Anastassios Philippou

IGF-1 Expression in Infarcted Myocardium and MGF E Peptide Actions in Rat Cardiomyocytes in Vitro

Insulinlike growth factor-1 (IGF-1) expression is implicated in myocardial pathophysiology, and two IGF-1 mRNA splice variants have been detected in rodents, IGF-1Ea and mechano-growth factor (MGF). We investigated the expression pattern of IGF-1 gene transcripts in rat myocardium from 1 h up to 8 wks after myocardial infarction induced by left anterior descending coronary artery ligation. In addition, we characterized IGF-1 and MGF E peptide action and their respective signaling in H9C2 myocardial-like cells in vitro. IGF-1Ea and MGF expression were significantly increased, both at transcriptional and translational levels, during the late postinfarction period (4 and 8 wks) in infarcted rat myocardium. Measurements of serum IGF-1 levels in infarcted rats were initially decreased (24 h up to 1 wk) but remained unaltered throughout the late experimental phase (4 to 8 wks) compared with sham-operated rats. Furthermore, specific anti-IGF-1R neutralizing antibody failed to block the synthetic MGF E peptide action, whereas it completely blocked IGF-1 action on the proliferation of H9C2 cells. Moreover, this synthetic MGF E peptide did not activate Akt phosphorylation, whereas it activated ERK1/2 in H9C2 rat myocardial cells. These data support the role of IGF-1 expression in the myocardial repair process and suggest that synthetic MGF E peptide actions may be mediated via an IGF-1R independent pathway in rat myocardial cells, as suggested by our in vitro experiments.

Short-term high-intensity interval exercise training attenuates oxidative stress responses and improves antioxidant status in healthy humans.

This study investigated the changes in oxidative stress biomarkers and antioxidant status indices caused by a 3-week high-intensity interval training (HIT) regimen. Eight physically active males performed three HIT sessions/week over 3 weeks. Each session included four to six 30-s bouts of high-intensity cycling separated by 4 min of recovery. Before training, acute exercise elevated protein carbonyls (PC), thiobarbituric acid reactive substances (TBARS), glutathione peroxidase (GPX) activity, total antioxidant capacity (TAC) and creatine kinase (CK), which peaked 24h post-exercise (252 ± 30%, 135 ± 17%, 10 ± 2%, 85 ± 14% and 36 ± 13%, above baseline, respectively; p<0.01), while catalase activity (CAT) peaked 30 min post-exercise (56 ± 18% above baseline; p<0.01). Training attenuated the exercise-induced increase in oxidative stress markers (PC by 13.3 ± 3.7%; TBARS by 7.2 ± 2.7%, p<0.01) and CK activity, despite the fact that total work done was 10.9 ± 3.6% greater in the post- compared with the pre-training exercise test. Training also induced a marked elevation of antioxidant status indices (TAC by 38.4 ± 7.2%; CAT by 26.2 ± 10.1%; GPX by 3.0 ± 0.6%, p<0.01). Short-term HIT attenuates oxidative stress and up-regulates antioxidant activity after only nine training sessions totaling 22 min of high intensity exercise, further supporting its positive effect not only on physical conditioning but also on health promotion.

Cytokines in muscle damage.

Multiple cellular and molecular processes are rapidly activated following skeletal muscle damage to restore normal muscle structure and function. These processes typically involve an inflammatory response and potentially the consequent occurrence of secondary damage before their resolution and the completion of muscle repair or regeneration. The overall outcome of the inflammatory process is potentially divergent, with the induction of prolonged inflammation and further muscle damage, or its active termination and the promotion of muscle repair and regeneration. The final, detrimental, or beneficial effect of the inflammatory response on muscle repair is influenced by specific interactions between inflammatory and muscle cell-derived cytokines that act as positive and/or negative regulators to coordinate local and systemic inflammatory-related events and modulate muscle repair process. A crucial balance between proinflammatory and anti-inflammatory cytokines appears to attenuate an excessive inflammatory reaction, prevent the development of muscle fibrosis, and adequately promote the regenerative process. In this review, we address the interactive cytokine responses following muscle damage, in the context of induction and progression, or resolution of muscle inflammation and the promotion of muscle repair.

