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Fluorescence-Based Multiplex Western Blot to Simultaneously Detect the Insulin-Like Growth Factor-1 (IGF-1) Isoforms
No full text: Insulin-like growth factor-1 (IGF-1) is critical for tissue growth and development. The IGF-1 gene contains six exons and due to alternative splicing three different isoforms might be produced: the IGF-1Ea, Eb, and Ec prohormones (proIGF-1s). These proIGF-1s share the same IGF-1 mature sequence, which is responsible for the IGF-1 receptor binding but differ in their carboxy-terminal extensions called Ea-, Eb-, and Ec-peptides. Several lines of evidence indicate that E-peptides control the intracellular proIGF-1s localization and maturation. Here, we present a multiplex Western blotting system able to simultaneously discriminate and quantify mature IGF-1, proIGF-1s and E-peptides within the same sample. HEK293 cells were transiently transfected with plasmids containing the IGF-1Ea, IGF-1Eb, or IGF-1Ec isoform or an empty vector. Two different primary antibodies, which recognize the mature sequence or the common region of E-peptides, were used to detect IGF-1 isoforms, which were subsequently distinguished with secondary antibodies conjugated to different fluorophores. Our results demonstrate the feasibility of simultaneously detecting different IGF-1 isoforms using two primary antibodies directed against different epitopes of proIGF-1s, combined with fluorescence-conjugated secondary antibodies. Furthermore, this dual-epitope strategy increases the specificity of protein detection, making it a valuable tool for studying the diverse roles of IGF-1 isoforms in biological processes
The isoprenoid pathway in the ectomycorrhizal fungus Tuber borchii Vittad.: cloning and characterisation of the tbhmgr, tbfpps and tbsqs genes.
No full textUse of Streptococcus salivarius K12 in supporting the mucosal immune function of active young subjects: A randomised double-blind study
Get PDFIntroduction: Upper respiratory tract infections (URTI) are the most common illnesses affecting athletes, causing absences from training and competition. Salivary immunoglobulin A (sIgA) is the main immune factor in saliva, and a consistent association between low concentrations of sIgA and an increased incidence of URTIs has been reported. The oral probiotic Streptococcus salivarius K12 has been suggested to have the potential to improve oral diseases and mucosal barrier function. However, the effects of this probiotic on active young subjects performing a high-intensity training (HIT) program have not been investigated. Methods: Active young students were randomised into a treated group (S. salivarius K12) and a control (placebo) group and asked to take the product daily for 30 days. After this period, participants performed a graded exercise test and five HIT sessions, all within 3 days. They were also asked to complete the Wisconsin Upper Respiratory Symptom Survey daily to monitor URTI's presence. Before and after the 30 days, and at 0h, 24h and 72h after the last training session, saliva samples were collected to quantify sIgA level, secretion rate, and flow. The effect of S. salivarius K12 intake on these parameters was tested using an ANOVA for repeated measures. Results: Twenty (M = 14, F = 6) young subjects (23.5 ± 2.3 years old) participated in the study. The total accumulated training load (sRPE) in the supplementation period was similar in the two groups (treated: 4345 ± 3441 AU; control: 4969 ± 4165 AU; p > 0.05). Considering both sIgA level and secretion rate, significant time (F(4,15) = 3.38; p = 0.037; F(4,15) = 6.00; p = 0.004) and time×group interactions (F(4,15) = 2.49; p = 0.049; F(4,15) = 5.01; p = 0.009) were reported, with the treated group showing higher sIgA levels at 72h post-exercise and increased secretion rate both at 0h and 72h. The number of URTI episodes was similar in the treated and control groups (χ2 = 1.83; p > 0.05). Conclusion: This study demonstrates that relatively short-term S. salivarius K12 supplementation increased sIgA level and secretion in healthy subjects performing a demanding exercise-training programme composed of HIT sessions
Sarcopenia ed esercizio fisico
No full textIn questo articolo viene descritto il ruolo dell’esercizio fisico nel migliorare la funzionalità e la dinamicità mitocondriale e nel favorire una buona qualità dell’invecchiamento muscolare; si tratta di un’area della ricerca bioenergetica che pone i mitocondri al centro delle teorie sull’invecchiamento
Mutual Interactions among Exercise, Sport Supplements and Microbiota
Get PDFThe adult gut microbiota contains trillions of microorganisms of thousands of differentspecies. Only one third of gut microbiota are common to most people; the rest are specific andcontribute to enhancing genetic variation. Gut microorganisms significantly affect host nutrition,metabolic function, immune system, and redox levels, and may be modulated by several environmentalconditions, including physical activity and exercise. Microbiota also act like an endocrine organ andis sensitive to the homeostatic and physiological changes associated with training; in turn, exercisehas been demonstrated to increase microbiota diversity, consequently improving the metabolic profileand immunological responses. On the other side, adaptation to exercise might be influenced bythe individual gut microbiota that regulates the energetic balance and participates to the control ofinflammatory, redox, and hydration status. Intense endurance exercise causes physiological andbiochemical demands, and requires adequate measures to counteract oxidative stress, intestinalpermeability, electrolyte imbalance, glycogen depletion, frequent upper respiratory tract infections,systemic inflammation and immune responses. Microbiota could be an important tool to improveoverall general health, performance, and energy availability while controlling inflammation andredox levels in endurance athletes. The relationship among gut microbiota, general health, trainingadaptation and performance, along with a focus on sport supplements which are known to exertsome influence on the microbiota, will be discussed
Effect of androgen exposure on inflammatory response of male and female vascular endothelial cells.
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Effects of sex hormones on inflammatory response in male and female vascular endothelial cells
Get PDFPURPOSE:
Gender-related differences in sex hormones might have a key role in the development of atherosclerosis though direct vascular effects of sex hormones are not yet well understood. Thus, the main purpose of this study was to compare the effects of sex hormones on inflammatory response in Human Umbilical Vein Endothelial Cells (HUVECs) obtained from both male and female donors.
METHODS:
We analyzed the expression of receptors and enzymes relevant to the action of androgens (AR, 5α-reductase 1 and 5α-reductase 2) and estrogens (ERα, ERβ, and aromatase) in male and female HUVECs. Furthermore, we analyzed the effect of testosterone (T), 17β-estradiol (E2), dihydrotestosterone (DHT), and several androgenic-anabolic steroids (AAS) on VCAM-1, ICAM-1, and E-selectin gene expression and on adhesion of U937 cells to TNF-α-stimulated male and female HUVECs.
RESULTS:
Our results reveal that in HUVECs, regardless of gender, the components involved in the androgen action pathway are predominant as compared to those of estrogen action pathway. In both HUVEC genders, the inflammatory effect of TNF-α was amplified by co-administration of T or DHT and several AAS frequently used in doping, while E2 had no effect.
CONCLUSIONS:
This is the first study analyzing, under identical culture conditions, the key components of sex hormone response in male and female HUVECs and the possible role of sex hormones in regulating the endothelial inflammatory response. The data obtained in our experimental system showed a pro-inflammatory effect of androgens, while conclusively excluding any protective effect for all the tested hormones
Effect of supervised aerobic and strength training in patients with type 2 diabetes mellitus
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