Reparação da parede abdominal com membranas acelulares de pericárdio bovino - Parte I - Achados clínicos e radiológicos / Abdominal wall repair with acellular bovine pericardial membranes - Part I - Clinical and radiological findings

Introdução: Reduzindo os índices de recidiva de forma impactante, o emprego de biomateriais como "telas de reforço" na reparação de diferentes defeitos da parede abdominal tornou-se rotina quase obrigatória para o sucesso dessas reparações. A partir da década de 1990 houve a introdução de matrizes biológicas acelulares, iniciando-se assim uma nova era na reparação dos defeitos da parede abdominal. O objetivo é avaliar a funcionalidade do pericárdio bovino acelularizado em reparações da parede abdominal. Método: Trinta pacientes foram submetidos a reparação de defeitos da parede abdominal, com biopróteses acelulares de pericárdio bovino, perfazendo um total de 40 implantes anatomicamente individualizados. O seguimento médio foi de 31 meses, sendo os pacientes avaliados clinicamente e radiologicamente. Em três casos foram feitas biópsias das áreas implantadas permitindo análise histológica do material. Resultados: Não se observou recidiva das herniações em nenhum dos casos, tanto clinica como radiologicamente. Também não houve registro de hematomas, infecções ou qualquer fenômeno de natureza reacional local ou sistêmica. Radiologicamente, não foi possível visualizar as matrizes no local de implantação em qualquer dos períodos de pós-operatório analisados. Conclusão: As matrizes mostraram similaridade às demais membranas biológicas descritas na literatura internacional. Representando uma importante atualização e evolução conceitual, as membranas acelulares de pericárdio bovino podem ser incorporadas ao arsenal terapêutico nas reparações de parede abdominal.

Introduction: Reducing recurrence rates significantly, the use of biomaterials as "reinforcement meshes" in the repair of different abdominal wall defects has become an almost mandatory routine for the success of these repairs. From the 1990s onwards, acellular biological matrices were introduced, thus beginning a new era in the repair of abdominal wall defects. The objective is to evaluate the functionality of the acellularized bovine pericardium in abdominal wall repairs. Method: Thirty patients underwent repair of abdominal wall defects using acellular bovine pericardium bioprostheses, making a total of 40 anatomically individualized implants. The average follow-up was 31 months, with patients being evaluated clinically and radiologically. In three cases, biopsies were taken from the implanted areas, allowing histological analysis of the material. Results: No recurrence of herniations was observed in any of the cases, both clinically and radiologically. There were also no records of bruises, infections or any phenomenon of a local or systemic reaction nature. Radiologically, it was not possible to visualize the matrices at the implantation site in any of the postoperative periods analyzed. Conclusion: The matrices showed similarity to other biological membranes described in the international literature. Representing an important update and conceptual evolution, acellular bovine pericardial membranes can be incorporated into the therapeutic arsenal in abdominal wall repairs.

Reparação da parede abdominal com membranas acelulares de pericárdio bovino - Parte II - Análises histológicas e morfométricas / Repair of the abdominal wall with acellular bovine pericardial membranes - Part II - Histological and morphometric analyses

Introdução: Análise histológica é a principal ferramenta de avaliação de biopróteses acelulares, em sua maioria em caráter experimental. O objetivo é analisar histologicamente a matriz acelular de pericárdio bovino em reparações de parede abdominal implantada em humanos. Método: De uma série de 30 reparações com a membrana, 3 pacientes foram submetidas a revisão cirúrgica não relacionada aos implantes, aos 13, 22 e 23 meses de pós-operatório, obtendo-se biópsias das áreas previamente implantadas. Além da avaliação dos aspectos básicos de biocompatibilidade e neoformação tecidual, as lâminas foram digitalizadas e submetidas a análise computadorizada com o software ImageJ para quantificação da cinética de degradação das membranas, associada à análise da dimensão fractal das amostras. Os valores obtidos para porcentagens de membrana residual tiveram suas médias comparadas por análise de variância (ANOVA) e pelo teste T de Student não pareado, também utilizado para os valores da quantificação da dimensão fractal. Resultados: Foi demonstrada a biocompatibilidade do material, com neoformação tecidual, deposição de colágeno e tecido celularizado de aspecto normal, sem reações locais importantes. Fragmentos residuais da membrana foram quantificados em 40%±7% aos 13 meses, em 20%±6% aos 22 meses e em 17%±6% aos 23 meses de pós-operatório, com a análise da dimensão fractal indicando uma progressiva degradação dos implantes, com significância estatística entre 13 meses e as amostras tardias. Conclusão: Os resultados atestaram a funcionalidade do pericárdio bovino acelular sob diferentes níveis de estresse mecânico nas reparações da parede abdominal em humanos.

