Melanoma in the liver: Oxidative stress and the mechanisms of metastatic cell survival.

Metastatic melanoma is a fatal disease with a rapid systemic dissemination. The most frequent target sites are the liver, bone, and brain. Melanoma metastases represent a heterogeneous cell population, which associates with genomic instability and resistance to therapy. Interaction of melanoma cells with the hepatic sinusoidal endothelium initiates a signaling cascade involving cytokines, growth factors, bioactive lipids, and reactive oxygen and nitrogen species produced by the cancer cell, the endothelium, and also by different immune cells. Endothelial cell-derived NO and H2O2 and the action of immune cells cause the death of most melanoma cells that reach the hepatic microvascularization. Surviving melanoma cells attached to the endothelium of pre-capillary arterioles or sinusoids may follow two mechanisms of extravasation: a) migration through vessel fenestrae or b) intravascular proliferation followed by vessel rupture and microinflammation. Invading melanoma cells first form micrometastases within the normal lobular hepatic architecture via a mechanism regulated by cross-talk with the stroma and multiple microenvironment-related molecular signals. In this review special emphasis is placed on neuroendocrine (systemic) mechanisms as potential promoters of liver metastatic growth. Growing metastatic cells undergo functional and metabolic changes that increase their capacity to withstand oxidative/nitrosative stress, which favors their survival. This adaptive process also involves upregulation of Bcl-2-related antideath mechanisms, which seems to lead to the generation of more resistant cell subclones.

Nicotinamide Riboside and Pterostilbene Cooperatively Delay Motor Neuron Failure in ALS SOD1G93A Mice.

Oxidative stress-induced damage is a major mechanism in the pathophysiology of amyotrophic lateral sclerosis (ALS). A recent human clinical trial showed that the combination of nicotinamide riboside (NR) and pterostilbene (PT), molecules with potential to interfere in that mechanism, was efficacious in ALS patients. We examined the effect of these molecules in SOD1G93A transgenic mice, a well-stablished model of ALS. Assessment of neuromotor activity and coordination was correlated with histopathology, and measurement of proinflammatory cytokines in the cerebrospinal fluid. Cell death, Nrf2- and redox-dependent enzymes and metabolites, and sirtuin activities were studied in isolated motor neurons. NR and PT increased survival and ameliorated ALS-associated loss of neuromotor functions in SOD1G93A transgenic mice. NR and PT also decreased the microgliosis and astrogliosis associated with ALS progression. Increased levels of proinflammatory cytokines were observed in the cerebrospinal fluid of mice and humans with ALS. NR and PT ameliorated TNFα-induced oxidative stress and motor neuron death in vitro. Our results support the involvement of oxidative stress, specific Nrf2-dependent antioxidant defenses, and sirtuins in the pathophysiology of ALS. NR and PT interfere with the mechanisms leading to the release of proapoptotic molecular signals by mitochondria, and also promote mitophagy.

Adding the Load Just Above Sticking Point Using Elastic Bands Optimizes Squat Performance, Perceived Effort Rate, and Cardiovascular Responses.

Modifying basal elongation of elastic bands (EB) has been proven useful to increase some parameters of the intensity in variable resistance training. Therefore, the question arises as to whether the pertinent resistance could be applied with EB immediately above the sticking point in squat exercises to optimize the performance. The purpose was to analyze some variables of the external (kilograms and number of repetitions) and internal load (heart rate, blood pressure, and rate of perceived exertion) after six different conditions of the squat exercise when using weight plates (WP) or EB (placed at different points of the range of motion) and applying maximal or submaximal effort. Twenty physically active males (25.50 ± 5.26 yrs) underwent two sessions for familiarization and one for assessment. The six conditions (three with WP and three with EB) were randomly performed. The sticking point of each subject was measured using the knee joint angle and the resistance was applied with EB at this height. Immediately after finishing each set subjects reported perceived effort rate and cardiovascular measurements were taken. Repetitions completed, and kilograms used were recorded. Repeated measures testing evaluated differences between conditions. EB permitted performing 8 more repetitions compared to WP when the same load was added at standing position. Adding the load immediately above the sticking point significantly (p < 0.05) increased 24.7% the kilograms used and permitted participants to perform 3 more repetitions. Internal load measurements suggested that EB could significantly (p < 0.05) reduce the perceived effort rate and/or physiological stress depending on their application. EB are a suitable device to load the bar for squat exercises in fit young men. According to the necessities of the subjects, if the load with EB is added at different points of the range of motion, it could be possible to overcome the sticking point, to maximize the performance and/or modulate cardiovascular and perceptual responses.

