Importance of the vaporization margin during CO2 laser treatment of oral leukoplakia: A survival study.

OBJECTIVES: The main purpose of this study was to assess the response of oral leukoplakia to CO2 laser vaporization treatment, as well as determining possible factors that may affect recurrence of lesions. MATERIALS AND METHODS: A retrospective study was conducted, in which the medical records of patients who had been clinically and histologically diagnosed with oral leukoplakia and treated with CO2 laser between 1996 and 2019 at the Oral Medicine Teaching Unit of the Faculty of Dentistry of the University of Santiago de Compostela were reviewed. RESULTS: Fifty-eight patients were included: 36 female and 22 male subjects, with a mean age of 63.7 years old (SD ±13.1). The average follow-up time was 57.5 months (SD ±57.9). A relapse rate of 52.6% was determined. Of all the studied variables, the margin was the only one for which a statistically significant association with recurrence of lesions was demonstrated (p < 0.05). CONCLUSION: The vaporization of lesions using CO2 laser with a safety margin of at least 3 mm from the clinical limits of OL is a key factor in preventing recurrence.

Circulating tumor DNA tracking in patients with pancreatic cancer using next-generation sequencing.

BACKGROUND: Pancreatic cancer remains one of the most devastating malignancies due to the absence of techniques for early diagnosis and the lack of target therapeutic options for advanced disease. Next Generation Sequencing (NGS) generates high throughput and valuable genetic information when evaluating circulating tumor DNA (ctDNA); however clinical utility of liquid biopsy in pancreatic cancer has not been demonstrated yet. The aim of this study was to evaluate whether results from a Next Generation Sequencing panel on plasma samples from pancreatic cancer patients could have a clinical significance. METHODS: From December 2016 to January 2020, plasma samples from 27 patients with pancreatic ductal adenocarcinoma at two different tertiary Spanish Hospitals underwent ctDNA testing using a commercial NGS panel of 65 genes. Clinical data were available for these patients. VarsSome Clinical software was used to analyse NGS data and establish pathogenicity. RESULTS: Evaluable NGS results were obtained in 18 out of the 27 plasma samples. Somatic pathogenic mutations were found mainly in KRAS, BRCA2, FLT3 and HNF1A, genes. Pathogenic mutations were detected in 50% of plasma samples from patient diagnosed at stages III-IV samples. FLT3 mutations were observed in 22.22% of samples which constitute a novel result in the field. CONCLUSIONS: Liquid biopsy using NGS is a valuable tool but still not sensitive or specific enough to provide clinical utility in pancreatic cancer patients.

Supervised Machine Learning Methods and Hyperspectral Imaging Techniques Jointly Applied for Brain Cancer Classification.

Hyperspectral imaging techniques (HSI) do not require contact with patients and are non-ionizing as well as non-invasive. As a consequence, they have been extensively applied in the medical field. HSI is being combined with machine learning (ML) processes to obtain models to assist in diagnosis. In particular, the combination of these techniques has proven to be a reliable aid in the differentiation of healthy and tumor tissue during brain tumor surgery. ML algorithms such as support vector machine (SVM), random forest (RF) and convolutional neural networks (CNN) are used to make predictions and provide in-vivo visualizations that may assist neurosurgeons in being more precise, hence reducing damages to healthy tissue. In this work, thirteen in-vivo hyperspectral images from twelve different patients with high-grade gliomas (grade III and IV) have been selected to train SVM, RF and CNN classifiers. Five different classes have been defined during the experiments: healthy tissue, tumor, venous blood vessel, arterial blood vessel and dura mater. Overall accuracy (OACC) results vary from 60% to 95% depending on the training conditions. Finally, as far as the contribution of each band to the OACC is concerned, the results obtained in this work are 3.81 times greater than those reported in the literature.

CA19-9 capability as predictor of pancreatic cancer resectability in a Spanish cohort.

