Factores clínicos asociados a complicaciones pulmonares y extrapulmonares en pacientes con fibrosis quística. Un estudio observacional de centro único / Clinical factors associated with pulmonary and extrapulmonary complications in patients with cystic fibrosis. A single-center observational study

Introducción: La fibrosis quística (FQ), afecta el epitelio exocrino, formando una mucosidad espesa que obstruye los conductos de los diferentes órganos, siendo el pulmón y páncreas los más afectados. El objetivo del presente estudio fue determinar la asociación entre los factores clínicos y las complicaciones, así como la supervivencia de pacientes con FQ atendidos en un centro de referencia pediátrica en México. Métodos: El presente estudio observacional, longitudinal, fue realizado en el Instituto Nacional de Pediatría en México, de abril del 2012, a abril del 2022. Con una muestra no probabilística ingresaron al estudio niños con fibrosis quística. Las variables fueron: demográficas, complicaciones pulmonares, extrapulmonares y mortalidad. Se realiza la asociación con chi-cuadrado y la supervivencia con Kaplan-Meir. Resultados: Se analizan 71 pacientes, lactantes menores 41 casos (62%), pre-escolares 12 casos (16.9%). Fueron 40 hom-bres (56.3%). 57 casos (80.3%) tuvieron exacerbación infecciosa, 11 casos (15.5%) aspergi-losis broncopulmonar, 9 casos (12.7%) con hipertensión pulmonar, 5 casos (7%) con neumotórax, 12 casos (16.9%) con ventilación mecánica, 70 casos (98.5) con insuficiencia pancreática, 10 casos (14%) con enfermedad hepatobiliar. Hubo asociación de complica-ciones extrapulmonares con el fenotipo F508. La supervivencia global fue de 150 meses. En pacientes con ventilación mecánica invasiva la supervivencia fue de 55 meses (P<0.001), en pacientes con ventilación mecánica no invasiva fue de 106 meses (P<0.001), en pacientes con neumotórax fue de 25 meses P<0.001, en pacientes con aspergilosis bronco-pulmonar alérgica fue de 125 meses P<0.01. Conclusión: la mayor compli-cación extrapulmonar de pa-cientes con FQ es la insuficien-cia pancreática. La presencia de neumotórax disminuye la supervivencia.

Introduction: Cystic fibrosis (CF) affects the exocrine epithelium, forming a thick mucus that obstructs the ducts of the different organs, with the lungs and pancreas being the most affected. This study aimed to determine the association between clinical factors and complications, as well as the survival of CF patients treated at a pediatric reference center in Mexico. Methods: This observational, longitudinal study was conducted at the National Institute of Pediatrics in Mexico from April 2012 to April 2022. Children with cystic fibrosis were entered into the study with a nonprobabilistic sample. The variables were demographic, pulmonary and extrapulmonary complications and mortality. The association was made with chi-square and survival with Kaplan- Meir. Results: Seventy-one patients were analyzed, including 41 infants (62%) and 12 preschoolers (16.9%). There were 40 men (56.3%). Fifty-seven cases (80.3%) had an infectious exacerbation, 11 cases (15.5%) had allergic bronchopulmonary aspergillosis, 9 cases (12.7%) had pulmonary hypertension, 5 cases (7%) had pneumothorax, 12 cases (16.9%) had mechanical ventilation, 70 cases (98.5%) had pancreatic insufficiency, and 10 cases (14%) had hepatobiliary disease. There was an association of extrapulmonary complications with the F508 phenotype. Overall survival was 150 months. In patients with invasive mechanical ventilation, survival was 55 months (P<0.001); in patients with noninvasive mechanical ventilation, it was 106 months (P<0.001); in patients with pneumothorax, it was 25 months (P<0.001); and in patients with allergic bronchopulmonary aspergillosis, it was 125 months (P<0.01). Conclusion: The most significant extrapulmonary complication in CF patients is pancreatic insufficiency. The presence of pneumothorax markedly decreases survival.

Resting Energy Expenditure Prediction Equations in the Pediatric Population: A Systematic Review.