Radioprotection and Radiomitigation: From the Bench to Clinical Practice.

The development of protective agents against harmful radiations has been a subject of investigation for decades. However, effective (ideal) radioprotectors and radiomitigators remain an unsolved problem. Because ionizing radiation-induced cellular damage is primarily attributed to free radicals, radical scavengers are promising as potential radioprotectors. Early development of such agents focused on thiol synthetic compounds, e.g., amifostine (2-(3-aminopropylamino) ethylsulfanylphosphonic acid), approved as a radioprotector by the Food and Drug Administration (FDA, USA) but for limited clinical indications and not for nonclinical uses. To date, no new chemical entity has been approved by the FDA as a radiation countermeasure for acute radiation syndrome (ARS). All FDA-approved radiation countermeasures (filgrastim, a recombinant DNA form of the naturally occurring granulocyte colony-stimulating factor, G-CSF; pegfilgrastim, a PEGylated form of the recombinant human G-CSF; sargramostim, a recombinant granulocyte macrophage colony-stimulating factor, GM-CSF) are classified as radiomitigators. No radioprotector that can be administered prior to exposure has been approved for ARS. This differentiates radioprotectors (reduce direct damage caused by radiation) and radiomitigators (minimize toxicity even after radiation has been delivered). Molecules under development with the aim of reaching clinical practice and other nonclinical applications are discussed. Assays to evaluate the biological effects of ionizing radiations are also analyzed.

Oxidative Stress, Neuroinflammation and Mitochondria in the Pathophysiology of Amyotrophic Lateral Sclerosis.

Amyotrophic lateral sclerosis (ALS) is a progressive motor neuron (MN) disease. Its primary cause remains elusive, although a combination of different causal factors cannot be ruled out. There is no cure, and prognosis is poor. Most patients with ALS die due to disease-related complications, such as respiratory failure, within three years of diagnosis. While the underlying mechanisms are unclear, different cell types (microglia, astrocytes, macrophages and T cell subsets) appear to play key roles in the pathophysiology of the disease. Neuroinflammation and oxidative stress pave the way leading to neurodegeneration and MN death. ALS-associated mitochondrial dysfunction occurs at different levels, and these organelles are involved in the mechanism of MN death. Molecular and cellular interactions are presented here as a sequential cascade of events. Based on our present knowledge, the discussion leads to the idea that feasible therapeutic strategies should focus in interfering with the pathophysiology of the disease at different steps.

Efficacy and tolerability of EH301 for amyotrophic lateral sclerosis: a randomized, double-blind, placebo-controlled human pilot study.

BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease, characterized by progressive loss of spinal and cortical motor neurons, leading to muscular atrophy, respiratory failure, and ultimately death. There is no known cure, and the clinical benefit of the two drugs approved to treat ALS remains unclear. Novel disease-modifying therapeutics that are able to modulate the disease course are desperately needed. Our objective was to evaluate the efficacy and tolerability of Elysium Health's candidate drug EH301 in people with ALS (PALS). METHODS: This was a single-center, prospective, double-blind, randomized, placebo-controlled pilot study. Thirty-two PALS were recruited thanks to the collaboration of the Spanish Foundation for ALS Research (FUNDELA). Study participants were randomized to receive either EH301 or placebo and underwent evaluation for 4 months. Differences between EH301 and placebo-treated participants were evaluated based on standard clinical endpoints, including the revised ALS functional rating scale (ALSFRS-R), forced vital capacity (FVC), and the Medical Research Council (MRC) grading scale. RESULTS: Compared to placebo, participants treated with EH301 demonstrated significant improvements in the ALSFRS-R score, pulmonary function, muscular strength, and in skeletal muscle/fat weight ratio. EH301 was shown to significantly slow the progression of ALS relative to placebo, and even showed improvements in several key outcome measures compared with baseline. CONCLUSIONS: This study provides evidence in support of the disease-modifying effects of EH301 for the treatment of ALS.

Oxidative stress and antioxidants in the pathophysiology of malignant melanoma.

