Using the Synergy between HPLC-MS and MALDI-MS Imaging to Explore the Lipidomics of Clear Cell Renal Cell Carcinoma.

Lipid imaging mass spectrometry (LIMS) has been tested in several pathological contexts, demonstrating its ability to segregate and isolate lipid signatures in complex tissues, thanks to the technique's spatial resolution. However, it cannot yet compete with the superior identification power of high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS), and therefore, very often, the latter is used to refine the assignment of the species detected by LIMS. Also, it is not clear if the differences in sensitivity and spatial resolution between the two techniques lead to a similar panel of biomarkers for a given disease. Here, we explore the capabilities of LIMS and HPLC-MS to produce a panel of lipid biomarkers to screen nephrectomy samples from 40 clear cell renal cell carcinoma patients. The same set of samples was explored by both techniques, and despite the important differences between them in terms of the number of detected and identified species (148 by LIMS and 344 by HPLC-MS in negative-ion mode) and the presence/absence of image capabilities, similar conclusions were reached: using the lipid fingerprint, it is possible to set up classifiers that correctly identify the samples as either healthy or tumor samples. The spatial resolution of LIMS enables extraction of additional information, such as the existence of necrotic areas or the existence of different tumor cell populations, but such information does not seem determinant for the correct classification of the samples, or it may be somehow compensated by the higher analytical power of HPLC-MS. Similar conclusions were reached with two very different techniques, validating their use for the discovery of lipid biomarkers.

Usefulness of an angioplasty balloon as selective bronchial blockade device after transbronchial cryobiopsy.

BACKGROUND AND OBJECTIVE: Transbronchial cryobiopsy (TBCB) is a technique in which frozen samples of lung are obtained using a probe inserted through a bronchoscope. We performed a retrospective study to assess the performance of the TBCB procedure complemented by segmental bronchial blockade using an angioplasty balloon, in terms of diagnostic yield and safety in diffuse parenchymal lung disease (DPLD). METHODS: Data from 100 patients with suspected DPLD (clinical and radiological findings), who underwent TBCB in our institution to establish a definitive diagnosis, were reviewed. In our institution, TBCB is monitored with fluoroscopy and performed under general anaesthesia by a multidisciplinary team (an anaesthesiologist, a pulmonologist and an interventional radiologist). In each patient, four samples were collected using a 2.4-mm distal diameter cryoprobe. To control bleeding, the biopsied segmental bronchus was blocked with a 6-mm diameter angioplasty balloon, inserted over a 0.035-inch angled hydrophilic guidewire. After the cryoextraction, the balloon was inflated for 3 min intervals until bleeding stopped. RESULTS: Overall, 98% of samples had diagnostic value. In 85% of cases, DPLD was confirmed, while in 7%, cancer was diagnosed. Complications were observed in 16% of the patients: 13 patients developed moderate haemorrhage, and 3 developed pneumothorax. CONCLUSION: Transbronchial cryobiopsy had a high diagnostic yield for DPLD. Performing the procedure under fluoroscopy guidance and using angioplasty balloon for selective bronchial blockade achieved a low rate of iatrogenic complications directly associated with the technique.

Quantitative analysis of the obturation of oval-shaped canals using thermoplastic techniques.

Background: One of the aim of root canal treatment is filling the root canals in a 3D way. It is not always possible to achieve due to the existence of anatomical variations. The obturation of oval canal usually provides great difficulties. Its complex anatomy can cause obstacles in instrumentation, irrigation and obturation. Aim: The main aim of this study is to quantify the area filled by gutta-percha, the area filled by cement and the area of voids present in oval canals, treated by thermoplastic filling techniques in comparison with the lateral condensation technique and observe the adaptation of the gutta-percha to the anatomy of the oval root canals according to the obturation technique. Material and Methods: 80 mandibular incisors were selected. The teeth were instrumented with Protaper Gold® and divided into 4 groups of 20. Group 1 was filled with Thermafill®; group 2 with GuttaCore®; group 3 with continuous wave vertical condensation and group 4 with the lateral condensation technique. Two horizontal sections were cut at 5mm and 7mm from the apex and they were set in putty silicone. The samples were analyzed with a Leica DMS 1000 digital microscope and processed with Leica Suite for Windows XP. The area of gutta-percha, cement and voids and the percentages, of each one, were calculated. A statistical analysis was performed using the T-Student, ANOVA tests and Kruskal-Wallis tests. Results: All thermoplastic techniques achieved a high percentage of obturation. The percentages of voids in group 4 were the highest. Statistically significant differences were found between thermoplastic techniques compared with lateral condensation. Conclusions: Thermoplastic techniques achieve better adaptation of the gutta-percha in the oval canals and low amount of cement and voids in the middle and coronal thirds respect to the lateral condensation group. Moreover, comparing termoplasthicized techniques among them, continuous wave vertical condensation got the lowest percentage of voids. Key words:Warm gutta-percha, thermoplastic obturation, oval canals, oval-shaped canals.