Background and Aims: The determination of energy requirements is necessary to promote adequate growth and nutritional status in pediatric populations. Currently, several predictive equations have been designed and modified to estimate energy expenditure at rest. Our objectives were (1) to identify the equations designed for energy expenditure prediction and (2) to identify the anthropometric and demographic variables used in the design of the equations for pediatric patients who are healthy and have illness. Methods: A systematic search in the Medline/PubMed, EMBASE and LILACS databases for observational studies published up to January 2021 that reported the design of predictive equations to estimate basal or resting energy expenditure in pediatric populations was carried out. Studies were excluded if the study population included athletes, adult patients, or any patients taking medications that altered energy expenditure. Risk of bias was assessed using the Quality Assessment Tool for Observational Cohort and Cross-Sectional Studies. Results: Of the 769 studies identified in the search, 39 met the inclusion criteria and were analyzed. Predictive equations were established for three pediatric populations: those who were healthy (n = 8), those who had overweight or obesity (n = 17), and those with a specific clinical situation (n = 14). In the healthy pediatric population, the FAO/WHO and Schofield equations had the highest R 2 values, while in the population with obesity, the Molnár and Dietz equations had the highest R 2 values for both boys and girls. Conclusions: Many different predictive equations for energy expenditure in pediatric patients have been published. This review is a compendium of most of these equations; this information will enable clinicians to critically evaluate their use in clinical practice. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=226270, PROSPERO [CRD42021226270].

Evaluación de la asimetría mamaria mediante algoritmos de desplazamiento óptico / Breast asymmetry assessment using optical flow algorithms

Antecedentes y Objetivo. La gradación de la simetría es un aspecto fundamental para evaluar un resultado en cirugía mamaria. Los métodos más extendidos de valoración de asimetría se basan en percepciones subjetivas del cirujano o de la paciente, así como en la comparación objetiva de distancias entre puntos de interés tales como la horquilla esternal y el pezón. Utilizar de forma generalizada un método objetivo comportaría ventajas evidentes. El presente trabajo pretende diseñar y contrastar un método de análisis de imagen que permita objetivar el grado de asimetría mamaria en una paciente. Material y método. Basándonos en un algoritmo informático que evalúa el grado de desplazamiento entre dos imágenes similares, desarrollamos una aplicación capaz de cuantificar el grado de desplazamiento que presenta una mama con respecto a la otra, arrojando un resultado numérico que denominamos global breast asymmetry index (GBAI). Calculamos este valor en una serie de 50 pacientes de forma pre y postoperatoria. Comparamos el resultado con otros métodos de evaluación de asimetría tanto objetivos como subjetivos, incluyendo una encuesta a 100 profesionales sanitarios y a un panel de expertos. Resultados. La adaptación de nuestro algoritmo para calcular un índice objetivo de asimetría mamaria muestra un comportamiento similar a la valoración subjetiva de la asimetría por parte del personal sanitario, mejorando otros métodos objetivos y presentándose como una alternativa útil a los métodos tradicionales de comparación de distancias. Conclusiones. El cálculo de desplazamiento óptico es un procedimiento válido y objetivo para cuantificar la asimetría mamaria

Background and Objective. Adequate assessment of symmetry is a critical aspect when evaluating results in breast surgery. Most methods of asymmetry assessment are based on subjective observations by the surgeon or patients, as well as on the objective measurement of distances between key points, such as the sternal notch and the nipple. The widespread use of an objective method would have obvious advantages. We aim to design and test an imaging analysis method that allows to objectively assess the degree of breast asymmetry. Methods. An application has been developed to quantify the degree of disparity between two similar images, based on a computer algorithm. The algorithm was used to compare images of breasts, yielding a numerical result that we named global breast asymmetry index (GBAI). This value was calculated in a series of 50 patients pre- and postoperatively. The result was compared with other asymmetry assessment methods, both objective and subjective, including a survey of 100 health professionals and a panel of experts. Results. The implementation of our algorithm to calculate the breast asymmetry index shows similar results to subjective assessment of asymmetry by health personnel, improving other objective methods and presenting itself as a useful alternative to traditional methods of symmetry assessment. Conclusions. Measurement of optical flow is a valid and objective method of assessment of the degree of breast asymmetry

IKZF1 Gene Deletion in Pediatric Patients Diagnosed with Acute Lymphoblastic Leukemia in Mexico.