The high number of somatic mutations in the melanoma genome associated with cumulative ultra violet (UV) exposure has rendered it one of the most difficult of cancers to treat. With new treatment approaches based on targeted and immune therapies, drug resistance has appeared as a consistent problem. Redox biology, including reactive oxygen and nitrogen species (ROS and RNS), plays a central role in all aspects of melanoma pathophysiology, from initiation to progression and to metastatic cells. The involvement of melanin production and UV radiation in ROS/RNS generation has rendered the melanocytic lineage a unique system for studying redox biology. Overall, an elevated oxidative status has been associated with melanoma, thus much effort has been expended to prevent or treat melanoma using antioxidants which are expected to counteract oxidative stress. The consequence of this redox-rebalance seems to be two-fold: on the one hand, cells may behave less aggressively or even undergo apoptosis; on the other hand, cells may survive better after being disseminated into the circulating system or after drug treatment, thus resulting in metastasis promotion or further drug resistance. In this review we summarize the current understanding of redox signaling in melanoma at cellular and systemic levels and discuss the experimental and potential clinic use of antioxidants and new epigenetic redox modifiers.

Glucocorticoid receptor antagonism overcomes resistance to BRAF inhibition in BRAFV600E-mutated metastatic melanoma.

Clinical applications of glucocorticoids (GC) in Oncology are dependent on their pro-apoptotic action to treat lymphoproliferative cancers, and to alleviate side effects induced by chemotherapy and/or radiotherapy. However, the mechanism(s) by which GC may also promote tumor progression remains unclear. GC receptor (GR) knockdown decreases the antioxidant protection of highly metastatic B16-F10 melanoma cells. We hypothesize that a GR antagonist (RU486, mifepristone) could increase the efficacy of BRAF-related therapy in BRAFV600E-mutated metastatic melanoma. In vivo formed spontaneous skin tumors were reinoculated into nude mice to expand the metastases of different human BRAFV600E melanoma cells. The GR content of melanoma cell lines was measured by [3H]-labeled ligand binding assay. Nuclear Nrf2 and its transcription activity was investigated by RT-PCR, western blotting, and by measuring Nrf2- and redox state-related enzyme activities and metabolites. GR knockdown was achieved using lentivirus, and GR overexpression by transfection with the NR3C1 plasmid. shRNA-induced selective Bcl-xL, Mcl-1, AKT1 or NF-κB/p65 depletion was used to test the efficacy of vemurafenib (VMF) and RU486 against BRAFV600E-mutated metastatic melanoma. During early progression of skin melanoma metastases, RU486 and VMF induced a drastic metastases regression. However, treatment at an advanced stage of growth demonstrated the development of resistance to RU486 and VMF. This resistance was mechanistically linked to overexpression of specific proteins of the Bcl-2 family (Bcl-xL and Mcl-1 in our experimental models). We found that melanoma resistance is decreased if AKT and NF-κB signaling pathways are blocked. Our results highlight mechanisms by which metastatic melanoma cells adapt to survive.

Polyphenolic Phytochemicals in Cancer Prevention and Therapy: Bioavailability versus Bioefficacy.

Natural polyphenols are organic chemicals which contain phenol units in their structures. They show antitumor properties. However, a key problem is their short half-life and low bioavailability under in vivo conditions. Still, definitively demonstrating the human benefits of isolated polyphenolic compounds (alone or in combination) using modern scientific methodology has proved challenging. The most common discrepancy between experimental and clinical observations is the use of nonphysiologically relevant concentrations of polyphenols in mechanistic studies. Thus, it remains highly controversial how applicable underlying mechanisms are with bioavailable concentrations and biological half-life. The present Perspective analyses proposed antitumor mechanisms, in vivo reported antitumor effects, and possible mechanisms that may explain discrepancies between bioavailability and bioefficacy. Polyphenol metabolism and possible toxic side effects are also considered. Our main conclusion emphasizes that these natural molecules (and their chemical derivatives) indeed can be very useful, not only as cancer chemopreventive agents but also in oncotherapy.

A New Equation for Predicting Evolution of Oral Pain in Orthodontic Treatment: A Longitudinal, Prospective Cohort Study.

AIMS: To develop an equation capable of relating the evolution of oral pain to the time elapsed, measured from the moment of dental archwire fitting and identifying when pain begins, peaks, and ends; and secondly, to compare pain during orthodontic treatment in relation to archwire material (steel or nickel-titanium [Ni-Ti]) and position (maxillary or mandibular) and patient age (child, teenager, or adult) and gender (male or female). METHODS: A longitudinal prospective cohort study was conducted of 112 patients who filled in a scale to evaluate pain, noting the times when the pain occurred. The total sample consisted of 60 males and 52 females with a mean (± standard deviation [SD]) age of 19.8 ± 6.2 years. The sample was divided into five groups depending on archwire material and position, and patient age and gender. A univariate four-way ANOVA model was performed to compare mean pain levels between groups. Bonferroni test was used for multiple comparisons. A univariate nonlinear regression model was carried out for pain level, 95% confidence intervals (95% CI) were calculated, and the statistic R² was used. RESULTS: An equation was developed based on pain levels in relation to time elapsed, measured from the moment when the archwire had been fitted in the mouth. The equation had three coefficients related to mean pain values: overall pain, peak pain, and how pain decreased. It fitted all study groups with a correlation coefficient > 0.9. The model showed that pain levels were influenced by archwire material and patient gender and age, but not archwire position. CONCLUSION: The equation reproduced the data registered and can be applied to studies of pain derived from archwires, and this methodology could be used for other external agents fitted in the mouth. Patients receiving dental treatment involving external agents can be made aware of the pain they can expect to experience. This will enable them to distinguish expected pain from other pain, which will help them identify other pathologies requiring medical attention and to approach treatment with better motivation since the pattern of pain evolution is known in advance.