Introduction: Histological analysis is the main tool for evaluating acellular bioprostheses, mostly on an experimental basis. The objective is to histologically analyze the acellular matrix of bovine pericardium in abdominal wall repairs implanted in humans. Method: From a series of 30 repairs with the membrane, 3 patients underwent surgical revision unrelated to the implants at 13, 22, and 23 months postoperatively, obtaining biopsies of the previously implanted areas. In addition to evaluating the basic aspects of biocompatibility and tissue neoformation, the slides were digitalized and subjected to computerized analysis with the ImageJ software to quantify the kinetics of membrane degradation associated with the analysis of the fractal dimension of the samples. The values obtained for percentages of residual membrane had their means compared by analysis of variance (ANOVA) and the unpaired Student's T test, also used for the fractal dimension quantification values. Results: The biocompatibility of the material was demonstrated, with tissue neoformation, collagen deposition, and cellularized tissue with a normal appearance without important local reactions. Residual fragments of the membrane were quantified at 40%±7% at 13 months, at 20%±6% at 22 months, and at 17%±6% at 23 months postoperatively, with the analysis of the fractal dimension indicating a progressive degradation of implants, with statistical significance between 13 months and late samples. Conclusion: The results confirmed the functionality of the acellular bovine pericardium under different levels of mechanical stress in abdominal wall repairs in humans.

Aesthetic effect evaluation of the xenogeneic collagen matrix in keratinized mucosa augmentation around dental implants / 口腔疾病防治

Objective@#To evaluate the stability and aesthetic effect of a xenogeneic collagen matrix (mucograft) on achieving an adequate keratinized mucosa width (KMW) around implants and to provide a reference basis for the clinical application of xenogeneic collagen matrix materials.@*Methods@#The hospital ethics committee approved the study protocol, and the patients provided informed consent. Twenty patients with a KMW<2 mm at the buccal implant site who were treated in Binzhou Medical University Affiliated Yantai Stomatological Hospital from July 2020 to September 2022 were included, and a total of 36 implants were included. The mean age of the patients was (52.0±10.4) years, of which 18 were females and 2 were males. They were divided into a free gingival graft group (FGG, control group) and a xenogeneic collagen matrix group (test group) according to different graft materials. The incremental effect of the KMW on the buccal side of the implant and the mucosal shrinkage rate was measured at 1 month and 3 months after the operation. The mucosal scar index (MSI) was evaluated after the operation.@*Results@#At 3 months postoperatively, the KMW was (3.67 ± 1.06) mm in the control group and (2.96 ± 0.98) mm in the test group, and the difference was statistically significant (t = 2.076, P<0.05). The KMW shrinkage rate was (33.34 ± 16.30) % in the test group and (22.05 ± 15.47) % in the control group at 1 month postoperatively and (51.95 ± 12.60) % in the test group and (37.44 ± 16.30) % in the control group at 3 months postoperatively, with statistically significant differences between the two groups at the same time points (P<0.05). Three months after surgery, the test group showed significantly better outcomes than the control group in terms of the five scar indicators (scar width, scar convexity, scar color, scar trace, and overall appearance), and the difference was statistically significant (P<0.05).@*Conclusion@#Xenogeneic collagen matrix can increase the peri-implant KMW and achieve a more natural and coordinated soft tissue aesthetic effect but with a higher shrinkage rate.

The applications of the plasma matrix in the treatments of dental pulp and periapical diseases / 口腔疾病防治

@#The plasma matrix is a kind of autologous blood conduct. It has been widely used in maxillofacial tissue regeneration, skin cosmetology and some other fields. Recently, to preserve the dental pulp as well as the teeth, pulp regeneration therapy and apical surgery have become increasingly important as well as the applications of bioactive materials. As a kind of autologous bioactive material, the plasma matrix has some natural advantages as it is easy to obtain and malleable. The plasma matrix can be used in the following cases: ①pulp revascularization of young permanent teeth with open apical foramina that cannot stimulate apical bleeding; ② apical barrier surgery with bone defects and large area perforation repair with bone defects or root sidewall repair surgery; ③ apical surgeries of teeth with large area of apical lesions, with or without periodontal diseases. The plasma matrix is a product derived from our blood, and there are no obvious contraindications for its use. Several systematic reviews have shown that the plasma matrix can effectively promote the regenerative repair of dental pulp in patients with periapical diseases. However, the applications of plasma matrix are different because its characteristics are affected by different preparation methods. In addition, there is still a lack of long-term clinical researches on the plasma matrix, and the histological evidences are difficult to obtain, so a large number of in vitro and in vivo experimental studies are still needed. This article will describe the applications of different kinds of plasma matrix for dental pulp regeneration and bone tissue regeneration in apical surgeries to provide references for clinicians in indication selection and prognosis evaluation.