The IKZF1 gene is formed by 8 exons and encodes IKAROS, a transcription factor that regulates the expression of genes that control cell cycle progression and cell survival. In general, 15-20% of the patients with preB acute lymphoblastic leukemia (preB ALL) harbor IKZF1 deletions, and the frequency of these deletions increases in BCR-ABL1 or Ph-like subgroups. These deletions have been associated with poor treatment response and the risk of relapse. The aim of this descriptive study was to determine the frequency of IKZF1 deletions and the success of an induction therapy response in Mexican pediatric patients diagnosed with preB ALL in 2 hospitals from 2017 to August 2018. Thirty-six bone marrow samples from patients at the Instituto Nacional de Pediatría in Mexico City and the Centro Estatal de Cancerología in Tepic were analyzed. The IKZF1 deletion was identified by MLPA using the SALSA MLPA P335 ALL-IKZF1 probemix. Deletions of at least 1 IKZF1 exon were observed in 7/34 samples (20.6%): 3 with 1 exon deleted; 1 with 2 exons, 1 with 5 exons, 1 with 6 exons, and 1 patient with a complete IKZF1 deletion. This study was descriptive in nature; we calculated the frequency of the IKZF1 gene deletion in a Mexican pediatric population with preB ALL as 20.6%.

Impact of Regional Anesthesia on Gastroesophageal Cancer Surgery Outcomes: A Systematic Review of the Literature.

Regional anesthesia may play a beneficial role in long-term oncological outcomes. Specifically, it has been suggested that it can prolong recurrence-free survival and overall survival after gastrointestinal cancer surgery, including gastric and esophageal cancer, by modulating the immune and inflammatory response. However, the results from human studies are conflicting. The goal of this systematic review was to summarize the evidence on the impact of regional anesthesia on immunomodulation and cancer recurrence after gastric and esophageal surgery. We conducted a literature search of 5 different databases. Two independent reviewers analyzed the quality of the selected manuscripts according to prespecified inclusion and exclusion criteria. Randomized controlled trials were assessed for potential sources of bias by using the Cochrane Risk of Bias tool. A total of 6 studies were included in the quality analysis and systematic review. A meta-analysis was not conducted for several reasons, including high heterogeneity among studies, low quality of the reports, and lack of standardized outcomes definitions. Although the literature suggests that regional anesthesia has some modulatory effects on the inflammatory and immunological response in the studied patient population, our systematic review indicates that there is no evidence to support or refute the use of epidural anesthesia or analgesia with the goal of reducing cancer recurrence after gastroesophageal cancer surgery.

Krüppel Like Factors Family Expression in Cervical Cancer Cells.

BACKGROUND AND AIMS: Krüppel Like Factors (KLF) refers to a family of seventeen members of transcription factors. Involved in several cellular processes. As other cancer types, Cervical Cancer (CC) presents molecular deregulations in transcription factors, but especially Human Papilloma Virus (HPV) sequences. Here in this work we analyzed the mRNA expression of all KLF family members in CC-derived cell lines and CC tissues. METHODS: The cell lines used were HeLa, INBL, RoVa, C4-I, Ms751, ViPa, CaLo, SiHa, CaSki, C33a and ViBo and the non-tumorigenic HaCaT. mRNA expression was analyzed by means of expression microarray and RT-PCR, and KLF5 protein by immunofluorescence. RESULTS: The cell lines were grouped according to HPV genotype as HPV16, HPV18 positive or HPV negative cells. Heterogeneous expression was observed among the cell lines. Despite the heterogeneous expression profile, KLF3, -5, -12, -15 and -16 transcripts were present in all cell lines, KLF4 and -10 which were not expressed in CaSki; KLF11 and 13 were not expressed by Vipa and C4-I, and KLF7 was not expressed by C4-I and Rova. The CC tissue analysis shows expression of most of the KLF members, such as KLF5. KLF5 immunosignal was positive in the three cell lines analyzed. CONCLUSIONS: We suggest that KLF expression could not be related to HPV presence/genotype, at least at transcriptional level, and the expression of KLF family members may be necessary in the biology of the CC cells.

Impact of Regional Anesthesia on Recurrence, Metastasis, and Immune Response in Breast Cancer Surgery: A Systematic Review of the Literature.