Pterostilbene Decreases the Antioxidant Defenses of Aggressive Cancer Cells In Vivo: A Physiological Glucocorticoids- and Nrf2-Dependent Mechanism.

AIMS: Polyphenolic phytochemicals have anticancer properties. However, in mechanistic studies, lack of correlation with the bioavailable concentrations is a critical issue. Some reports had suggested that these molecules downregulate the stress response, which may affect growth and the antioxidant protection of malignant cells. Initially, we studied this potential underlying mechanism using different human melanomas (with genetic backgrounds correlating with most melanomas), growing in nude mice as xenografts, and pterostilbene (Pter, a natural dimethoxylated analog of resveratrol). RESULTS: Intravenous administration of Pter decreased human melanoma growth in vivo. However, Pter, at levels measured within the tumors, did not affect melanoma growth in vitro. Pter inhibited pituitary production of the adrenocorticotropin hormone (ACTH), decreased plasma levels of corticosterone, and thereby downregulated the glucocorticoid receptor- and nuclear factor (erythroid-derived 2)-like 2 (Nrf2)-dependent antioxidant defense system in growing melanomas. Exogenous corticosterone or genetically induced Nrf2 overexpression in melanoma cells prevented the inhibition of tumor growth and decreased antioxidant defenses in these malignant cells. These effects and mechanisms were also found in mice bearing different human pancreatic cancers. Glutathione depletion (selected as an antimelanoma strategy) facilitated the complete elimination by chemotherapy of melanoma cells isolated from mice treated with Pter. INNOVATION: Although bioavailability-related limitations may preclude direct anticancer effects in vivo, natural polyphenols may also interfere with the growth and defense of cancer cells by downregulating the pituitary gland-dependent ACTH synthesis. CONCLUSIONS: Pter downregulates glucocorticoid production, thus decreasing the glucocorticoid receptor and Nrf2-dependent signaling/transcription and the antioxidant protection of melanoma and pancreatic cancer cells. Antioxid. Redox Signal. 24, 974-990.

Infrared thermal imaging in the diagnosis of musculoskeletal injuries: a systematic review and meta-analysis.

OBJECTIVE: Musculoskeletal injuries occur frequently. Diagnostic tests using ionizing radiation can lead to problems for patients, and infrared thermal imaging could be useful when diagnosing these injuries. CONCLUSION: A systematic review was performed to determine the diagnostic accuracy of infrared thermal imaging in patients with musculoskeletal injuries. A meta-analysis of three studies evaluating stress fractures was performed and found a lack of support for the usefulness of infrared thermal imaging in musculoskeletal injuries diagnosis.

Effects of endodontic treatment on apparent Young's modulus of human teeth: in vitro study using speckle interferometry.

The introduction of new techniques for endodontic procedures requires the analysis of the biomechanical behavior of dental structures. Digital speckle shearing pattern interferometry (DSSPI) is a nondestructive optical measuring technique that allows one to directly quantify deformations in teeth that are subjected to stress. DSSPI technique was applied to measure small deformations caused by flexion in different types of teeth. The test was carried out both before and after endodontic treatment with the ProTaper method in order to evaluate the variation of dental elasticity, taking into the account the type of tooth and the endodontic treatment. The results obtained show that dental elasticity, established by means of the apparent Young's modulus, before and after the endodontic treatment, differs between incisors and premolars. The endodontic process does not affect dental elasticity (p>0.7). Specifically, 57.1% of central incisors and 56.3% of second premolars slightly increase their elasticity after the endodontic process. In turn, 42.9% of central incisors and 43.7% of second premolars slightly decrease elasticity. The endodontic treatment especially affects the "neutral fibre"; therefore, there is little influence on elasticity by flexion. However, after finishing the process, the channel was restored with material, which can slightly increase tooth elasticity in some cases.