CA19-9 serum has been suggested as a marker of unresectability but different cut-off levels have been published. A cut-off of 500 U/ml is currently considered in an international consensus as biological criteria of borderline resectable pancreatic adenocarcinoma. To evaluate whether serum CA19-9 threshold of 500 U/ml could be adequate predictor of resectability in pancreatic adenocarcinoma. Multicenter, observational, prospective study performed in Spain including 203 patients diagnosed with pancreatic adenocarcinoma. 43 (21.2%) cases were resectable and 160 (78.8%) unresectable. Among the 176 preoperative CA19-9 available values, 98 (58.3%) were ≤ 500 U/ml and 73 (42.7%) > 500 U/ml. Resectability rate in those patients with CA19-9 ≤ 500 U/ml was 60% while it was found to be 18% when CA19-9 > 500 U/ml. Statistical model to predict resectability based on CA19-9 provide an AUC of 0.6618 (95% CI 0.53-0.83) when only CA19-9 values > 500 U/ml are studied. Serum levels of CA19-9 higher than 500 U/ml are indicative of unresectable disease, however reduced sensitivity and specificity lead to a limited clinical applicability for resectability.

Psychological disorders and quality of life in oral lichen planus patients and a control group.

OBJECTIVES: Oral lichen planus (OLP) is a common disease whose aetiopathogenesis is linked to psychological disorders. This study aims to determine the influence of anxiety and depression in OLP patients, define the perception of quality of life in these patients and check for potential differences between atrophic/ ulcerative and reticular lesions. MATERIALS AND METHODS: A group of 48 OLP patients and a control group of 40 patients were selected. In order to assess anxiety, depression and quality of life, Hospital Anxiety and Depression Scale (HADS) and Oral Health Impact Profile 14 (OHIP-14) tests were completed. RESULTS: OLP patients showed higher scores on HADS (anxiety p < 0.01, depression p < 0.05) and OHIP-14 (physical pain p < 0.05, psychological discomfort p = 0.001). Patients with reticular lesions obtained higher scores in HADS (anxiety p = 0.001, depression p < 0.001), whereas patients with atrophic/ ulcerative lesions obtained higher scores in OHIP-14 (p = 0.02). CONCLUSIONS: Psychological disorders play an important role as a trigger for OLP and are responsible for many relapses. Psychological support would be advisable in order to improve their mental health, as this would have a positive impact on their quality of life and would lead to a better progression of the disease.

Removal of Two Neonicotinoid Insecticides and Mineralization of 14C-Imidacloprid in Biomixtures.

Environmental contamination with neonicotinoid insecticides represents an issue of wide concern due to their negative effects on pollinators. The goal of this work was to evaluate the potential use of biomixtures employed in biopurification systems (BPS) to remove two neonicotinoid pesticides, imidacloprid and thiamethoxam, from wastewater of agricultural origin. The removal was assayed by quantification of the parent compounds and the detection of putative transformation products of imidacloprid by means of LC-MS/MS, and mineralization of radiolabeled imidacloprid. Two biomixtures (B1, B2) were prepared using coconut fiber, compost and two soils pre-exposed to imidacloprid (volumetric composition 50:25:25). After spiking of neonicotinoids and 228 days of treatment, the removal ranged from 22.3%-30.3% and 38.6%-43.7% for imidacloprid and thiamethoxam, respectively. Transformation products imidacloprid-urea, desnitro-imidacloprid and desnitro-olefin-imidacloprid were detected in both biomixtures. The mineralization of 14C-imidacloprid revealed DT50 (mineralization half-lives) values of 3466 and 7702 days in the biomixtures B1 and B2, respectively, markedly lower than those in the soil used in their preparation (8667 and 9902 days, respectively). As demonstrated by these findings, the high persistence of these compounds in the BPS suggests that additional biological (or physicochemical) approaches should be explored in order to decrease the impact of neonicotinoid-containing wastewater of agricultural origin.

Antibiotics do not affect the degradation of fungicides and enhance the mineralization of chlorpyrifos in biomixtures.

