prevention and treatment of COronaVIrus Disease 19 (COVID-19). Literature regarding this scenario is voluminous but controversial. Glucocorticoid Induced Osteoporosis Skeletal Endocrinology Group (G.I.O.S.E.G) has been present in the scientific debate about the use of VITD and has regularly interfaced national regulatory agencies to ensure appropriateness of its employment. Given the current pandemic and the questions on COVID-19 and VITD, G.I.O.S.E.G. appointed an expert panel to advise how to consider this issue best. The results of these deliberations are reported in the current recommendation paper.
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Repetitive deep TMS for the reduction of body weight: Bimodal effect on the functional brain connectivity in "diabesity".
Devoto, F, Ferrulli, A, Zapparoli, L, Massarini, S, Banfi, G, Paulesu, E, Luzi, L
Nutrition, metabolism, and cardiovascular diseases : NMCD. 2021;(6):1860-1870
Abstract
BACKGROUND AND AIMS Deep repetitive Transcranial Magnetic Stimulation (deep rTMS) over the bilateral insula and prefrontal cortex (PFC) can promote weight-loss in obesity, preventing cardiometabolic complications as Type 2 Diabetes (T2D). To investigate the changes in the functional brain integration after dTMS, we conducted a resting-state functional connectivity (rsFC) study in obesity. METHODS AND RESULTS This preliminary study was designed as a randomized, double-blind, sham-controlled study: 9 participants were treated with high-frequency stimulation (realTMS group), 8 were sham-treated (shamTMS group). Out of the 17 enrolled patients, 6 were affected by T2D. Resting-state fMRI scans were acquired at baseline (T0) and after the 5-week intervention (T1). Body weight was measured at three time points [T0, T1, 1-month follow-up visit (FU1)]. A mixed-model analysis showed a significant group-by-time interaction for body weight (p = .04), with a significant decrease (p < .001) in the realTMS group. The rsFC data revealed a significant increase of degree centrality for the realTMS group in the medial orbitofrontal cortex (mOFC) and a significant decrease in the occipital pole. CONCLUSION An increase of whole-brain functional connections of the mOFC, together with the decrease of whole-brain functional connections with the occipital pole, may reflect a brain mechanism behind weight-loss through a diminished reactivity to bottom-up visual-sensory processes in favor of increased reliance on top-down decision-making processes. TRIAL REGISTRATION NUMBER ClinicalTrials.gov NCT03009695.
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Perceptual and Biochemical Responses in Relation to Different Match-Day +2 Training Interventions in Soccer Players.
Trecroci, A, Perri, E, Lombardi, G, Banfi, G, Del Vescovo, R, Rosa, EM, Alberti, G, Iaia, FM
Frontiers in physiology. 2021;:685804
Abstract
The aim of this study was to examine the impact of two different post-match training interventions on the subsequent recovery of perceptual and biochemical parameters after the game. In a crossover design, eight sub-elite players underwent a soccer-specific training (SST) and an active recovery (AR) regimen on the second day after a match (+48 h). Muscle soreness as well as muscle damage (creatine kinase, CK), inflammatory (C-reactive protein and interleukin 6), immunological (e.g., lymphocytes, neutrophils, and monocytes), and endocrine (cortisol) markers were obtained at baseline (-72 h), immediately after (0 h), and 72 h post-match (+72 h). AR promoted a higher restoration of muscle soreness values (P = 0.004, η2 p = 0.49) together with a better restoration of CK within 72 h post-match compared with SST (P = 0.04, η2 p = 0.36). Conversely, no significant (P > 0.05, η2 p < 0.91) differences were observed in the recovery timeframe of inflammatory, immunological, and endocrine responses between SST and AR. Overall, AR elicited a quicker muscle soreness and CK restoration compared to SST intervention at 72 h post-match. Such information provides novel evidence-based findings on the appropriateness of different recovery strategies and may aid to improve the practitioners' decision-making process when two consecutive games are played within 3 days.