Effect of ANKRD49 on migration of human lung adenocarcinoma cells and its mechanism / 中国生物制品学杂志

@#Objective To study the effect of ankyrin repeat domain 49(ANKRD49)on the migration of human lung adenocarcinoma cell line NCI-H1299 and its mechanism.Methods NCI-H1299 cells were infected with lentivirus vector carrying ANKRD49 gene and shRNA targeting ANKRD49 to construct the cell models stably overexpressing and knocking down ANKRD49. Meanwhile,the control cell models infected with empty lentivirus vector and lentivirus vector with scramble sequences were constructed respectively. The expression levels of ANKRD49 mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blot. The effect of ANKRD49 on cell migration was measured by scratch test. The mRNA and protein levels of matrix metalloproteinase(MMP)-2/9 and tissue inhibitor of metalloproteinase(TIMP)-1/2 were detected by real-time fluorescence quantitative PCR and Western blot. The protein expression levels of p65,p-p65,IκBα and p-IκBα were detected by Western blot.Results The levels of ANKRD49 mRNA and protein in the ANKRD49 overexpression group were significantly higher than those in the control group(t = 70. 02 and 45. 68,respectively,each P < 0. 001). Compared with the control group,the migration ability of cells in the ANKRD49 overexpression group significantly increased at 24 h and 48 h(t = 5. 343 and 3. 282,P = 0. 005 9 and 0. 030 4,respectively);The mRNA transcription levels and protein expression levels of MMP-2 and MMP-9 significantly increased(t = 9. 304 and 6. 193,P =0. 000 7 and 0. 003 5,respectively),while the mRNA and protein expression of TIMP-1 and TIMP-2 decreased significantly(t = 3. 858 and 3. 517,P = 0. 018 2 and 0. 024 5,respectively),and the values of MMP-2/TIMP-1 and MMP-9/TIMP-2 significantly increased(t = 17. 7 and 9. 682,P < 0. 001 and < 0. 01,respectively);The expression of p-p65 and pIκBα significantly increased,the total protein levels of p65 and IκBα showed no obvious change,and the values of p-p65/p65 and p-IκBα/IκBα significantly increased(t = 3. 962 and 5. 370,P = 0. 016 7 and 0. 005 8,respectively). However,knocking down of ANKRD49 presented the opposite results.Conclusion ANKRD49 promotes the migration of NCI-H1299cells by enhan-cing the expression of MMP-2/9,the values of MMP-9/TIMP-1 and MMP-2/TIMP-2 via activating NF-κB/p65 signa-ling pathway.

Bone protective effect of sinomenine against monosodium iodoacetate induced knee and hip injury in rat model: an inflammatory pathway

Purpose: Osteoarthritis (OA) is a degenerative joint disease which is categorized via destruction of joint cartilage and it also affects the various joints, especially knees and hips. Sinomenine active phytoconstituents isolated from the stem of Sinomenium acutum and already proof anti-inflammatory effect against the arthritis model of rodent. In this experimental protocol, we scrutinized the anti-osteoarthritis effect of sinomenine against monosodium iodoacetate (MIA) induced OA in rats. Methods: MIA (3 mg/50 µL) was used for inducing the OA in the rats, and rats received the oral administration of sinomenine (2.5, 5 and 7.5 mg/kg body weight) up to the end of the experimental study (four weeks). The body and organs weight were estimated. Aggrecan, C-terminal cross-linked telopeptide of type II collagen (CTX-II), glycosaminoglycans (GCGs), monocyte chemoattractant protein-1 (MCP-1), Interferon gamma (IFN-γ), antioxidant, inflammatory cytokines, inflammatory mediators and matrix metalloproteinases (MMP) were analyzed. Results: Sinomenine significantly (P < 0.001) boosted the body weight and reduced the heart weight, but the weight of spleen and kidney remain unchanged. Sinomenine significantly (P < 0.001) reduced the level of nitric oxide, MCP-1 and improved the level of aggrecan, IFN-γ and GCGs. Sinomenine remarkably upregulated the level of glutathione, superoxide dismutase and suppressed the level of malonaldehyde. It effectually modulated the level of inflammatory cytokines and inflammatory mediators and significantly (P < 0.001) reduced the level of MMPs, like MMP-1, 2, 3, 9 and 13. Conclusions: Sinomenine is a beneficial active agent for the treatment of OA disease.

Effect of EGCG-based paste as intracanal dressing, in MMPs 2 and 9 expression in dog's periapical lesions

