GPER binding site detection and description: A flavonoid-based docking and molecular dynamics simulations study.

Flavonoids, a phenolic compounds class widely distributed in the plant kingdom, have attracted much interest for their implications on several health and disease processes. Usually, the consumption of this type of compounds is approximately 1 g/d, primarily obtained from cereals, chocolate, and dry legumes ensuring its beneficial role in maintaining the homeostasis of the human body. In this context, in cancer disease prominent data points to the role of flavonoids as adjuvant treatment aimed at the regression of the disease. GPER, an estrogen receptor on the cell surface, has been postulated as a probable orchestrator of the beneficial effects of several flavonoids through modulation/inhibition of various mechanisms that lead to cancer progression. Therefore, applying pocket and cavity protein detection and docking and molecular dynamics simulations (MD), we generate, from a cluster composed of 39 flavonoids, crucial insights into the potential role as GPER ligands, of Puerarin, Isoquercetin, Kaempferol 3-O-glucoside and Petunidin 3-O-glucoside, aglycones whose sugar moiety delimits a new described sugar-acceptor sub-cavity into the cavity binding site on the receptor, as well as of the probable activation mechanism of the receptor and the pivotal residues involved in it. Altogether, our results shed light on the potential use of the aforementioned flavonoids as GPER ligands and for further evaluations in in vitro and in vivo assays to elucidate their probable anti-cancer activity.

Untargeted LC-MS/MS Metabolomics Study of HO-AAVPA and VPA on Breast Cancer Cell Lines.

Breast cancer (BC) is one of the biggest health problems worldwide, characterized by intricate metabolic and biochemical complexities stemming from pronounced variations across dysregulated molecular pathways. If BC is not diagnosed early, complications may lead to death. Thus, the pursuit of novel therapeutic avenues persists, notably focusing on epigenetic pathways such as histone deacetylases (HDACs). The compound N-(2-hydroxyphenyl)-2-propylpentanamide (HO-AAVPA), a derivative of valproic acid (VPA), has emerged as a promising candidate warranting pre-clinical investigation. HO-AAVPA is an HDAC inhibitor with antiproliferative effects on BC, but its molecular mechanism has yet to be deciphered. Furthermore, in the present study, we determined the metabolomic effects of HO-AAVPA and VPA on cells of luminal breast cancer (MCF-7) and triple-negative breast cancer (MDA-MB-231) subtypes. The LC-MS untargeted metabolomic study allowed for the simultaneous measurement of multiple metabolites and pathways, identifying that both compounds affect glycerophospholipid and sphingolipid metabolism in the MCF-7 and MDA-MB-231 cell lines, suggesting that other biological targets were different from HDACs. In addition, there are different dysregulate metabolites, possibly due to the physicochemical differences between HO-AAVPA and VPA.

PAMAM-G4 protect the N-(2-hydroxyphenyl)-2-propylpentanamide (HO-AAVPA) and maintain its antiproliferative effects on MCF-7.

Our work group designed and synthesized a promissory compound N-(2-hydroxyphenyl)-2-propylpentanamide (HO-AAVPA). The HO-AAVPA is a HDAC1 inhibitor and antiproliferative in cancer cell lines. However, HO-AAVPA is poor water solubility and enzymatically metabolized. In this work, the fourth-generation poly(amidoamine) dendrimer (PAMAM-G4) was used as a drug deliver carrier of HO-AAVPA. Moreover, HO-AAVPA and HO-AAVPA-PAMAM complex were submitted to forced degradation studies (heat, acid, base, oxidation and sunlight). Also, the HO-AAVPA-PAMAM-G4 complex was assayed as antiproliferative in a breast cancer cell line (MCF-7). The HO-AAVPA-PAMAM-G4 complex was obtained by docking and experimentally using three pH conditions: acid (pH = 3.0), neutral (pH = 7.0) and basic (pH = 9.0) showing that PAMAM-G4 captureand protect the HO-AAVPA from forced degradation, it is due to sunlight yielded a by-product from HO-AAVPA. In addition, the PAMAM-G4 favored the HO-AAVPA water solubility under basic and neutral pH conditions with significant difference (F(2,18) = 259.9, p < 0.001) between the slopes of the three conditions being the basic condition which solubilizes the greatest amount of HO-AAVPA. Finally, the HO-AAVPA-PAMAM-G4 complex showed better antiproliferative effects on MCF-7 (IC50 = 75.3 µM) than HO-AAVPA (IC50 = 192 µM). These results evidence that PAMAM-G4 complex improve the biological effects of HO-AAVPA.