BACKGROUND AND OBJECTIVES: The perioperative period is critical in the long-term prognosis of breast cancer patients. The use of regional anesthesia, such as paravertebral block (PVB), could be associated with improvements in long-term survival after breast cancer surgery by modulating the inflammatory and immune response associated with the surgical trauma, reducing opioid and general anesthetic consumption, and promoting cancer cells death by a direct effect of local anesthetics. METHODS: A systematic literature search was conducted for studies of patients who received PVB for breast cancer surgery. The Jadad score and Ottawa-Newcastle scale were used to assess the methodological quality of randomized controlled trial and observational retrospective studies, respectively. Only high-quality studies were considered for meta-analysis. The selected studies were divided into 3 groups to determine the impact of PVB on (a) recurrence and survival, (b) humoral response, and (c) cellular immune response. RESULTS: We identified 467 relevant studies; 121 of them underwent title and abstract review, 107 were excluded, and 15 studies were selected for full text reading and quality assessment. A meta-analysis was not conducted because of low-quality studies and lack of uniform definition among primary outcomes. Thus, a systematic review of the current evidence was performed. CONCLUSIONS: Our study indicates that there are no data to support or refute the use of PVB for reduction of cancer recurrence or improvement in cancer-related survival. However, PVB use is associated with lower levels of inflammation and a better immune response in comparison with general anesthesia and opioid-based analgesia.

N-(2-hydroxyphenyl)-2-propylpentanamide, a valproic acid aryl derivative designed in silico with improved anti-proliferative activity in HeLa, rhabdomyosarcoma and breast cancer cells.

Epigenetic alterations are associated with cancer and their targeting is a promising approach for treatment of this disease. Among current epigenetic drugs, histone deacetylase (HDAC) inhibitors induce changes in gene expression that can lead to cell death in tumors. Valproic acid (VPA) is a HDAC inhibitor that has antitumor activity at mM range. However, it is known that VPA is a hepatotoxic drug. Therefore, the aim of this study was to design a set of VPA derivatives adding the arylamine core of the suberoylanilide hydroxamic acid (SAHA) with different substituents at its carboxyl group. These derivatives were submitted to docking simulations to select the most promising compound. The compound 2 (N-(2-hydroxyphenyl)-2-propylpentanamide) was the best candidate to be synthesized and evaluated in vitro as an anti-cancer agent against HeLa, rhabdomyosarcoma and breast cancer cell lines. Compound 2 showed a better IC50 (µM range) than VPA (mM range) on these cancer cells. And also, 2 was particularly effective on triple negative breast cancer cells. In conclusion, 2 is an example of drugs designed in silico that show biological properties against human cancer difficult to treat as triple negative breast cancer.

Identification of the antiepileptic racetam binding site in the synaptic vesicle protein 2A by molecular dynamics and docking simulations.

Synaptic vesicle protein 2A (SV2A) is an integral membrane protein necessary for the proper function of the central nervous system and is associated to the physiopathology of epilepsy. SV2A is the molecular target of the anti-epileptic drug levetiracetam and its racetam analogs. The racetam binding site in SV2A and the non-covalent interactions between racetams and SV2A are currently unknown; therefore, an in silico study was performed to explore these issues. Since SV2A has not been structurally characterized with X-ray crystallography or nuclear magnetic resonance, a three-dimensional (3D) model was built. The model was refined by performing a molecular dynamics simulation (MDS) and the interactions of SV2A with the racetams were determined by docking studies. A reliable 3D model of SV2A was obtained; it reached structural equilibrium during the last 15 ns of the MDS (50 ns) with remaining structural motions in the N-terminus and long cytoplasmic loop. The docking studies revealed that hydrophobic interactions and hydrogen bonds participate importantly in ligand recognition within the binding site. Residues T456, S665, W666, D670 and L689 were important for racetam binding within the trans-membrane hydrophilic core of SV2A. Identifying the racetam binding site within SV2A should facilitate the synthesis of suitable radio-ligands to study treatment response and possibly epilepsy progression.

Heterogeneity of microRNAs expression in cervical cancer cells: over-expression of miR-196a.