The use of antibiotics in agriculture produces residues in wastewaters. The disposal of such wastewaters in biopurification systems (BPS) employed for the treatment of pesticides could result in the inhibition of the degrading capacity of the biomixtures used in the BPS. We assayed the effect of two commercial formulations of antibiotics used in agriculture, one containing kasugamycin (KSG) and the other oxytetracycline plus gentamicin (OTC+GTM), on the biomixture performance. Doses from 0.1mgkg-1 to 1000mgkg-1 of KSG increased the respiration of the biomixture, and low doses enhanced the mineralization rate of the insecticide 14C-chlorpyrifos. On the contrary, OTC+GTM depressed the respiration of the biomixture and the initial mineralization rate of 14C-chlorpyrifos; nonetheless, the antibiotics did not decrease overall mineralization values. The application of both formulations in the biomixture at a relevant concentration did not harm the removal of the fungicides carbendazim and metalaxyl, or their enhanced degradation; on the other hand, the biomixture was unable to dissipate tebuconazol or triadimenol, a result that was unchanged during the addition of the antibiotic formulations. These findings reveal that wastewater containing these antibiotics do not affect the performance of BPS. However, such a response may vary depending on the type of pesticide and microbial consortium in the biomixture.

Removal of carbamates and detoxification potential in a biomixture: Fungal bioaugmentation versus traditional use.

The use of fungal bioaugmentation represents a promising way to improve the performance of biomixtures for the elimination of pesticides. The ligninolyitc fungus Trametes versicolor was employed for the removal of three carbamates (aldicarb, ALD; methomyl, MTM; and methiocarb, MTC) in defined liquid medium; in this matrix ALD and MTM showed similar half-lives (14d), nonetheless MTC exhibited a faster removal, with a half-life of 6.5d. Then the fungus was employed in the bioaugmentation of an optimized biomixture to remove the aforementioned carbamates plus carbofuran (CFN). Bioaugmented and non-bioaugmented systems removed over 99% ALD and MTM after 8d of treatment, nonetheless a slight initial delay in the removal was observed in the bioaugmented biomixtures (removal after 3d: ALD 87%/97%; MTM 86%/99%, in bioaugmented/non-bioaugmented systems). The elimination of the other carbamates was slower, but independent of the presence of the fungus: >98% for MTM after 35d and >99.5% for CFN after 22d. Though the bioaugmentation did not improve the removal capacity of the biomixture, it favored a lower production of transformation products at the first stages of the treatment, and in both cases, a marked decrease in the toxicity of the matrix was swiftly achieved along the process (from 435 to 448 TU to values <1TU in 16d).

Influence of Dopaminergic System Genetic Variation and Lifestyle Factors on Excessive Alcohol Consumption.

AIMS: To examine the role of genetic and environmental factors in the pathogenesis of alcohol dependence in a Spanish cohort of women and men. METHODS: We analyzed the relationship between 56 genetic variants in 7 genes associated with the dopaminergic reward pathway and excessive alcohol consumption. The study sample (N = 1533, of which 746 were women) consisted of 653 heavy consumers and 880 very low consumers from the Spanish subcohort of the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort. Single nucleotide polymorphisms (SNPs) were genotyped using a customized array. Lifestyle variables were also examined to assess associations between genetic and environmental factors. RESULTS: No statistically significant differences were found between cases and controls for the allele frequencies in five genes: TH, SLC18A2, DRD1, DRD3 and COMT. Conversely, some alleles of the 12 SNPs from the DRD2 locus and the 5 from the MAOA locus showed significant associations with excessive alcohol consumption. Namely, rs10891556 (DRD2) proved to be the only SNP positively correlated with excessive alcohol consumption in both sexes. DRD2 rs1800497 and rs877138 were significantly associated in men, whereas DRD2 rs17601612 and rs4936271 and MAOA rs5906898 were associated with excessive alcohol consumption in women. A correspondence analysis provided an overall lifestyle profile of excessive drinkers, who were predominantly men who smoked, had large intakes of meat, small intakes of fruit and vegetables, whose jobs did not require high education levels and who engaged in little physical activity. CONCLUSIONS: It has shown the influence of dopaminergic pathway in the genetics of alcohol dependence with differences between men and women and providing a lifestyle profile of excessive drinkers.