LC-MS Based Lipidomics Depict Phosphatidylethanolamine as Biomarkers of TNBC MDA-MB-231 over nTNBC MCF-7 Cells.

Breast cancer (BC) is the first malignant neoplasm in women, with a high death rate despite early diagnoses and treatment advances. Significant differences exist between the most common BC and triple-negative breast cancer (TNBC). TNBC presents molecular differences such as lacking expression of the estrogen receptor (ER), progesterone receptor (PR), and HER2 proteins, making this cancer have a poor clinical prognostic and lack clear strategies for its treatment. However, growing evidence points to metabolic dysregulation as another differential process between stages and types of BC. Therefore, the study of this crucial hallmark could identify new therapeutic targets to treat this aggressive form of BC. These differences induce an in vitro exploration of the metabolic behavior of the MCF7 cells (nTNBC) and MDA-MB-231 (TNBC) cells under lipidomic based LC-MS. The results show more significant differences in lipid regulation (phosphatidylethanolamine) that could be associated with the aggressiveness and difficulties of the treatment of TNBC.

In Silico Drug Repositioning to Target the SARS-CoV-2 Main Protease as Covalent Inhibitors Employing a Combined Structure-Based Virtual Screening Strategy of Pharmacophore Models and Covalent Docking.

The epidemic caused by the SARS-CoV-2 coronavirus, which has spread rapidly throughout the world, requires urgent and effective treatments considering that the appearance of viral variants limits the efficacy of vaccines. The main protease of SARS-CoV-2 (Mpro) is a highly conserved cysteine proteinase, fundamental for the replication of the coronavirus and with a specific cleavage mechanism that positions it as an attractive therapeutic target for the proposal of irreversible inhibitors. A structure-based strategy combining 3D pharmacophoric modeling, virtual screening, and covalent docking was employed to identify the interactions required for molecular recognition, as well as the spatial orientation of the electrophilic warhead, of various drugs, to achieve a covalent interaction with Cys145 of Mpro. The virtual screening on the structure-based pharmacophoric map of the SARS-CoV-2 Mpro in complex with an inhibitor N3 (reference compound) provided high efficiency by identifying 53 drugs (FDA and DrugBank databases) with probabilities of covalent binding, including N3 (Michael acceptor) and others with a variety of electrophilic warheads. Adding the energy contributions of affinity for non-covalent and covalent docking, 16 promising drugs were obtained. Our findings suggest that the FDA-approved drugs Vaborbactam, Cimetidine, Ixazomib, Scopolamine, and Bicalutamide, as well as the other investigational peptide-like drugs (DB04234, DB03456, DB07224, DB7252, and CMX-2043) are potential covalent inhibitors of SARS-CoV-2 Mpro.

Untargeted LC-MS/MS Metabolomics Study on the MCF-7 Cell Line in the Presence of Valproic Acid.

To target breast cancer (BC), epigenetic modulation could be a promising therapy strategy due to its role in the genesis, growth, and metastases of BC. Valproic acid (VPA) is a well-known histone deacetylase inhibitor (HDACi), which due to its epigenetic focus needs to be studied in depth to understand the effects it might elicit in BC cells. The aim of this work is to contribute to exploring the complete pharmacological mechanism of VPA in killing cancer cells using MCF-7. LC-MS/MS metabolomics studies were applied to MCF-7 treated with VPA. The results show that VPA promote cell death by altering metabolic pathways principally pentose phosphate pathway (PPP) and 2'deoxy-α-D-ribose-1-phosphate degradation related with metabolites that decrease cell proliferation and cell growth, interfere with energy sources and enhance reactive oxygen species (ROS) levels. We even suggest that mechanisms such as ferropoptosis could be involved due to deregulation of L-cysteine. These results suggest that VPA has different pharmacological mechanisms in killing cancer cells including apoptotic and nonapoptotic mechanisms, and due to the broad impact that HDACis have in cells, metabolomic approaches are a great source of information to generate new insights for this type of molecule.

Targeting Several Biologically Reported Targets of Glioblastoma Multiforme by Assaying 2D and 3D Cultured Cells.