Estudio mediante una técnica no lesiva, basada en la luz estructurada, de la topografia de la espalda de futbolistas / Study of back topography of football players by a nonlesive method based on structured light

Se presenta un método no lesivo basado en la proyección de luz estructurada con código de color para la obtención de la topografía de la superficie de la espalda en deportistas. El método permite la visualización de las asimetrías existentes en las distintas zonas de la espalda, cervical, dorsal y lumbar, tomando como referencia la posición de las cervicales y de los glúteos. La determinación en estas topografías de variables cuantificadoras para caracterizar la deformidad, tanto en el plano frontal como transversal, permite realizar un diagnóstico precoz de la existencia de patologías asociadas con desviación de la columna en los deportistas integrantes de los clubs deportivos, sobre todo en las edades infantiles y juveniles, donde la prevalencia de la escoliosis es mayor. En este trabajo se ha introducido una nueva variable, el gradiente lumbar, que permite identificar la elevación de los glúteos respecto de la cintura, para cuantificar el desarrollo de los glúteos asociado a la práctica deportiva, así como la asimetría de la zona lumbar. Estas variables topográficas han resultado independientes de la altura y edad de los sujetos del estudio y, por tanto, pueden resultar de interés para valorar el efecto del entrenamiento deportivo en la musculatura, tanto a nivel dorsal como lumbar, a lo largo de la evolución del deportista en estudios longitudinales, así como para realizar las oportunas comparaciones entre las distintas actividades deportivas. En nuestro estudio, aplicado aun equipo de fútbol, se ha podido comprobar el mayor número de valores elevados de la variable que mide la deformidad en el plano horizontal en el grupo de edad de los 14 a los 15 años y una ligera variación en función de la posición que ocupan en el campo (AU)

We present a noninvasive method based on structured light projection with color code to obtain the topography of the back surface in athletes. The method allows the display of the asymmetries in the areas of the back, cervical, dorsal and lumbar, with reference to the position of the neck and buttocks. The determination of quantifier variables in these topographies to characterize the deformity in both the frontal and transverse plane allows an early diagnosis of the existence of pathologies associated with curvature of the spine in athletes members of sports clubs, especially in children and youth ages where the prevalence of scoliosis is greater. In this paper we have introduced a new variable, the lumbar gradient, which identifies the elevation of the buttocks on the waist, to quantify the development of the buttocks associated with the sport, and the asymmetry of the lumbar area. These topographic variables were independent of height and age of athletes and therefore may be of interest to assess the effect of sport training in the musculature, both dorsal and lumbar, along of the evolution the athlete and to make appropriate comparisons between different sport activities. In our study, applied to a football team, it has been found the most elevated values of the variable that measures the deformity in the horizontal plane in the age group of 14 to 15 years and a slight variation depending on the playing position (AU)

Quantifier variables of the back surface deformity obtained with a noninvasive structured light method: evaluation of their usefulness in idiopathic scoliosis diagnosis.

New noninvasive techniques, amongst them structured light methods, have been applied to study rachis deformities, providing a way to evaluate external back deformities in the three planes of space. These methods are aimed at reducing the number of radiographic examinations necessary to diagnose and follow-up patients with scoliosis. By projecting a grid over the patient's back, the corresponding software for image treatment provides a topography of the back in a color or gray scale. Visual inspection of back topographic images using this method immediately provides information about back deformity, but it is important to determine quantifier variables of the deformity to establish diagnostic criteria. In this paper, two topographic variables [deformity in the axial plane index (DAPI) and posterior trunk symmetry index (POTSI)] that quantify deformity in two different planes are analyzed. Although other authors have reported the POTSI variable, the DAPI variable proposed in this paper is innovative. The upper normality limit of these variables in a nonpathological group was determined. These two variables have different and complementary diagnostic characteristics, therefore we devised a combined diagnostic criterion: cases with normal DAPI and POTSI (DAPI < or = 3.9% and POTSI < or = 27.5%) were diagnosed as nonpathologic, but cases with high DAPI or POTSI were diagnosed as pathologic. When we used this criterion to analyze all the cases in the sample (56 nonpathologic and 30 with idiopathic scoliosis), we obtained 76.6% sensitivity, 91% specificity, and a positive predictive value of 82%. The interobserver, intraobserver, and interassay variability were studied by determining the variation coefficient. There was good correlation between topographic variables (DAPI and POTSI) and clinical variables (Cobb's angle and vertebral rotation angle).