Preferential expression of IGF-1Ec (MGF) transcript in cancerous tissues of human prostate: evidence for a novel and autonomous growth factor activity of MGF E peptide in human prostate cancer cells.

By alternative splicing the IGF‐1 gene produces several different transcripts, including IGF‐1Ec (MGF). The latter has been mainly associated with muscle regeneration processes.

Systemic cytokine response following exercise-induced muscle damage in humans

Abstract Background: Muscle adaptation which occurs following eccentric exercise-induced muscle damage has been associated with an acute inflammatory response. The purpose of this study was to investigate serum interleukin-6 (IL-6), osteoprotegerin and receptor activator of nuclear factor kB ligand (OPG/RANKL) concentrations following muscle damage. We measured changes for several days following muscle damage. Methods: Ten healthy young males performed an eccentric exercise protocol using their quadriceps. Blood samples were withdrawn before and at 6 h, 2 days, 5 days and 16 days post-exercise. Functional and clinical measurements were performed before, and on days 1, 2, 5, 8, 12 and 16 post-exercise. Results: The exercise protocol resulted in muscle damage, indicated by changes in biochemical markers. An increase in IL-6 and OPG, and a decrease in RANKL concentrations were seen at 6 h and on day 2 post-exercise; the OPG:RANKL ratio was increased at 6 h post-exercise (p<0.05). Conclusions: Changes in IL-6 and OPG/RANKL system may represent systemic responses in muscle inflammation and repair processes. However, further studies are needed to elucidate a potential systemic and/or local role of the OPG/RANKL system in skeletal muscle repair. Clin Chem Lab Med 2009;47:777–82.

The complexity of the IGF1 gene splicing, posttranslational modification and bioactivity.

The insulinlike growth factor-I (IGF-I) is an important factor which regulates a variety of cellular responses in multiple biological systems. The IGF1 gene comprises a highly conserved sequence and contains six exons, which give rise to heterogeneous mRNA transcripts by a combination of multiple transcription initiation sites and alternative splicing. These multiple transcripts code for different precursor IGF-I polypeptides, namely the IGF-IEa, IGF-IEb and IGF-IEc isoforms in humans, which also undergo posttranslational modifications, such as proteolytic processing and glycosylation. IGF-I actions are mediated through its binding to several cell-membrane receptors and the IGF-I domain responsible for the receptor binding is the bioactive mature IGF-I peptide, which is derived after the posttranslational cleavage of the pro-IGF-I isoforms and the removal of their carboxy-terminal E-peptides (that is, the Ea, Eb and Ec). Interestingly, differential biological activities have been reported for the different IGF-I isoforms, or for their E-peptides, implying that IGF-I peptides other than the IGF-I ligand also possess bioactivity and, thus, both common and unique or complementary pathways exist for the IGF-I isoforms to promote biological effects. The multiple peptides derived from IGF-I and the differential expression of its various transcripts in different conditions and pathologies appear to be compatible with the distinct cellular responses observed to the different IGF-I peptides and with the concept of a complex and possibly isoform-specific IGF-I bioactivity. This concept is discussed in the present review, in the context of the broad range of modifications that this growth factor undergoes which might regulate its mechanism(s) of action.

Angle-specific impairment of elbow flexors strength after isometric exercise at long muscle length