Glioblastoma multiforme (GBM) is account for 70% of all primary malignancies of the central nervous system. The median survival of human patients after treatment is around 15 months. There are several biological targets which have been reported that can be pursued using ligands with varied structures to treat this disease. In our group, we have developed several ligands that target a wide range of proteins involved in anticancer effects, such as histone deacetylase (HDACs), G protein-coupled estrogen receptor 1 (GPER), estrogen receptor-beta (ERß) and NADPH oxidase (NOX), that were screened on bidimensional (2D) and tridimensional (3D) GBM stem cells like (GSC). Our results show that some HDAC inhibitors show antiproliferative properties at 21-32 µM. These results suggest that in this 3D culture, HDACs could be the most relevant targets that are modulated to induce the antiproliferative effects that require in the future further experimental studies.

Myeloperoxidase Inhibitory and Antioxidant Activities of (E)-2-Hydroxy-α-aminocinnamic Acids Obtained through Microwave-Assisted Synthesis.

Myeloperoxidase (MPO) is an enzyme present in human neutrophils, whose main role is to provide defenses against invading pathogens. However, highly reactive oxygen species (ROS), such as HOCl, are generated from MPO activity, leading to chronic diseases. Herein, we report the microwave-assisted synthesis of a new series of stable (E)-(2-hydroxy)-α-aminocinnamic acids, in good yields, which are structurally analogous to the natural products (Z)-2-hydroxycinnamic acids. The radical scavenging activity (RSA), MPO inhibitory activity and cytotoxicity of the reported compounds were evaluated. The hydroxy derivatives showed the most potent RSA, reducing the presence of DPPH and ABTS radicals by 77% at 0.32 mM and 100% at 0.04 mM, respectively. Their mechanism of action was modeled with BDEOH, IP and ΔEH-L theoretical calculations at the B3LYP/6 - 31 + G(d,p) level. Compounds showed in vitro inhibitory activity of MPO with IC50 values comparable to indomethacin and 5-ASA, but cytotoxicities below 15% at 100-200 µM. Docking calculations revealed that they reach the amino acid residues present in the distal cavity of the MPO active site, where both the amino and carboxylic acid groups of the α-aminopropenoic acid arm are structural requirements for anchoring. (E)-2-hydroxy-α-aminocinnamic acids have been synthesized for the first time with a reliable method and their antioxidant properties demonstrated.

Learned Overweight Internal Model Can Be Activated to Maintain Equilibrium When Tactile Cues Are Uncertain: Evidence From Cortical and Behavioral Approaches.

Human adaptive behavior in sensorimotor control is aimed to increase the confidence in feedforward mechanisms when sensory afferents are uncertain. It is thought that these feedforward mechanisms rely on predictions from internal models. We investigate whether the brain uses an internal model of physical laws (gravitational and inertial forces) to help estimate body equilibrium when tactile inputs from the foot sole are depressed by carrying extra weight. As direct experimental evidence for such a model is limited, we used Judoka athletes thought to have built up internal models of external loads (i.e., opponent weight management) as compared with Non-Athlete participants and Dancers (highly skilled in balance control). Using electroencephalography, we first (experiment 1) tested the hypothesis that the influence of tactile inputs was amplified by descending cortical efferent signals. We compared the amplitude of P1N1 somatosensory cortical potential evoked by electrical stimulation of the foot sole in participants standing still with their eyes closed. We showed smaller P1N1 amplitudes in the Load compared to No Load conditions in both Non-Athletes and Dancers. This decrease neural response to tactile stimulation was associated with greater postural oscillations. By contrast in the Judoka's group, the neural early response to tactile stimulation was unregulated in the Load condition. This suggests that the brain can selectively increase the functional gain of sensory inputs, during challenging equilibrium tasks when tactile inputs were mechanically depressed by wearing a weighted vest. In Judokas, the activation of regions such as the right posterior inferior parietal cortex (PPC) as early as the P1N1 is likely the source of the neural responses being maintained similar in both Load and No Load conditions. An overweight internal model stored in the right PPC known to be involved in maintaining a coherent representation of one's body in space can optimize predictive mechanisms in situations with high balance constraints (Experiment 2). This hypothesis has been confirmed by showing that postural reaction evoked by a translation of the support surface on which participants were standing wearing extra-weight was improved in Judokas.

Modifications on the Tetrahydroquinoline Scaffold Targeting a Phenylalanine Cluster on GPER as Antiproliferative Compounds against Renal, Liver and Pancreatic Cancer Cells.