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Hypocalcemia in COVID-19 is associated with low vitamin D levels and impaired compensatory PTH response.
di Filippo, L, Allora, A, Locatelli, M, Rovere Querini, P, Frara, S, Banfi, G, Giustina, A
Endocrine. 2021;(2):219-225
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Abstract
BACKGROUND Hypocalcemia has been identified as a major distinctive feature of COVID-19, predicting poor clinical outcomes. Among the mechanisms underlying this biochemical finding, high prevalence of vitamin D (VD) deficiency in COVID-19 patients reported so far in several studies was advocated. However, robust data in favor of this hypothesis are still lacking. Therefore, aim of our study was to investigate the role of hypovitaminosis D and parathyroid hormone (PTH) levels in the development of hypocalcemia in COVID-19 patients. METHODS Patients admitted to IRCCS Ospedale San Raffaele for COVID-19 were enrolled in this study, excluding those with comorbidities and therapies influencing calcium and VD metabolism. Serum levels of total calcium (tCa), ionized calcium (Ca2+), 25-OH-VD, and PTH were evaluated at admission. We defined VD deficiency as VD below 20 ng/mL, hypocalcemia as tCa below 2.2 mmol/L or as Ca2+ below 1.18 mmol/L, and hyperparathyroidism as PTH above 65 pg/mL. RESULTS A total of 78 patients were included in the study. Median tCa and Ca2+ levels were 2.15 and 1.15 mmol/L, respectively. Total and ionized hypocalcemia were observed in 53 (67.9%) and 55 (70.5%) patients, respectively. VD deficiency was found in 67.9% of patients, but secondary hyperparathyroidism was detected in 20.5% of them, only. tCa levels were significantly lower in patients with VD deficiency and regression analyses showed a positive correlation between VD and tCa. CONCLUSIONS In conclusion, we confirmed a high prevalence of hypocalcemia in COVID-19 patients and we showed for the first time that it occurred largely in the context of marked hypovitaminosis D not adequately compensated by secondary hyperparathyroidism.
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The Central Role of Iron in Human Nutrition: From Folk to Contemporary Medicine.
Briguglio, M, Hrelia, S, Malaguti, M, Lombardi, G, Riso, P, Porrini, M, Perazzo, P, Banfi, G
Nutrients. 2020;(6)
Abstract
Iron is a fundamental element in human history, from the dawn of civilization to contemporary days. The ancients used the metal to shape tools, to forge weapons, and even as a dietary supplement. This last indication has been handed down until today, when martial therapy is considered fundamental to correct deficiency states of anemia. The improvement of the martial status is mainly targeted with dietary supplements that often couple diverse co-factors, but other methods are available, such as parenteral preparations, dietary interventions, or real-world approaches. The oral absorption of this metal occurs in the duodenum and is highly dependent upon its oxidation state, with many absorption influencers possibly interfering with the intestinal uptake. Bone marrow and spleen represent the initial and ultimate step of iron metabolism, respectively, and the most part of body iron circulates bound to specific proteins and mainly serves to synthesize hemoglobin for new red blood cells. Whatever the martial status is, today's knowledge about iron biochemistry allows us to embrace exceedingly personalized interventions, which however owe their success to the mythical and historical events that always accompanied this metal.
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Home-Based Resistance Training for Older Subjects during the COVID-19 Outbreak in Italy: Preliminary Results of a Six-Months RCT.
Vitale, JA, Bonato, M, Borghi, S, Messina, C, Albano, D, Corbetta, S, Sconfienza, LM, Banfi, G
International journal of environmental research and public health. 2020;(24)
Abstract
BACKGROUND The aim of this study was to evaluate the effect of a six-month home-based resistance-training program on muscle health and physical performance in healthy older subjects during the unique condition of home confinement caused by the COVID-19 pandemic. METHODS This was a randomized-controlled study that enrolled older participants that were allocated to either an experimental group performing the six-months exercise prescription (EXE) or a control group (CON). At the beginning (PRE), and after 6 months (POST), participants were assessed for muscle strength, balance, gait assessment and body composition by dual energy X-ray absorptiometry and magnetic resonance imaging. Normality distribution of data was checked with the D'Agostino and Pearson test and changes between PRE and POST were assessed by paired Student's t-test while percentage and absolute changes between groups at POST were tested by unpaired t-test. RESULTS Nine participants were included for the final analysis: EXE, n = 5 (age: 66 ± 4; BMI: 27.5 ± 3.7) and CON, n = 4 (age: 71 ± 9; BMI: 24.2 ± 4.1). Significant PRE-to-POST changes were observed in the EXE group only in the chair-stand test (+19.8%, p = 0.048 and ES:1.0, moderate) and in total fat mass (+5.0%, p = 0.035 and ES:1.4, large) with no between-group differences. Moreover, EXE had significantly higher absolute thigh CSA values than CON at POST (14.138 ± 2977 vs. 9039 ± 1015, p = 0.0178, ES = 1.7). No other within- and between-group differences were detected. CONCLUSIONS The home-based resistance-training program during the lockdown period, caused by the COVID-19 outbreak, determined only within-group improvement in lower limb muscle strength but not in muscle mass and composition in older subjects. Home confinement may partially explain the increase in total body fat due to a reduced daily PA regime and altered diet pattern.