Abstract High expression of MMP-2 and MMP-9 in periapical lesions plays an important role in the degradation of the extracellular matrix. This study aimed to investigate the effect of epigallocatechin-3-gallate (EGCG)-based endodontic paste as an intracanal dressing on the expression of MMP-2 and MMP-9 in periapical lesions. Periapical lesions were experimentally induced in 35 mature beagle dog premolars randomly divided into healthy teeth, untreated periapical lesions, periapical lesions treated in a single session (control groups), and periapical lesions treated in two sessions with EGCG or calcium hydroxide-based pastes (experimental groups). After 120 days, specimens were obtained for histopathologic and immunofluorescence analyses to assess the expression of MMP-2 and MMP-9. The statistical analysis was performed using a p-value of 0.05. Endodontic treatment in two sessions using medication with EGCG and calcium hydroxide-based pastes provided similar repair of the apical and periapical tissues and neoformation of periodontal ligament fibers, cementum, and alveolar bone (p>0.05). The experimental groups treated in two sessions with both medications presented expression of MMP-2 and MMP-9 similar to that in healthy teeth (p>0.05), and significantly lower than teeth treated in a single session or untreated periapical lesions (p <0.001). Expression of MMP-2 and MMP-9 was observed in the cytoplasm of fibroblasts, osteoblasts, cementoblasts, cementocytes, and vascular endothelium. The use of EGCG-based endodontic paste reduced the expression of MMP-2 and MMP-9 and allowed repair of periapical lesions, similar to calcium hydroxide-based paste, and superior to treatment performed in a single session.

Resumo A alta expressão de MMP-2 e MMP-9 em lesões periapicais desempenha um papel importante na degradação da matriz extracelular. Este estudo teve como objetivo investigar o efeito de uma pasta à base de epigalocatequina-3-galato (EGCG) como curativo intracanal na expressão de MMP-2 e MMP-9 em lesões periapicais. Lesões periapicais foram induzidas experimentalmente em 35 pré-molares de cães da raça beagle, maduros, divididos aleatoriamente em dentes saudáveis, lesões periapicais não tratadas, lesões periapicais tratadas em uma única sessão e lesões periapicais tratadas em duas sessões com a pasta de EGCG ou pasta à base de hidróxido de cálcio. O operador monitorou os animais e realizou a eutanásia após 120 dias para análises histopatológicas e de imunofluorescência para avaliar a expressão de MMP-2 e MMP-9. A análise estatística foi realizada utilizando p=0,05. O tratamento endodôntico em duas sessões com pasta à base de EGCG e pasta à base de hidróxido de cálcio proporcionou níveis semelhantes de reparação dos tecidos apicais e periapicais e neoformação de fibras do ligamento periodontal, cemento e osso alveolar. Em ambos os grupos, a expressão de MMP-2 e MMP-9 foi mínima, sendo observada no citoplasma de fibroblastos, osteoblastos, cementoblastos, cementócitos e endotélio vascular. Em ambos os grupos tratados em duas sessões, a expressão de MMP-2 e MMP-9 foi semelhante à dos dentes hígidos e significativamente menor do que nas lesões periapicais tratadas em sessão única ou não tratadas (p < 0,001). O uso da pasta à base de EGCG reduziu a expressão de MMP-2 e MMP-9 e permitiu o reparo de lesões periapicais, semelhante à pasta à base de hidróxido de cálcio, e foi superior ao tratamento realizado em sessão única.

Longitudinal bond strength of a universal adhesive and chemical dentin characterization under different acid etching protocols

Abstract Objective This study aimed to analyze the longitudinal bond strength of a universal adhesive and chemically characterize the dentin substrate under different acid etching protocols. Methodology Dentin samples were etched with polyacrylic acid 25% (PAA) for 10 seconds (n=3) and phosphoric acid 32% (PA) for 15 seconds (n=3) and analyzed by Fourier transform infrared spectroscopy - attenuated total reflectance (FTIR-ATR) before and after treatment. For collagen degradation, samples (n=12) were divided into 3 groups: PAA, PA, and Deionized water (control), and analyzed by the quantity of solubilized type I collagen C-terminal cross-linked telopeptides and solubilized C-terminal peptide in relation to total protein concentration (ICTPtp and CTXtp) and by their ultimate tensile strength (UTS). For the adhesive interface analysis, dentin samples (n=72) were divided into 3 groups: PAA, PA, and Self-etch (SE), and subdivided into 2 groups: 24 h (baseline) and 1 year. The following tests were performed: microtensile bond strength (μTBS) (n=48), scanning electron microscopy (SEM) (n=12), and nanoleakage (n=12). Results The FTIR of PAA showed lower reduction of the peaks in the phosphate group when compared to PA. For ICTPtp, PA showed a significantly higher value. For CTXtp, PA and PAA groups failed to statically differ from each other. UTS was significantly lower for PA. For μTBS, storage time significantly affected bond strength. The results were unaffected by the etching protocol. For SEM, after 1 year, PA had little evidence of degradation in the upper third of the adhesive interface in comparison to the other groups. Nanoleakage showed no considerable silver impregnation after 1 year in the SE group. Conclusion The use of PAA prior to a universal adhesive (when compared to PA) represents a less aggressive type of etching to dentin. However, self-etching still seems to be the best option for universal adhesive systems that have functional monomers in their composition.