Molecular characterization of oral squamous cell carcinoma using targeted next-generation sequencing.

OBJECTIVES: Many genetic factors play an important role in the development of oral squamous cell carcinoma. The aim of this study was to assess the mutational profile in oral squamous cell carcinoma using formalin-fixed, paraffin-embedded tumors from a Taiwanese population by performing targeted sequencing of 26 cancer-associated genes that are frequently mutated in solid tumors. METHODS: Next-generation sequencing was performed in 50 formalin-fixed, paraffin-embedded tumor specimens obtained from patients with oral squamous cell carcinoma. Genetic alterations in the 26 cancer-associated genes were detected using a deep sequencing (>1000X) approach. RESULTS: TP53, PIK3CA, MET, APC, CDH1, and FBXW7 were most frequently mutated genes. Most remarkably, TP53 mutations and PIK3CA mutations, which accounted for 68% and 18% of tumors, respectively, were more prevalent in a Taiwanese population. Other genes including MET (4%), APC (4%), CDH1 (2%), and FBXW7 (2%) were identified in our population. CONCLUSIONS: In summary, our study shows the feasibility of performing targeted sequencing using formalin-fixed, paraffin-embedded samples. Additionally, this study also reports the mutational landscape of oral squamous cell carcinoma in the Taiwanese population. We believe that this study will shed new light on fundamental aspects in understanding the molecular pathogenesis of oral squamous cell carcinoma and may aid in the development of new targeted therapies.

Loss of the transcription factor GLI1 identifies a signaling network in the tumor microenvironment mediating KRAS oncogene-induced transformation.

Although the biological role of KRAS is clearly established in carcinogenesis, the molecular mechanisms underlying this phenomenon are not completely understood. In this study, we provide evidence of a novel signaling network regulated by the transcription factor GLI1 mediating KRAS-induced carcinogenesis. Using pancreatic cancer (a disease with high prevalence of KRAS mutations) as a model, we show that loss of GLI1 blocks the progression of KRAS-induced pancreatic preneoplastic lesions in mice with pancreas-specific Cre-activated oncogenic mutant kras. Mice lacking GLI1 develop only low-grade lesions at low frequency, and in most cases, the pancreata are histologically normal. Further characterization of the phenotype showed a decrease in the activation of STAT3 in pancreatic preneoplastic lesions; STAT3 is a transcription factor required for the development of premalignant lesions and their progression into pancreatic cancer. Analysis of the mechanisms revealed a key role for GLI1 in maintaining the levels of activated STAT3 through the modulation of IL-6 signaling. GLI1 binds to the IL-6 mouse promoter and regulates the activity and expression of this cytokine. This newly identified GLI1/IL-6 axis is active in fibroblasts, a known source of IL-6 in the tumor microenvironment. Sonic hedgehog induces GLI1 binding to the IL-6 promoter and increases IL-6 expression in fibroblasts in a paracrine manner. Finally, we demonstrate that mutant KRAS initiates this cascade by inducing the expression of Sonic hedgehog in cancer cells. Collectively, these results define a novel role for GLI1 in carcinogenesis acting as a downstream effector of oncogenic KRAS in the tumor microenvironment.

Biological treatment of pesticide-containing wastewater from coffee crops: selection and optimization of a biomixture and biobed design.