The implementation of chemo- and bioinformatics tools is a crucial step in the design of structure-based drugs, enabling the identification of more specific and effective molecules against cancer without side effects. In this study, three new compounds were designed and synthesized with suitable absorption, distribution, metabolism, excretion and toxicity (ADME-tox) properties and high affinity for the G protein-coupled estrogen receptor (GPER) binding site by in silico methods, which correlated with the growth inhibitory activity tested in a cluster of cancer cell lines. Docking and molecular dynamics (MD) simulations accompanied by a molecular mechanics/generalized Born surface area (MMGBSA) approach yielded the binding modes and energetic features of the proposed compounds on GPER. These in silico studies showed that the compounds reached the GPER binding site, establishing interactions with a phenylalanine cluster (F206, F208 and F278) required for GPER molecular recognition of its agonist and antagonist ligands. Finally, a 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) assay showed growth inhibitory activity of compounds 4, 5 and 7 in three different cancer cell lines-MIA Paca-2, RCC4-VA and Hep G2-at micromolar concentrations. These new molecules with specific chemical modifications of the GPER pharmacophore open up the possibility of generating new compounds capable of reaching the GPER binding site with potential growth inhibitory activities against nonconventional GPER cell models.

Somatosensory cortical facilitation during step preparation restored by an improved body representation in obese patients.

BACKGROUND: The anticipatory postural adjustments (APA) associated with step initiation are impaired in obese patients (e.g. longer duration, greater lateral center of pressure excursion). This could arise from the known altered internal representation of the body in obese individuals as this representation is crucial for enhancing the processing of foot cutaneous inputs prior to step initiation and for setting the APA. RESEARCH QUESTION: The purpose of the study was to examine if the processing of foot cutaneous inputs and the preparation of the APA when planning a step are impaired in obese patients due to their damaged body internal representation (BIR). We also investigated whether these sensorimotor processes will be restored after a 15-day intervention program composed of motor and cognitive activities engaging the BIR without aiming weight loss. METHODS: We compared, prior to (D1) and after (D15) the program, the amplitude of the cortical response evoked by foot cutaneous stimulation (SEP) occurring either during quiet standing or during the planning of a step in 18 obese patients (mean body mass index, BMI: 35). The APA were analyzed by measuring the amplitude and latency of the lateral force exerted on the ground. RESULTS AND SIGNIFICANCE: The SEP amplitude was not significantly different between the standing and stepping tasks at D1, but increased in the stepping task at D15. This enhanced sensory processing was associated with an increased activation of the posterior parietal cortex, suggesting a stronger involvement of the body representation during the planning of the stepping movement after the program. These cortical changes could have contributed to the changes in the temporal dimension of the APA observed at D15. These results suggest that programs targeting different dimensions of the BIR could be beneficial in improving the dynamic balance in obesity.

Percepción de riesgos asociados a estudios por imágenes en usuarios del Servicio de Radiología Oral de una Facultad de Odontología / Perception of risks associated with imaging studies in users of an Oral Radiology Service in a Dental School

Medir la percepción de riesgos asociados a los exámenes por imágenes y el nivel de conocimientos sobre los exámenes por imágenes, en usuarios del Servicio de Radiología Oral en una Facultad de Odontología de Lima. Material y Métodos: Se desarrolló un cuestionario para el estudio (V de Aiken: 0.97, Alfa de Cronbach: 0.70 para percepción de riesgo y 0.65 para nivel de conocimientos, y CCI: 0.81). El estudio incluyó 330 voluntarios entre 18 y 64 años de edad (68.8% mujeres, en su mayoría con un nivel de instrucción superior y nacidos en Lima. La mayor proporción de puntajes para percepción de riesgo asociado a estudios por imágenes y para el nivel de conocimientos sobre exámenes de imágenes, fueron de nivel intermedio. Resultados: El sexo femenino estuvo asociado con un mayor nivel de percepción de riesgo (X2 p=0,007), así como las edades más altas estuvieron asociadas a un mayor nivel de conocimientos (X2 p=0.023). Conclusiones: Los usuarios del Servicio de Radiología Oral de la Clínica Estomatológica Central de la Facultad de Estomatología de la Universidad Peruana Cayetano Heredia en el año 2013, fueron en su mayor parte mujeres, mayores de 45 años de edad, con un nivel de instrucción superior, nacidos en la ciudad de Lima y procedentes de distritos vecinos a la institución. Los puntajes de la percepción sobre riesgos asociados a los estudios por imágenes fueron en mayor proporción del nivel intermedio. Los puntajes del nivel de conocimientos sobre los riesgos, fueron también en mayor proporción del nivel intermedio. El sexo femenino estuvo asociado con el mayor nivel de percepción de riesgo, y lo mismo que la mayor edad estuvo asociada a un mayor nivel de conocimientos...