Development and evaluation of Tamil Matrix Sentence Test Performance in young adults

ABSTRACT Purpose The purpose of the study was to develop the Tamil Matrix Sentence Test (TMST) and evaluate the performance of a group of young adults with normal hearing on the developed test. The developed sentences were also administered at varying intensities to obtain a performance-intensity (PI) function. Methods A base matrix with 10 sentences containing 5 words each with a total of 50 words was used to develop the TMST. The sentences had a fixed semantic sentence structure of Tamil language in the order of noun, number, adjective, object and verb. The developed test consisted of 30 lists with 10 sentences in each list. The performance of 60 young adults with normal hearing aged 18 to 24 years across the 30 lists were compared for list equivalency. To obtain the PI function the sentences were administered on 20 young adults with normal hearing at intensities from 20 dB HL to 100 dB HL in 10 dB increments. The performance across the intensity levels were compared. Results The 30 lists of TMST were found to be acoustically equivalent. However, few lists showed significant difference in the scores obtained on them compared to the rest of the lists. The PI function revealed a saturation in performance beyond 40 dB HL. Conclusion From the results it was construed that TMST can be used to evaluate the speech identification abilities of Tamil speaking listeners. Multiple lists offer the advantage of retesting without the influence of practice or listeners memorizing the test material.

Enterococcus faecium M7AN10 probiótico em matriz alimentar para cães / Enterococcus faecium M7AN10 probiotic in food matrix for dogs / Enterococcus faecium M7AN10 probiótico en matriz alimentaria para perros

Probióticos são capazes de melhorar o equilíbrio da microbiota intestinal, trazendo benefícios ao hospedeiro. Atualmente no mercado há poucas opções de alimentos, com probióticos em sua composição, destinados a cães e gatos. Portanto, o objetivo deste trabalho foi desenvolver uma matriz alimentar canina (ração úmida) com o probiótico Enterococcus faecium M7AN10. Para tal, avaliou-se a inocuidade, atividade enzimática, atividade antimicrobiana, potencial probiótico e a viabilidade do microrganismo em matriz alimentar canina. O isolado foi considerado inócuo, pois apresentou ausência de atividade hemolítica e de gelatinase, além de ser suscetível a diversos antimicrobianos. E. faecium M7AN10 apresentou atividade proteolítica e capacidade de produção de exoplissacarídeo. Em relação a atividade antimicrobiana pelo método da estria radial, o isolado inibiu Acinetobacter sp. 1, Corynebacterium sp. 4, Micrococcus luteus 33, Micrococcus luteus 43, Micrococcus sp. 3, Micrococcus sp. 20, Micrococcus sp. 36. Além disso, E. faecium M7AN10 apresentou capacidade de autoagregação de 33,50% e resistiu de forma constante quando submetido ao trato gastrointestinal in vitro em conjunto com Lacticaseibacillus rhamnosus LB 1.5 e Lacticaseibacillus paracasei LB 6.4. O cultivo misto manteve-se viável em matriz alimentar canina durante o período de oito dias. Com base nesses resultados, o isolado E. faecium M7AN10 foi considerada uma bactéria candidata a probiótico que pode vir a ser usada como aditivo em alimento para cães.

Probiotics are capable of improving the balance of the intestinal microbiota, bringing benefits to the host. Currently, on the market, there are few food options with probiotics in their composition intended for dogs and cats. Therefore, this research aimed to develop a canine food matrix (wet food) with the probiotic Enterococcus faecium M7AN10. To this end, the harmlessness, enzymatic activity, antimicrobial activity, probiotic potential, and viability of the microorganism in the canine food matrix were evaluated. The isolate was considered harmless, as it showed no hemolytic and gelatinase activity and was susceptible to several antimicrobials. E. faecium M7AN10 showed proteolytic activity and exopolysaccharide production capacity. Regarding antimicrobial activity using the radial stria method, the isolate inhibited Acinetobacter sp. 1, Corynebacterium sp. 4, Micrococcus luteus 33, Micrococcus luteus 43, Micrococcus sp. 3, Micrococcus sp. 20, Micrococcus sp. 36. Furthermore, E. faecium M7AN10 showed a self-aggregation capacity of 33.50% and resisted consistently when subjected to the gastrointestinal tract in vitro together with Lacticaseibacillus rhamnosus LB 1.5 and Lacticaseibacillus paracasei LB 6.4. The mixed culture remained viable in a canine food matrix over eight days. Based on these results, the isolate E. faecium M7AN10 was considered a candidate bacterium for a probiotic that could be used as an additive in dog food.