The biopurification systems (BPS) or biobeds are employed for the treatment of pesticide-containing wastewater of agricultural origin. The use of these devices for pesticide removal requires the proper optimization of the composition of biomixtures (BPS active matrix) according to the target pesticides applied on a specific crop and the available materials used in their elaboration. This work aims to design a biomixture for the simultaneous treatment of several pesticides applied in coffee crops, according to local practices in Costa Rica. Three biomixtures containing either coffee husk, coconut fiber or rice husk (as the lignocellulosic substrate) were applied for the removal of 12 pesticides. The profiles of pesticide elimination and the mineralization of radiolabeled chlorpyrifos (14C-chlorpyrifos) revealed that the best performance was achieved with the coconut fiber biomixture, even though similar detoxification patterns were determined in every biomixture (according to immobilization in Daphnia magna and germination tests in Lactuca sativa). The optimization of this biomixture's composition by means of a central composite design permitted the definition of two optimal compositions (compost:soil:coconut fiber, % v/v) that maximized pesticide removal: i. 29:7.3:63.7 and ii. 11:7.3:81.7. The validation of these optimized compositions also included the use of an alternative soil from another coffee farm and resulted in overall DT50 values of 7.8-9.0 d for the pesticide mixture. Considering the removal kinetics in the optimized biomixture, a 1 m3 BPS prototype was dimensioned to be eventually used in local coffee farms. This work provides relevant information for the design and implementation of BPS at on-farm conditions for the treatment of pesticide-containing wastewater of a major crop.

Changes in the levels of thioredoxin and indoleamine-2,3-dioxygenase activity in plasma of patients with colorectal cancer treated with chemotherapy.

Increased oxidative stress and indoleamine-2,3-dioxygenase (IDO) activity have been reported in cancer, but their relationship with chemotherapy remains unknown. The aim of the present study was to examine wether the chemotherapy treatments used in colorectal cancer had an additional effect on oxidative stress and on IDO activity. Plasma samples were collected from 27 colorectal cancer patients on cytostatic treatment, 27 with cytostatic drugs plus monoclonal antibodies (cytostatic-Mabs) and 15 non-treated patients. All patients with colorectal cancer had high plasma malondialdehyde (MDA), thioredoxin (Trx) levels, and elevated IDO activity in plasma (IDOp) and in dendritic cells (IDOc). This study shows that treatment with cytostatics have an effect on oxidative stress by increasing MDA levels and by decreasing Trx levels and IDO activity. However, treatment with cytostatic-Mabs showed no effect on MDA levels but decreased Trx levels, and the IDO activity showed values similar to the healthy group. Significant correlations between plasma IDO activity and the levels of Trx (r = 0.2062, p < 0.05) and MDA (r = 0.2873, p < 0.005) were observed. Furthermore, our study suggests that IDO activity measured as kynurenine levels could be used as a marker of the response to the chemotherapy treatments, although further studies are necessary.

Pravastatin inhibits cell proliferation and increased MAT1A expression in hepatocarcinoma cells and in vivo models.

BACKGROUND: Statins may have therapeutic effects on hepatocarcinoma (HCC). This type of disorder is the most common malignant primary tumour in the liver. Our objective was to determine whether pravastatin had a therapeutic effect in vitro and in vivo models. METHOD: We design in vitro and in vivo model. In vitro we used PLC and determine cell proliferation. In vivo, we used and animal model to determined, PCNA and MAT1A expression and transaminases levels. RESULTS: We found that pravastatin decreases cell proliferation in vitro (cell proliferation in pravastatin group was 82%, in sorafenib group 51% and in combined group 40%) and in vivo (in pravastatin group 80%, in sorafenib group 76.4% and in combined group 72.72%). The MAT1A levels, was significantly higher in Pravastatin group (D 62%, P 94%, S 71%, P + S 91%). The transaminases levels, decreased significantly in Pravastatin group (GOT and GPT levels D 619.5 U/L; 271 U/L) (P 117.5 U/L; 43.5 U/L) (S 147 U/L; 59 U/L) (P + S 142 U/L; 59 U/L). CONCLUSION: The combination of pravastatin + sorafenib were more effective than Sorafenib alone.

No evidence for PML-RARa bcr1 fusion gene in colorectal cancer.