Abstract In this study, we examined the long-term reductions in maximal isometric force (MIF) caused by a protocol of repeated maximal isometric contractions at long muscle length. Furthermore, we wished to ascertain whether the reductions in MIF are dependent on muscle length — that is, are the reductions in MIF more pronounced when the muscle contracts at a short length. The MIF of the elbow flexors of seven young male volunteers was measured at five different elbow angles between 50° and 160°. On a separate day, the participants performed 50 maximal voluntary isometric muscle contractions with the elbow flexors at a lengthened positions that is, with the shoulder hyperextended at 45° and the elbow joint fixed at 140°. Following this exercise, the MIF at the five elbow angles, range of motion, muscle soreness and plasma creatine kinase activity were measured at 24 h intervals for 4 days. On day 1, the decline in MIF was higher at the more acute elbow angles of 50° (42±8%) and 70° (39±8%; both P<0.01) than at 90° (26±4%) and 140° (16±3%; both P<0.01). No significant reduction in MIF was evident at an elbow angle of 160°. Maximal isometric force at an elbow angle of 140° was fully restored on day 3, whereas at an angle of 50° it remained depressed for the 4 day observation period. Restoration of MIF was a function of the elbow angle, with force recovery being less at the smaller angles. The range of motion was decreased by 14±2° on day 1 (P<0.01) and did not return to baseline values by day 4. Muscle soreness ratings remained significantly elevated for the 4 day period. Serum creatine kinase peaked on day 1 (522±129 IU, P<0.01) and decreased thereafter. We conclude that the disproportionate decrease in MIF at the small elbow angles and the length-specific recovery in MIF after repeated maximal isometric contractions at long muscle length may be explained by the presence of overstretched sarcomeres that increased in series compliance of the muscle, therefore causing a rightward shift of the force-length relationship.

Optimizing IGF-I for skeletal muscle therapeutics

It is virtually undisputed that IGF-I promotes cell growth and survival. However, the presence of several IGF-I isoforms, vast numbers of intracellular signaling components, and multiple receptors results in a complex and highly regulated system by which IGF-I actions are mediated. IGF-I has long been recognized as one of the critical factors for coordinating muscle growth, enhancing muscle repair, and increasing muscle mass and strength. How to optimize this panoply of pathways to drive anabolic processes in muscle as opposed to aberrant growth in other tissues is an area that deserves focus. This review will address how advances in the bioavailability, potency, and tissue response of IGF-I can provide new potential directions for skeletal muscle therapeutics.

Evidence for the Possible Biological Significance of the igf-1 Gene Alternative Splicing in Prostate Cancer.

Insulin-like growth factor-I (IGF-I) has been implicated in the pathogenesis of prostate cancer (PCa), since it plays a key role in cell proliferation, differentiation, and apoptosis. The IGF-I actions are mediated mainly via its binding to the type I IGF receptor (IGF-IR), however IGF-I signaling via insulin receptor (IR) and hybrid IGF-I/IR is also evident. Different IGF-I mRNA splice variants, namely IGF-IEa, IGF-IEb, and IGF-IEc, are expressed in human cells and tissues. These transcripts encode several IGF-I precursor proteins which contain the same bioactive product (mature IGF-I), however, they differ by the length of their signal peptides on the amino-terminal end and the structure of the extension peptides (E-peptides) on the carboxy-terminal end. There is an increasing interest in the possible different role of the IGF-I transcripts and their respective non-(mature)IGF-I products in the regulation of distinct biological activities. Moreover, there is strong evidence of a differential expression profile of the IGF-I splice variants in normal versus PCa tissues and PCa cells, implying that the expression pattern of the various IGF-I transcripts and their respective protein products may possess different functions in cancer biology. Herein, the evidence that the IGF-IEc transcript regulates PCa growth via Ec peptide specific and IGF-IR/IR-independent signaling is discussed.

Physical Exercise Positively Influences Breast Cancer Evolution

Breast cancer is one of the most commonly diagnosed types of cancer in women. Its pathogenesis involves genetic, hormonal, and environmental factors. A large body of evidence indicates that physical activity has positive effects on every aspect of breast cancer evolution, including prevention, medical treatment, and aftercare clinical settings. Thus, different types of exercise can influence the prevention and progression of the disease through several common mechanisms, such as reduction of insulin resistance and improvement of immunity and cardiovascular function. Furthermore, acute and chronic symptoms of breast cancer, such as cachexia, muscle mass loss, fatigue, cardiotoxicity, weight gain, hormone alterations, bone loss, and psychologic adverse effects, may all be favorably influenced by regular exercise. We review the relation of intensity and duration of exercise with potential pathophysiologic pathways, including obesity-related hormones and sex steroid hormone production, oxidative stress, epigenetic alterations such as DNA hypomethylation, and changes in telomere length, within the context of the beneficial effects of exercise. The potential role of exercise in reducing the intensity of the adverse effects that result from breast cancer and anticancer treatment is also discussed.