Los probióticos son capaces de mejorar el equilibrio de la microbiota intestinal, aportando beneficios al huésped. Actualmente en el mercado existen pocas opciones de alimentos con probióticos en su composición, destinados a perros y gatos. Por lo tanto, el objetivo de este trabajo fue desarrollar una matriz alimentaria canina (comida húmeda) con el probiótico Enterococcus faecium M7AN10. Para ello se evaluó la inocuidad, actividad enzimática, actividad antimicrobiana, potencial probiótico y viabilidad del microorganismo en matriz alimentaria canina. El aislado fue considerado inofensivo, ya que no mostró actividad hemolítica ni gelatinasa, además de ser susceptible a varios antimicrobianos. E. faecium M7AN10 mostró actividad proteolítica y capacidad de producción de exoplisacáridos. En cuanto a la actividad antimicrobiana mediante el método de las estrías radiales, el aislado inhibió a Acinetobacter sp. 1, Corynebacterium sp. 4, Micrococcus luteus 33, Micrococcus luteus 43, Micrococcus sp. 3, Micrococcus sp. 20, Micrococcus sp. 36. Además, E. faecium M7AN10 mostró una capacidad de autoagregación del 33,50% y resistió consistentemente cuando se sometió al tracto gastrointestinal in vitro junto con Lacticaseibacillus rhamnosus LB 1.5 y Lacticaseibacillus paracasei LB 6.4. El cultivo mixto permaneció viable en una matriz de alimento canino durante un período de ocho días. Con base en estos resultados, el aislado E. faecium M7AN10 se consideró una bacteria candidata para un probiótico que podría usarse como aditivo en la comida para perros.

ENAM gene polymorphisms associated with dental anomalies in individuals with cleft lip and palate

Aim: This study aimed to investigate the occurrence of enamelin gene (ENAM) single nucleotide polymorphisms (SNP) and ENAM polymorphism association with dental anomalies (DA) in individuals with unilateral or bilateral cleft lip and palate (CLP). Methods: Saliva samples were collected from 147 individuals aged between 6 and 15 years-old, both genders, and divided into 4 groups: Group 1 (G1) - CLP and DA; Group 2 (G2) - CLP without DA; Group 3 (G3) - without CLP with DA; Group 4 (G4) - without CLP and DA. The genomic DNA was extracted from saliva samples and the following ENAM SNPs markers were genotyped: rs3796703, rs3796704, rs3796705, rs7671281, rs2609428, and rs35951442. Fisher exact and Pearson's Chi-square tests statistically analyzed the results (α=5%). Results: Individuals without CLP with DA (Group 3 - 19.2%) showed statistically higher prevalence of SNP rs2609428 heterozygotes (p=0.006) than individuals with CLP and DA (Group 1 - 0%). Individuals without CLP (10%) exhibited statistically higher prevalence of mutated heterozygotes/homozygous (p=0.028) than in individuals with CLP (1.3%). Conclusion: SNP rs2609428 marker of ENAM gene may be associated with dental anomalies in individuals without cleft lip and palate

Recubrimiento radicular de recesiones gingivales con matriz dérmica acelular / Root coverage of gingival recession with acellular dermal matrix

La recesión gingival es considerada como una deformidad o condición mucogingival, la Academia Americana de Periodontología, define a la recesión gingival como el desplazamiento del margen del tejido blando apical a la unión cemento-esmalte con la exposición de la superficie radicular. El tratamiento de las recesiones gingivales es un motivo de consulta común debido a razones estéticas, hipersensibilidad dentinaria, molestias durante el cepillado e incluso temor a la pérdida dentaria. Es una situación clínica común, 60% de la población humana tiene algún tipo de recesión gingival. Al realizar el examen clínico a paciente masculino de 55 años, se observó una recesión gingival tipo 1 (RT1) sin pérdida de inserción interproximal de la clasificación de Cairo. Se realizó el colgajo posicionado coronalmente (CPC) utilizando una matriz dérmica acelular (MDA) de origen humano OrACELL®. Se obtuvo resultado favorable en el recubrimiento de recesiones gingivales múltiples; considerándolos como una buena alternativa frente a los injertos gingivales autógenos. Concluyendo que, el uso de la matriz dérmica acelular para el tratamiento de la recesión gingival tipo 1 es una adecuada opción para el recubrimiento radicular. Se recomiendan más estudios a largo plazo para ver la estabilidad de los resultados obtenidos con la MDA (AU)

Gingival recession, considered a deformity or mucogingival condition, the American Academy of Periodontology, defines gingival recession as the exposure of the root surface resulting from migration of the gingival margin apical to the cementoenamel junction (CEJ). The treatment of gingival recessions is a common reason for consultation due to aesthetic reasons, dentin hypersensitivity, discomfort during brushing and even fear of tooth loss. It is a common clinical situation, 60% of the human population has some kind of gingival recession. Clinical examination of a 55-year-old male patient showed a type 1 gingival recession (RT1) without loss of interproximal insertion of the Cairo classification. Coronally advanced flap (CAF) was performed using an acellular dermal matrix (ADM) of human origin OrACELL®. Favorable results were obtained in the coating of multiple gingival recessions; considering them as a good alternative to autogenous gingival grafts. Concluding that, the use of the acellular dermal matrix for the treatment of gingival recession type 1, is a suitable option for root lining. Further long-term studies are recommended to see the elasticity of MDA outcomes (AU)

Mecanismos preventivos de la degradación de la capa híbrida / Preventive mechanisms of the degradation of the hybrid layer

