Identification of Proteomic Biomarkers in Proliferative Verrucous Leukoplakia through Liquid Chromatography With Tandem Mass Spectrometry.

Proliferative verrucous leukoplakia (PVL) is an oral potentially malignant disorder associated with high risk of malignant transformation. Currently, there is no treatment available, and restrictive follow-up of patients is crucial for a better prognosis. Oral leukoplakia (OL) shares some clinical and microscopic features with PVL but exhibits different clinical manifestations and a lower rate of malignant transformation. This study aimed to investigate the proteomic profile of PVL in tissue and saliva samples to identify potential diagnostic biomarkers with therapeutic implications. Tissue and saliva samples obtained from patients with PVL were compared with those from patients with oral OL and controls. Label-free liquid chromatography with tandem mass spectrometry was employed, followed by qualitative and quantitative analyses, to identify differentially expressed proteins. Potential biomarkers were identified and further validated using immunohistochemistry. Staining intensity scan analyses were performed on tissue samples from patients with PVL, patients with OL, and controls from Brazil, Spain, and Finland. The study revealed differences in the immune system, cell cycle, DNA regulation, apoptosis pathways, and the whole proteome of PVL samples. In addition, liquid chromatography with tandem mass spectrometry analyses showed that calreticulin (CALR), receptor of activated protein C kinase 1 (RACK1), and 14-3-3 Tau-protein (YWHAQ) were highly expressed in PVL samples. Immunohistochemistry validation confirmed increased CARL expression in PVL compared with OL. Conversely, RACK1 and YWHA were highly expressed in oral potentially malignant disorder compared to the control group. Furthermore, significant differences in CALR and RACK1 expression were observed in the OL group when comparing samples with and without oral epithelial dysplasia, unlike the PVL. This research provides insights into the molecular mechanisms underlying these conditions and highlights potential targets for future diagnostic and therapeutic approaches.

Protein-based salivary biomarkers for the diagnosis of periodontal diseases: Systematic review and meta-analysis.

Objective: This systematic review and meta-analysis was aimed at determining differentially expressed protein-based biomarkers detectable in the saliva for the diagnosis of major periodontal diseases. Methods: A literature review was conducted through January 31, 2022. The methodological quality and risk of bias were assessed with the Newcastle-Ottawa scale for case-control studies. Heterogeneity among studies was analysed with the Q statistical test and the I2 test. p-values lower than 0.10 and I2 values higher than 50% indicated high heterogeneity among studies; therefore, the random-effects model was used. The analysis of biological pathways associated with the differentially expressed protein markers was performed with the STITCH integration analysis tool and was limited to interactions with high confidence levels (0.7). Results: Of all protein-based biomarkers detected, 12 were suitable for meta-analysis: IL-1ß, MIP-1α, albumin, TNF-α, ICTP, Ig-A, lactoferrin, MMP-8, IL-6, IL-8, IL-17 and PGE2. The salivary markers with high applicability were IL-1ß for differentiating patients with chronic periodontal disease from patients with gingivitis with an OE = 73.5 pg/mL; ICTP for differentiating patients with chronic periodontal disease from healthy control patients with an OE = 0.091 ng/mL; and PGE2 for differentiating patients with chronic periodontal disease from healthy control patients with an OE = 36.3 pg/mL. Conclusions: The biomarkers with the highest differential expression and the greatest potential for clinical applicability are IL-1ß for differentiating periodontitis from gingivitis, and ICTP and PGE2 for differentiating periodontitis from healthy status.

Effects of Ferroptosis on the Metabolome in Cardiac Cells: The Role of Glutaminolysis.

Ferroptosis is a novel iron-dependent regulated cell death mechanism that affects cell metabolism; however, a detailed metabolomic analysis of ferroptotic cells is not yet available. Here, we elucidated the metabolome of H9c2 cardioblasts by gas chromatography-mass spectrometry during ferroptosis induced by RSL3, a GPX4 inhibitor, in the presence of ferrostatin-1 (a ferroptosis inhibitor), XJB-5-131 (a mitochondrial-targeted ROS scavenger), or TSM-1005-44 (a newly developed cellular ROS scavenger). Results demonstrated that RSL3 decreased the levels of amino acids involved in glutathione synthesis more than two-fold. In contrast, saturated fatty acids levels were markedly increased in RSL3-challenged cells, with no effects on unsaturated fatty acids. RSL3 significantly altered the levels of mitochondrial tricarboxylic acid cycle intermediates; isocitrate and 2-oxoglutarate were found to increase, whereas succinate was significantly decreased in RSL3-challenged cells. Ferrostatin-1, XJB-5-131, and TSM-1005-44 prevented RSL3-induced cell death and conserved the metabolomic profile of the cells. Since 2-oxoglutarate is involved in the regulation of ferroptosis, particularly through glutamine metabolism, we further assessed the role of glutaminolysis in ferroptosis in H9c2 cardioblasts. Genetic silencing of GLS1, which encodes the K-type mitochondrial glutaminase (glutaminase C), protected against ferroptosis in the early stage. In conclusion, our study demonstrates that RSL3-induced ferroptosis impairs the metabolome of H9c2 cardioblasts.

Usefulness of protein-based salivary markers in the diagnosis of oral potentially malignant disorders: A systematic review and meta-analysis.