Objective: Measure the perception of risks associated with imaging tests and the level of knowledge on imaging tests on users of the Oral Radiology School of Dentistry in Lima. Methods: A questionnaire for the study was developed (V Aiken: 0.97, CronbachÆs alpha: 0.70 for Risk Perception and 0.65 for level of knowledge, and CCI: 0.81). The study included 330 volunteers aged between 18 and 64 years (68.8% women), aged 45 years, mostly with a higher level of instruction and born in Lima. The largest proportion of scores for perception of risk associated with imaging and to the level of knowledge on imaging tests were intermediate level. Results: Female sex was associated with a higher level of risk perception (X2 p=0,007) and higher ages were associated with a higher level of knowledge (X2 p=0.023). Conclusions: The users of the Oral Radiology Department of the Central Clinic of the Dentistry School at the Universidad Peruana Cayetano Heredia in 2013, were mostly women over 45 years old, with a higher level of instruction, born in Lima and from neighboring districts to the institution. Scores of Perception about risks associated with imaging studies were greater proportion of intermediate level. Scores on the level of knowledge about the risks were also higher proportion of intermediate level. Female sex is associated with higher levels of perceived risk, and so that older age is associated with a higher level of knowledge...

Relación entre la edad cronológica y la mineralización del tercer molar inferior según método de Demirjian / Relationship between chronological age and mineralization of the lower third molar according to Demirjian stages

Objetivos: Relacionar la edad cronológica y la mineralización del tercer molar inferior según los estadios de Demirjian en radiografías panorámicas digitales. Material y Métodos: Se realizó un análisis retrospectivo evaluando 1176 radiografías panorámicas digitales del Servicio de Radiología de la Clínica Dental de la Facultad de Estomatología, Universidad Peruana Cayetano Heredia, entre los años 2011 y 2012. Los casos tenían entre 7 - 23 años de edad. Resultados: En la pieza 38, el estadio D presentó una edad promedio de 14,05 ± 1,35 años para el sexo femenino y 13,42 ± 1,30 años para el sexo masculino. El tercer molar inferior en el sexo masculino presentó una maduración ligeramente anterior que el sexo femenino, esta diferencia fue estadísticamente significativa en los estadios D, E, F, G y H. No se encontró diferencia entre los estadios de Demirjian de la pieza 38 y 48. En el estadio H el 100% del sexo femenino y el 99,1% del sexo masculino tuvieron 18 o más años. Conclusiones: Los resultados sugieren que el método de Demirjian, para evaluar la mineralización del tercer molar, puede ser una herramienta útil en el estudio de la edad cronológica.

Objectives: To relate the chronological age and the mineralization of the lower third molar according to Demirjian stages on digital panoramic radiographs. Methods: A retrospective analysis was conducted, evaluating 1176 digital panoramic radiographs from the Dental Clinic radiology area of the School of Dentistry at Universidad Peruana Cayetano Heredia, between 2011 and 2012. The cases were aged 7-23 years. Results: In tooth 38, the stage D represented an average age of 14.05 ± 1.35 years for females and 13.42 ± 1.30 years for males. The third molar presented a slightly earlier maturation in males than in females, this difference was statistically signifi cant in stages D, E, F, G and H. No difference was found between the stages of Demirjian in teeth 38 and 48. In stage H 100% female and 99.1% male had 18 or more years. Conclusions: The results suggest that using Demirjian stages to assess the mineralization of third molar can be a useful tool in chronological age study.

Synthesis of pyrrolidinoindolines from 2-(2-oxo-3-indolyl)acetates: scope and limitations.

A series of 1,3a,8-alkylpyrrolidinoindolines have been synthesized. The scope and limitations of the alkylation of starting methyl oxindol-3-acetates are explored employing electron-rich and electron-poor alkylating agents. Hydrolysis and reductive lactonization of the resulting carboxylic gamma-oxindolic acid derivatives proceeds with good yields to afford 2-oxofuroindolines providing ready access to the pyrrolidinoindoline derivatives.