Uno de los mayores retos de la odontología actual es lograr una adhesión duradera y estable entre los materiales de restauración y los tejidos dentales. Los protocolos adhesivos han ido cambiando a lo largo del tiempo para cumplir con dicho objetivo, tratando de mantener la integridad de la capa hibrida, por lo que se revisaron los factores que provocan la degradación de la capa hibrida y los mecanismos propuestos para prevenir esta degradación. Se realizó una investigación y recopilación de información bibliográfica especializadas en el tema, en buscadores científicos como PubMed, Scielo, Redalyc, Medigraphic y Scopus. Para los criterios de inclusión se consideraron años de publicación entre el año 2002 al 2022, enfocados en trabajos de investigación relacionados con la degradación de la interfaz de unión resina-dentina y mecanismos para prevenir esta degradación en la capa híbrida. El mecanismo mas estudiado a corto y largo plazo es la aplicación de clorhexidina, la cual se utiliza después del ácido fosfórico y antes del adhesivo, inhibe la actividad proteolítica de las metaloproteinasas de la matriz (MMPs) y retarda la degradación de las fibras colágenas, consiguiendo de esta manera una mayor vida de las restauraciones adhesivas.

One of the biggest challenges in dentistry is to achieve a long-lasting and stable bond between restorative materials and dental tissues. The adhesive protocols have been changing over time to meet this objective, trying to maintain the integrity of the hybrid layer, so the factors that cause the degradation of the hybrid layer and the mechanisms proposed to prevent this degradation were reviewed. An investigation and compilation of specialized bibliographic information on the subject was carried out, in scientific search engines such as PubMed, Scielo, Redalyc, Medigraphic, Scopus, among others, as well as books. For the inclusion criteria, years of publication between 2002 and 2022 were considered, focused on research works related to the degradation of the resin-dentin bonding interface and mechanisms to prevent this degradation in the hybrid layer. The placement of CHX is used after the application of phosphoric acid and before the adhesive, it inhibits the proteolytic activity of matrix metalloproteinases (MMPs) and the degradation of collagen fibers, thus achieving a longer life of resin dental restorations.

Demineralized dentin matrix technique - a comparison of different demineralizing solutions

Abstract This study aimed to evaluate the microstructure formed after the chemical treatment of teeth, for the development of autogenous grafts from the demineralized dentin matrix (DDM) technique, in order to identify the most efficient demineralizing solution. The specimens, originating from the root and coronal portion, were submitted to ultrasonic cleaning and drying in an oven for 1h at 100 ºC. Then, the density was determined by Archimedes' principle for each specimen, using distilled water as immersion liquid. The samples were separated into five groups: Control group: negative control, Distilled water;EDTA group: positive control, trisodium EDTA; NaOCl group: 2.5% sodium hypochlorite; HCl-0.6M group: 0.6M hydrochloric acid; and H2O2/H2SO4 group: hydrogen peroxide and sulfuric acid. Each specimen was immersed for 1h in the corresponding group descaling solution at 60 ºC. Subsequently, the mass loss and density of the treated specimens were determined by Archimedes' principle. Ultimately, the specimens of each group were characterized by microtomography, Scanning Electron Microscopy, and Energy Dispersive Spectrometry X-ray (SEM-EDS). The results demonstrated that the H2O2/H2SO4 solution allowed the formation of interconnected micropores, suggesting better pore structures for application in scaffolds, when compared to the other studied solutions.

Resumo Este estudo teve como objetivo avaliar a microestrutura formada após o tratamento químico em dentes, para o desenvolvimento de enxertos autógenos a partir da técnica de matriz de dentina desmineralizada (DDM), a fim de identificar a solução desmineralizante mais eficiente. Os espécimes, provenientes da raiz e porção coronal, foram submetidos à limpeza ultrassônica e secagem em estufa por 1h a 100 ºC. Em seguida, a densidade foi determinada pelo princípio de Arquimedes para cada espécime, utilizando água destilada como líquido de imersão. As amostras foram separadas em cinco grupos: Controle: controle negativo, Água destilada; EDTA: controle positivo, EDTA trissódico; NaOCl: hipoclorito de sódio 2,5%; HCl-0.6M: ácido clorídrico 0,6M; e H2O2/H2SO4: peróxido de hidrogênio e ácido sulfúrico. Cada espécime foi imerso por 1h na solução descalcificante de grupo correspondente a 60 ºC. Posteriormente, a perda de massa e a densidade dos espécimes tratados foram determinadas pelo princípio de Arquimedes. Por fim, os espécimes de cada grupo foram caracterizados por microtomografia, microscopia eletrônica de varredura e espectrometria de energia dispersiva de raios-X (SEM-EDS). Os resultados demonstraram que a solução H2O2/H2SO4 permitiu a formação de microporos interligados, sugerindo melhores estruturas de poros para aplicação em scaffolds, quando comparada às demais soluções estudadas.