By using a meta-analytical approach, this study aimed to analyse the diagnostic capacity of protein-based biomarkers in saliva for the differential diagnosis of oral potentially malignant disorders (OPMDs) and oral squamous cell carcinoma (OSCC) from healthy individuals as control group (HCG).Articles on protein-based biomarkers in saliva, which provided quantitative expression in individuals with clinical and histopathological diagnosis of OPMD or oral leukoplakia (OL) were considered eligible. Searches were conducted in eight electronic databases. The methodological quality was assessed using the Quality Assessment of Diagnostic Studies tool (QUADAS-2). Functional analysis was also performed. Meta-analyses were performed using the OpenMeta tool (Analyst).Meta-analysis was possible for 4 of the 11 biomarkers studied. Only the carcinoembryonic antigen (CEA) and the soluble fragment of cytokeratin 19 (CYFRA21) were significant for the OSCC/OPMD subgroup, both with a very low heterogeneity. CEA had an OE = 25.854 (CI95%: 13.215-38.492, p< 0.001, I2 = 0) and CYFRA21 had an OE = 9.317 (CI95%: 9.014-9.619, p< 0.001, I2 = 0). For the OPMD/HCG subgroup, only CYFRA21 was significant, with an OE = 3.679 (CI95%: 0.663-6.696, p= 0.017) although with high heterogeneity (I2 = 91.24).The CEA and CYFRA21 markers proved very useful when differentiating OSCC from OPMD. The CYFRA21 was the only protein that was capable of distinguishing between OPMD and healthy controls.

Distribución de la adiposidad en nadadores según categorías de pubertad / Body fat distribution in young swimmers according to pubertal development

La distribución de la adiposidad al igual que otros componentes del físico humano, experimenta cambios a lo largo de la ontogenia. El objetivo de esta investigación fue analizar la distribución de la adiposidad en 178 nadadores venezolanos, clasificados por maduración sexual (prepúber, púber inicial y púber avanzado). Se utilizó el análisis de componentes principales (ACP), a partir de las regresiones lineales de los logaritmos de ocho pliegues de tejido adiposo. En los nadadores se identificaron 3 ACP para el prepúber y púber avanzado y 2 en el inicial. El Primer Componente (PC1) en el prepúber está asociado con la adiposidad total; en inicial y avanzado, indica un patrón de distribución tronco-extremidad que compromete a todos los pliegues del tronco tanto superior como inferior. En las nadadoras se identificaron 2 componentes en el prepúber y 3 en inicial y avanzado, en estas últimas se acumula el mayor porcentaje de varianza (81.5%). Los PC1 en los tres grados de maduración están asociados con la adiposidad total, el Segundo Componente (PC2) indica la distribución tronco-extremidad. La agrupación por categorías de maduración es bastante consistente ya que el patrón de dispersión de las edades cronológicas no genera una variabilidad considerable. La centralidad en los varones se corresponde con la misma tendencia observada en la población no atlética venezolana. Finalmente se establece la necesidad de considerar como variable pivotal cada uno de los estadios de maduración cuya importancia se ha señalado en la detección de talentos de niños y adolescentes.

[Anthropometric characteristics and skeletal maturity of male Venezuelan swimmers]. / Caracterización antropométrica y maduración ósea de nadadores venezolanos.

Knowledge of the morph-physiological conditions of young athletes, it of the utmost importance for planning better trainning programs and to identify those characteristics that lead to a better performance. This paper aims to classify a group of 114 male Venezuelan swimmers of the Miranda State contingent, aged between 7.00 and 18.99 years old, based on their anthropometric characteristics and skeletal maturity. For this purpose multivariate methods: Analysis of Principal Components (APC) and the Automatic Classification were employed. Anthropometric variables followed the International Society for the Advancement of Kinantropometry (ISAK) guidelines. Skeletal maturity and five indexes of body composition: brachial, crural, cormic and acromiale-iliac indexes, were additionally considered in the analysis. Three groups emerged very well defined and homogenous within and between groups. The first group identifies the younger swimmers with the smallest values in corporal dimensions. The second group comprises those swimmers characterized fundamentally by a greater development of the bicrestal diameter. In the third group, greater values of the anthropometrical variables were found, that shaped the typical profile of the swimmers, mainly characterized by a trapezoidal trunk. Most of the swimmers are advanced in skeletal age related to chronological age. This clustering of the swimmers permits a suitable way to identify the bio-morphological characteristics of the athletes.

Caracterización antropométrica y maduración ósea de nadadores venezolanos / Anthropometric characteristics and skeletal maturity of male Venezuelan swimmers

El conocimiento de las condiciones morfo-fisiológicas de los jóvenes atletas, es fundamental para asignar dosificaciones correctas de las cargas de entrenamientos y obtener mejores resultados en las competencias sin afectar la salud de los deportistas. La presente investigación se propone clasificar a un grupo de nadadores masculinos del Estado Miranda en función de sus características antropométricas y de maduración ósea, usando métodos multivariantes: Análisis de Componentes Principales (ACP) y la Clasificación Automática. Fueron evaluados 114 nadadores con edades comprendidas entre 7,00 y 18,99 años. Se tomaron 12 variables antropométricas siguiendo los lineamientos de la Sociedad Internacional para el Avance de la Kinantropometría (ISAK), y se consideraron la Edad Ósea, Composición Corporal en 5 componentes e índices Braquial (IB), Crural (IC); Córmico o Esquelético, y el Acromio-Iliaco. Se obtuvieron tres grupos bien definidos y homogéneos entre sí. En el primero están ubicados los nadadores con menores valores, tanto en edad como en dimensiones corporales. En el segundo, aquellos que se encuentran en una etapa intermedia caracterizados fundamentalmente por un marcado desarrollo del diámetro bicrestal. En el tercer grupo se localizan los de mayores valores, los cuales mostraron la forma típica del nadador de tronco trapezoidal. La mayoría de los nadadores presentaron una edad ósea adelantada respecto a su edad cronológica. Esta agrupación de los nadadores resultó una buena guía para determinar las características bio-morfológicas de los atletas