In recent years, the study of microRNAs associated with neoplastic processes has increased. Patterns of microRNA expression in different cell lines and different kinds of tumors have been identified; however, little is known about the alterations in regulatory pathways and genes involved in aberrant set of microRNAs. The identification of these altered microRNAs in several cervical cancer cells and potentially deregulated pathways involved constitute the principal goals of the present study. In the present work, the expression profiles of cellular microRNAs in Cervical Cancer tissues and cell lines were explored using microRNA microarray, Affymetrix. The most over-expressed was miR-196a, which was evaluated by real time PCR, and HOXC8 protein as potential target by immunohistochemistry assay. One hundred and twenty three human microRNAs differentially expressed in the cell tumor, 64 (52%) over-expressed and 59 (48%) under-expressed were observed. Among the microRNAs over-expressed, we focused on miR-196a; at present this microRNA is poorly studied in CC. The expression of this microRNA was evaluated by qRT-PCR, and HOXC8 by immunohistochemistry assay. There is not a specific microRNA expression profile in the CC cells, neither a microRNA related to HPV presence. Furthermore, the miR-196a was over-expressed, while an absence of HOXC8 expression was observed. We suggest that miR-196a could be played as oncomiR in CC.

Splice variants of zinc finger protein 695 mRNA associated to ovarian cancer.

BACKGROUND: Studies of alternative mRNA splicing (AS) in health and disease have yet to yield the complete picture of protein diversity and its role in physiology and pathology. Some forms of cancer appear to be associated to certain alternative mRNA splice variants, but their role in the cancer development and outcome is unclear. METHODS: We examined AS profiles by means of whole genome exon expression microarrays (Affymetrix GeneChip 1.0) in ovarian tumors and ovarian cancer-derived cell lines, compared to healthy ovarian tissue. Alternatively spliced genes expressed predominantly in ovarian tumors and cell lines were confirmed by RT-PCR. RESULTS: Among several significantly overexpressed AS genes in malignant ovarian tumors and ovarian cancer cell lines, the most significant one was that of the zinc finger protein ZNF695, with two previously unknown mRNA splice variants identified in ovarian tumors and cell lines. The identity of ZNF695 AS variants was confirmed by cloning and sequencing of the amplicons obtained from ovarian cancer tissue and cell lines. CONCLUSIONS: Alternative ZNF695 mRNA splicing could be a marker of ovarian cancer with possible implications on its pathogenesis.

Exploration of the valproic acid binding site on histone deacetylase 8 using docking and molecular dynamic simulations.

Epigenetic therapy is an important focus of research for drug development in the treatment of cancer. Valproic acid (VPA) is an HDAC inhibitor that has been evaluated in clinical studies. Despite its success in treating cancer, the mechanism of inhibition of VPA in HDAC is unknown. To this end, we have used docking and molecular dynamic simulations to investigate VPA binding to HDAC, employing both native and rebuilt 3-D structures. The results showed that VPA, via its carboxyl group, coordinates the Zn atom and other local residues (H141-142 and Y360) located at the catalytic site (CS) of HDAC. This causes electrostatic and hydrogen bonding interactions while having little interaction with the hydrophobic side chains, resulting in a low affinity. However, after several docking studies on different native HDAC 3-D structures and after using several snapshots from MD simulations, it became apparent that VPA bound with highest affinity at a site located at the acetyl-releasing channel, termed the hydrophobic active site channel (HASC). The affinity of VPA for HASC was due to its highly hydrophobic properties that allow VPA to take part in van der Waals interactions with Y18, I19, Y20, V25, R37, A38, V41, H42, I135 and W137, while VPA's carboxylate group has several hydrogen bonding interactions with the backbones of S138, I19, N136 and W137. MD simulations showed that the HASC door continuously opened and closed, which affected the affinity of VPA to the HASC, but the affinity toward the HASC was consistently higher than that obtained for the CS, suggesting that the HASC could be involved in the mechanism of inhibition.

Early death in children with acute lymphoblastic leukemia: does malnutrition play a role?

The study aim was to correlate malnutrition and early death in children with acute lymphoblastic leukemia (ALL). A study was conducted in 100 consecutive children with ALL. An analysis included clinical and laboratory parameters as well as co-morbidity factors. Forty patients were standard risk and 60 high risk. Multivariate analysis showed variables of statistical importance, including female gender (p 010), ALL high-risk (p 04), and infection (p 036). Malnutrition (p 1.0) and poverty (p 0.5) did not influence. Early mortality was documented in 15/100 (15%) patients. The study shows that high-risk ALL and infection represent the leading causes of early mortality.

Prognostic factors including neoadjuvant chemotherapy in Mexican children with neuroblastoma.