Colorectal cancer is the third most prevalent cancer and a leading cause of cancer death. Metastatic colorectal cancer patients are treated with anti-EGFR monoclonal antibodies in combination with chemotherapy; however, the efficiency is only 10-20% of such patients. An increasing amount of data has demonstrated that response to anti-EGFR therapies is confined to patients with KRAS and BRAF wild type tumors but still some of these patients are non responders to this treatment. The presence of oncogenic deregulation of different members of EGFR downstream signaling or crosstalk molecules could predict the lack of response in these patients. In this study, 40 wild type KRAS and BRAF colorectal tumors were analyzed to elucidate whether PML-RARa bcr1 fusion gene may play a role in colorectal carcinogenesis. Specifically we want to determine if this fusion could be responsible for the inability to respond to anti-EGFR monoclonal antibodies. Here, for the first time it is reported, that PML-RARa bcr1 fusion is not responsible for colorectal tumor development and also, this translocation is not predicting the lack of efficacy of anti-EGFR therapies in wild type KRAS and BRAF colorectal cancer patients. These results also suggest that PML-RARa is unlikely to be a promising target for adjuvant therapy in colorectal cancer patients.

KRAS assay selection: sensitivity and accuracy in clinical application.

KRAS mutation status is routinely tested before the administration of chemotherapeutic agents that target EGFR such as cetuximab and panitumumab. Various commercial assays to analyze KRAS mutational status are currently certified for in vitro diagnostic use. However, the differences in the operational characteristics of the different assays have not been investigated. Since different assays could be different in specificity, sensitivity and precision, studies to clarify the best option for an adequate determination of KRAS mutation status are necessary. Until a gold standard for testing KRAS mutation status is established, the treating physician must interpret the results of any given assay keeping this limitation in mind.

Effect of neoadjuvant chemotherapy in hepatic steatosis.

UNLABELLED: Chemotherapy drugs often produce side effects in the liver. In recent years, there has been speculation about the ability to produce hepatic steatosis in patients treated with 5-fluorouracil and oxaliplatin. This prospective study examines whether these drugs can produce steatosis in patients with neoadjuvant treatment who were operated on for liver tumors. PURPOSE: Our objective was to assess the effect of neoadjuvant chemotherapy (NAC) on the development of hepatic steatosis in the healthy liver. PATIENTS AND METHODS: This was a prospective study based on 32 patients divided into two groups. The presence of steatosis was assessed using a histological score (Kleiner classification) and a biochemical method (Folch method) for patients from both groups. RESULTS: A total of 14 patients (44%) had hepatic steatosis and half of these were in each group. The steatosis was moderate to severe (grades 2-3) in 4 patients (13%), 2 in each group. The mean levels of triglycerides in the liver were 33.38 and 29.94 mg/g in group I and group II, respectively, with the difference not being statistically significant. CONCLUSIONS: Almost half of the patients treated with NAC for liver neoplasia developed steatosis. Nevertheless, NAC does not seem to increase the risk of hepatic steatosis.

An integrative water quality evaluation in two surface water bodies from a tropical agricultural region in Cartago, Costa Rica.

A monitoring study was carried out in two micro-catchments in the Reventazón basin, in Northern Cartago, Costa Rica; pesticide occurrence and water quality were analyzed. Twelve pesticides were detected, five insecticides (chlorpyrifos, carbofuran, cypermethrin, imidacloprid, and oxamyl), four fungicides (carbendazim, imazalil, metalaxyl, and thiabendazole), and three herbicides (diuron, linuron, and terbutryn); eight of them presented risk quotients RQ >1, which implies a high risk for the environment. The water quality evaluation included fourteen physicochemical and microbiological parameters, out of which thermotolerant coliforms, nitrate, and total phosphorus exceeded a selected threshold value in every sample. Five metals were also included in the evaluation, Pb was the most frequent, followed by few detections of Cd, Cu, and Cr. Four water quality indexes (WQIs) were applied, two of them, the CCME WQI, based on physicochemical parameters, and the BMWP-CR WQI, based on benthic macroinvertebrate recount adapted to Costa Rican species, categorized all the sampling points as "bad" and "very bad" quality. This work of monitoring is important in the Latin American region, where there is a lack of information for regulation improvement and management decisions. These results showed poor management of the micro-catchments in this agricultural rural area.