Characteristics and effectiveness of interventions to reduce cyberbullying: a systematic review.

Background: This paper presents the results from a systematic review on the effectiveness of interventions to reduce Cyberbullying (CB) as a function of their outcomes and main characteristics; and an analysis of the level of completeness to which the characteristics of these interventions are described. Methods: Systematic searches were conducted in PubMed, Scopus, ERIC and Psycinfo databases on February 14, 2022. In addition, relevant publications were hand searched for relevant studies. We considered interventions that provided data on CB prevalence changes in populations between primary school and college age. Results: In total, 111 studies were retained for further screening from 3,477 results. Following rigorous screening, 43 reports including 46 studies and information from 36 different interventions were included in our systematic review. Results shows that most of the interventions measuring reductions in global CB, cyberperpetration/victimization, cybervictimization and cyberperpetration were effective or partially effective. While the interventions measuring reductions in cyber-bystanding were not effective. Multicomponent interventions showed higher effectiveness than single-component interventions. After completion of the TIDieR check-list, included interventions were considered to offer an insufficient level of detail for a number of the analyzed items in relation to "how well planned," "intervention modifications" and "tailoring." Conclusion: Given the aforementioned, it is critical to increase the number of studies and the quality of interventions targeting CB and the level of detail of its description in order to obtain more robust outcomes about how to reduce its prevalence and facilitate the replication of the effective interventions. Systematic review registration: https://archive.org/details/osf-registrations-wn5u4-v1, Identifer DOI: 10.17605/OSF.IO/WN5U4.

Role of cone-beam computed tomography (CBCT) in diagnosis and treatment planning of two-rooted maxillary lateral incisor with palatogingival groove. Case report.

BACKGROUND: The embryonic root groove is an anatomical abnormality that starts in the cingulum and extends longitudinally down the long axis root towards the apex. This developmental anomaly is more frequently reported in maxillary lateral incisors. Gu YC in 2011 established three types of radicular grooves depending on its severity. According to this classification, type III presents a greater diagnostic and therapeutic complexity. The prevalence of palatogingival grooves in maxillary lateral incisors ranges from 1.9 to 14%. This case report provides valuable information about the diagnosis and treatment plan of palatogingival grooves with Cone-beam computed tomography (CBCT) scan. CASE REPORT: The patient was referred to the University Dental Clinic of European University of Valencia, with recurrent abscesses at the upper right lateral incisor region for the last two years. Palpation and percussion tests were positive for tooth 1.2. There was no clinical history of caries or previous trauma. Periapical radiography showed periapical radiolucent lesions located, not only in the apical area of tooth 1.2, but also in tooth 1.3. Both teeth had previously been endodontically treated. Periodontal probing showed normal values. CBCT scan was perfomed in order to establish a definitive diagnosis and appropriate treatment plan. DISCUSSION: The complex anatomy of the palatal root groove requires detailed knowledge of the internal root morphology for endodontic treatment success. This complementary tool allows a more accurate image of hard tissue structures, such as palatal grooves and/or accessory roots, in comparison to conventional periapical radiography. The treatment plan of this primary periodontal lesion with secondary endodontic involvement was as follows: periapical surgery combined with root amputation and sealing with MTA, and guided bone regeneration. Key words:Palatal radicular groove, palatogingival groove, maxillary lateral incisor, cone-beam computed tomography, endodontic-periodontal lesion, guided bone regeneration.

MTA apical barrier: In vitro study of the use of ultrasonic vibration.

BACKGROUND: The apexification is the first alternative treatment on a permanent tooth when, after a tooth trauma and in the presence of immature apex trauma, pulp necrosis occurs. Many studies have demonstrated the efficacy of mineral trioxide aggregate (MTA) as apical sealing material of choice in these cases, but has a degree of filtration as all other materials. The objective of this study was to analyze the seal ability of MTA on the duct walls in immature teeth unirradicular apexes, using indirect vibration. MATERIAL AND METHODS: The study was conducted on 45 teeth divided into 3 groups: Group A or control group in which no vibration for placing the MTA was used, Group B and C or groups where indirect vibration analysis was used. All samples were immersed in methylene blue to assess filtration. After performing longitudinal cuts millimetric measuring were made of the degree of filtration, divided into 3 degrees (2mm each grade filtration). RESULTS: Results obtained confirm our hypothesis, obtaining lesser degree of filtration those groups in which indirect vibration (Groups B and C) was performed. It was shown that the degree of filtration is closely linked to the degree of adaptation. CONCLUSIONS: MTA vibration offers better results in its adaptation to the canal walls, significantly reducing the degree of filtration. KEY WORDS: Apexification, MTA, filtration, indirect vibration.

Frequent loss of p53 codon 72 Pro variant in hepatitis C virus-positive carriers with hepatocellular carcinoma.