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Gastrointestinal In Vitro Digests of Infant Biscuits Formulated with Bovine Milk Proteins Positively Affect In Vitro Differentiation of Human Osteoblast-Like Cells.
Bottani, M, Cattaneo, S, Pica, V, Stuknytė, M, Gomarasca, M, Lombardi, G, Banfi, G, Noni, I, Ferraretto, A
Foods (Basel, Switzerland). 2020;(10)
Abstract
Infant biscuits (IBs) are part of complementary feeding from weaning up to the age of five years. They normally contain bovine milk proteins, which can influence bone development. This potential effect was investigated using experimental baked IBs, which were prepared from doughs containing different type of dairy proteins: milk protein concentrate (IB1), whey protein isolate (IB2), and skimmed milk powder (IB3). Dairy protein-free (IB0) and gluten-free (IB4) biscuits were also formulated. The in vitro gastrointestinal digests of IBs (IBDs) were tested on a co-culture of Caco-2/HT-29 70/30 cells as an in vitro model of human small intestine. None of the IBDs influenced cell viability and monolayer integrity, while IBD0 and IBD4 increased Peptide-YY production. The basolateral contents of Transwell plates seeded with Caco-2/HT-29 70/30 co-culture, mimicking metabolized IBDs (MIBDs), were tested on Saos-2 cells, an in vitro model of human osteoblast-like cells. After incubation, MIBD0, lacking dairy proteins, decreased the cell viability, while MIBD2, containing whey protein isolate, increased both the viability and the number of cells. MIBD2 and MIBD4, the latter containing both casein and whey proteins, increased alkaline phosphatase activity, a bone differentiation marker. These results highlight that IBs containing dairy proteins positively affect bone development.
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The Malnutritional Status of the Host as a Virulence Factor for New Coronavirus SARS-CoV-2.
Briguglio, M, Pregliasco, FE, Lombardi, G, Perazzo, P, Banfi, G
Frontiers in medicine. 2020;7:146
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Plain language summary
This opinion article explores the role of an individual’s nutrition status when subjected to infection by viruses, in particular Covid-19. Distinction is made between the susceptibility to infection in the first instance and the ability to persist in fighting infection once it is established. For Covid-19, it is argued that a healthier nutritional status, in particular Vitamins A, B, C, D and E, iron selenium and zinc, will lower susceptibility to infection, lower the severity of the virus and therefore reduce the length of time an individual has to find reserves to fight the virus. More severe cases of Covid-19 infection also often include gastro-intestinal symptoms which further exacerbate nutritional status with lowered appetite. The authors conclude that malnourished individuals may be more susceptible to Covid-19 infection and that nutritional support is vital in severe cases. The article includes a useful diagram of both hyponutrition and hypernutrition and possible impacts of Covid-19.
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Oral Supplementation with Sucrosomial Ferric Pyrophosphate Plus L-Ascorbic Acid to Ameliorate the Martial Status: A Randomized Controlled Trial.
Briguglio, M, Hrelia, S, Malaguti, M, De Vecchi, E, Lombardi, G, Banfi, G, Riso, P, Porrini, M, Romagnoli, S, Pino, F, et al
Nutrients. 2020;(2)
Abstract
Altered martial indices before orthopedic surgery are associated with higher rates of complications and greatly affect the patient's functional ability. Oral supplements can optimize the preoperative martial status, with clinical efficacy and the patient's tolerability being highly dependent on the pharmaceutical formula. Patients undergoing elective hip/knee arthroplasty were randomized to be supplemented with a 30-day oral therapy of sucrosomial ferric pyrophosphate plus L-ascorbic acid. The tolerability was 2.7% among treated patients. Adjustments for confounding factors, such as iron absorption influencers, showed a relevant response limited to older patients (≥ 65 years old), whose uncharacterized Hb loss was averted upon treatment with iron formula. Older patients with no support lost -2.8 ± 5.1%, while the intervention group gained +0.7 ± 4.6% of circulating hemoglobin from baseline (p = 0.019). Gastrointestinal diseases, medications, and possible dietary factors could affect the efficacy of iron supplements. Future opportunities may consider to couple ferric pyrophosphate with other nutrients, to pay attention in avoiding absorption disruptors, or to implement interventions to obtain an earlier martial status optimization at the population level.