INTRODUCTION: There are several prognostic factors in children with neuroblastoma that have been outlined in the international literature. MATERIAL AND METHODS: A retrospective study was carried out analysing the medical records of patients with the pathological diagnosis of neuroblastoma seen at the Department of Oncology from the Instituto Nacional de Pediatriá (Mexico) between January 1984 to January 1997. A total of 32 clinical prognostic factors were assess in our population. RESULTS: Fifty five patients whose age ranged from 1 to 168 months old, mean of 35 months were included. Out of 32 prognostic factors only 6 including sex (p= 0.0039), metastatic disease to bone (p= 0.003), bone marrow involvement (p= 0.0027), staging system (p= 0.000015), surgical treatment (p 0,0022) and neoadjuvant chemotherapy (p.005) were the most significant. CONCLUSIONS: It was concluded that besides the prognostic factors outlined, neoadjuvant chemotherapy is of utmost importance. It decreases tumor volume and allows surgery to be more successful, therefore believing that this variable represents a specific prognostic factor in cases of advanced neuroblastoma.

Prognostic factors including neoadjuvant chemotherapy in Mexican children with neuroblastoma

Introducción. Existen diferentes factores pronósticos en los niños con neuroblastoma, estos factores tienen diferente peso específico de acuerdo a la literatura internacional. Material y métodos. Se realizó un estudio retrospectivo, analizando los expedientes clínicos de los pacientes con el diagnóstico de neuroblastoma, tratados en el Instituto Nacional de Pediatría (México) de enero de 1984 a enero de 1997. Se analizaron un total de 32 factores pronósticos clínicos dentro de nuestra población. Resultados. Se incluyeron un total de 55 pacientes, con un rango de edad de 1 a 68 meses, con una media de 36 meses de edad. De los 32 factores pronósticos evaluados, únicamente 6 fueron significativos, los cuales incluyeron el sexo (p= 0,0039), enfermedad metastásica a hueso (p= 0,003), infiltración a médula ósea (p= 0,0027), estadificación (p= 0,000015), tratamiento quirúrgico (p= 0,0022) y el uso de quimioterapia neoadyuvante (p= 0,005). Conclusiones. En análisis de los resultados de los diferentes factores pronósticos nos permiten concluir que la quimioterapia neoadyuvante realiza un papel importante al permitir la disminución del volumen tumoral, lo que permite que la resección quirúrgica pueda ser completa. El resto de las variables que fueron significativas están asociadas directamente con la presencia de neuroblastoma en estadio avanzado

Introduction. There are several prognostic factors in children with neuroblastoma that have been outlined in the international literature. Material and methods. A retrospective study was carried out analysing the medical records of patients with the pathological diagnosis of neuroblastoma seen at the Department of Oncology from the Instituto Nacional de Pediatría (Mexico) between January 1984 to January 1997. A total of 32 clinical prognostic factors were assess in our population. Results. Fifty five patients whose age ranged from 1 to 168 months old, mean of 35 months were included. Out of 32 prognostic factors only 6 including sex (p= 0.0039), metastatic disease to bone (p= 0.003), bone marrow involvement (p= 0.0027), staging system (p= 0.000015), surgical treatment (p 0,0022) and neoadjuvant chemotherapy (p.005) were the most significant. Conclusions. It was concluded that besides the prognostic factors outlined, neoadjuvant chemotherapy is of utmost importance. It decreases tumor volume and allows surgery to be more successful, therefore believing that this variable represents a specific prognostic factor in cases of advanced neuroblastoma

Grado de penetración de bebidas de uso común a la estructura dental / Degree of leakage of common beverages on the dental structure

El propósito del trabajo fue determinar la permeabilidad del esmalte, la dentina y el cemento a bebidas de uso corriente (café, vino tinto y cola) y evaluar las consecuencias estéticas que pudiera tener. El estudio fue "in vitro", con dientes recién extraídos, despulpados e hidratados, con la cavidad pulpar sellada. Catorce dientes se expusieron a cola (Coca Cola), 12a café y 12a vino tinto. Transcurridos 31 días se realizó un corte longitudinal a cada diente para observar y medir, por medio de un microscopio metalográfico, la penetración de cada bebida a la estructura dentaria.- La cola fue la que más profundamente penetró en el esmalte (106.771l), en el que produjo una pigmentación de color pardo oscuro; la bebida de menor penetración fue el café (23.651l) y produjo una coloración amarillo clara. En la raíz (cemento y dentina) la mayor penetración fue para el vino tinto (promedio de 123.98 u).