Codon 72 exon 4 polymorphism of the p53 gene has been implicated in cancer risk and it has been suggested that it may have an impact on the clinical outcome of the disease. Our objective was to evaluate the association between p53 polymorphism at codon 72 and hepatocellular carcinoma. The p53 codon 72 genotype was examined in 97 biopsy samples from 67 Basque patients histologically diagnosed with hepatocellular carcinoma. Blood samples collected from 111 Basque residents were examined as a control group. The polymorphism was examined by both single strand conformation polymorphism analysis and allele specific polymerase chain reaction. Fisher's exact test was used to evaluate the data. The results showed that there were no statistically significant differences in the frequency of codon 72 polymorphism genotype between patients with liver cancer and healthy controls. We found a frequent loss of proline allele in hepatitis C virus (HCV)-positive carriers. In conclusion, the lack of a significant relationship between this polymorphism and risk of hepatocellular carcinoma suggests that it does not predispose towards hepatocarcinogenesis in this population. We suggest that the frequent loss of the proline allele in HCV-associated carcinogenesis of the liver plays some role in hepatocarcinogenesis.

P14ARF gene alterations in human hepatocellular carcinoma.

INTRODUCTION: The molecular status of the p14(ARF) gene has not been fully elucidated in hepatocellular carcinoma (HCC). This study was performed to determine genetic and epigenetic alterations in the p14(ARF) tumor suppressor gene and their effect on HCC progression. METHODS: The status of p14 was evaluated in 117 HCC tumoral nodules and 110 corresponding non-tumor tissues by loss of heterozygosity at the 9p21-22 region, homozygous deletions, single strand conformation polymorphism-polymerase chain reaction mutational analysis and methylation-specific polymerase chain reaction. RESULTS: The most frequent inactivation mechanism was hypermethylation of the promoter region, which was found in 41.9% of tumor samples and in 19.1% of non-tumor samples. Loss of heterozygosity at the 9p21 region was detected in 27.3% and 10% of tumor and non-tumor tissues, respectively. Homozygous deletions and mutations were less common events in hepatocarcinogenesis. We found 5.9% of the tumor cases with exon 2 homozygous deletions and 3.4% of the cases with mutations. We described a silent mutation in codon 42 of exon 1beta for the first time. No association was found between inactivation of p14(ARF) and clinicopathological characteristics or prognosis. CONCLUSION: We can conclude that p14(ARF) is frequently and early altered in HCC, being the main cause of inactivation promoter hypermethylation. Our results suggest that the p14(ARF) gene plays an important role in the pathogenesis of hepatocellular carcinoma.

Overall breast density in MR mammography: diagnostic and therapeutic implications in breast cancer.

The objective of this study was to assess the efficacy of MR mammography (MRM) in evaluating breast cancer extent in women with fatty or dense breasts, and its contribution to the therapeutic approach. The authors reviewed 97 carcinomas detected in 93 women (both symptomatic and from screening) that were classified in two groups according to breast density pattern. Mammography, ultrasound (US), and MRM were performed to evaluate size, extension of the in situ component, presence of multifocal/multicentric disease, and contralateral involvement. Results obtained on mammography plus US were balanced against MRM, considering pathologic analysis as the gold standard. For fatty breasts (n=47), exact measurement was found on mammography plus US and on MRM alone in 70%, underestimation on mammography plus US 23.5% and on MRM 11% (P=0.005). For dense breasts (n=50), exact measurement was found on mammography plus US in 40% and on MRM alone 68%, underestimation on mammography plus US 52% and on MRM 10% (P=0.005). Overall, good correlation (R>0.71) was found between pathologic and clinical size with all imaging methods; nevertheless, when evaluating multifocal/multicentric disease, a poor correlation was observed between histologic assessment and mammography plus US (R=0.52), but it was excellent with regard to MRM (R=0.99). In fatty breasts, the combination of mammography and US allows for a precise assessment of tumoral extension. However, these results show that in dense breasts, MRM is superior to mammography plus US, suggesting that its systematic use in this group of patients is justifiable.

No association between GSTP1 gene aberrant promoter methylation and prognosis in surgically resected hepatocellular carcinoma patients from the Basque Country (Northern Spain).

AIMS: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, being linked etiologically to several factors. Glutathione-S-transferases (GSTs) are a family of enzymes that play an important role in detoxification. Hypermethylation of regulatory sequences at glutathione-S-transferase pi class gene (GSTP1) has been found in different human tumor types. In this study, we have studied the methylation status of the GSTP1 promoter region in patients from the Basque Country (Northern Spain) by methylation-specific PCR (MSP). METHODS AND RESULTS: GSTP1 aberrant promoter methylation was present in 24 of 117 (20.5%) tumor samples being associated with late stages of tumor progression. Patients with multiple HCCs showed different patterns of methylation, which could suggest a different clonal origin of multicentric HCC or different degrees of differentiation. No effect on disease-free survival or overall survival was observed in patients with GSTP1 methylated who underwent curative resection. CONCLUSIONS: We can conclude that GSTP1 promoter CpG island methylation appears to be a less common event during hepatocarcinogenesis in European populations than in Asian populations, being associated with late stages of tumor progression. These findings could also be useful to provide new therapeutic strategies through the use of demethylating agents.