Fusariosis en pacientes con cáncer: serie de 13 casos y revisión de la literatura / Fusariosis in cancer patients: 13 case series report and literature review

La fusariosis es una micosis oportunista producida por Fusarium spp. Su presentación clínica depende del estado inmunológico del huésped, especialmente, el de aquellos con enfermedades hematooncológicas, cuyas manifestaciones varían desde formas localizadas hasta infección fúngica invasora. El cultivo de piel o de sangre permite orientar el tratamiento antifúngico combinado con anfotericina B y voriconazol. Se presentan 13 casos de pacientes con cáncer en un periodo de once años que desarrollaron fusariosis diseminada; asimismo, se hizo con una revisión extensa de la literatura. En esta serie de casos, la mortalidad fue del 61,5 % (8/13), a pesar del uso del antifúngico. De los 13 pacientes, 11 tenían neoplasia hematológica y 2 neoplasia sólida. El factor de riesgo más importante fue la neutropenia profunda. El compromiso de la piel y los hemocultivos positivos facilitaron la prescripción del tratamiento combinado en la mayoría de los casos. La neutropenia febril persistente asociada a lesiones cutáneas, la onicomicosis, los nódulos o las masas pulmonares permitieron sospechar una infección fúngica invasora por Fusarium spp. El objetivo de la presentación de esta serie de casos es recordar el diagnóstico de fusariosis a la comunidad médica en contacto con pacientes oncológicos, con neutropenia febril profunda y persistentes.

The fusariosis is an opportunistic mycosis caused by Fusarium spp. Its clinical presentation depends on the immunological status of the host, especially in patients with hemato-oncological diseases, whose manifestations vary from localized to invasive fungal infections. Skin or blood culture helps to guide combined antifungal treatment with amphotericin B and voriconazole. Here, we present 13 cases in a period of eleven years of patients with cancer who developed disseminated fusariosis and their outcomes, together with a review of the related literature. In this series of cases, mortality was 61.5 % (8/13), despite the use of the antifungal. Out of the 13 cases, 11 had hematological neoplasia and 2 solid neoplasia. The most determinant risk factor was profound neutropenia. Skin involvement and positive blood cultures in most cases allowed combined treatment prescription. Persistent febrile neutropenia associated with skin lesions, onychomycosis, nodules, or lung masses lead to suspicion of Fusarium spp. fungal invasive infection. The aim of this series of cases is to remind healthcare professionals that oncological patients with deep and persistent febrile neutropenia can develop fusariosis.

Geotricosis: fungemia en paciente con leucemia linfoblástica aguda / Geotrichosis: fungemia in a patient with acute lymphoblastic leukemia

La fungemia por Geotrichum spp. es poco frecuente y altamente letal. En el Instituto Nacional de Cancerología de Bogotá solo se han reportado dos casos: uno entre el 2001 y el 2007, y el otro entre el 2012 y el 2018. Este tipo de infección es más común en pacientes con algún grado de compromiso del sistema inmunitario, por lo que puede presentarse en pacientes con neoplasias hematológicas malignas. Se presenta el caso de un hombre de 27 años con recaída de leucemia linfoblástica aguda, que ingresó con poliartralgias de cinco días de duración. También cursaba con neutropenia febril, celulitis sin abscesos y bacteriemia por Staphylococcus aureus resistente a la meticilina para lo cual recibió terapia con oxacilina y cefepime. Sin embargo, persistía la neutropenia febril por lo que se sospechó una infección fúngica invasora. Se tomó un nuevo set de hemocultivos y se inició tratamiento antifúngico. En los hemocultivos se identificaron artroconidias y mediante espectrometría de masas por láser de matriz asistida de ionización-desorción se confirmó la presencia de Geotrichum spp. Se ajustó el tratamiento antifúngico con deoxicolato de anfotericina B por 14 días y voriconazol por cuatro semanas. Luego de una estancia prolongada se le dio de alta. Aunque la incidencia de la fungemia por Geotrichum spp. es baja, en pacientes con neoplasias hematológicas malignas debe considerarse en el contexto de una neutropenia febril que es persistente a pesar del tratamiento antimicrobiano de amplio espectro. La identificación de los agentes causantes de fungemias con herramientas de proteómica, como la espectrometría de masas mencionada, permite ajustar el tratamiento dirigido y reducir las complicaciones, la estancia hospitalaria y la mortalidad.

Fungemia caused by Geotrichum spp. is rare and highly lethal. The Instituto Nacional de Cancerología in Bogotá reported just two cases: one in the period 2001-2007 and the other in 2012-2018. This type of infection is more common in any kind of immunocompromised patients, so it can occur in those with hematological malignancies. Here we present the case of a 27-year-old man, diagnosed with acute lymphoblastic leukemia in relapse and admitted with polyarthralgia for five days, febrile neutropenia, non- abscessed cellulitis, and bacteremia due to methicillin-sensitive Staphylococcus aureus. The patient received therapy with oxacillin and cefepime, but the febrile neutropenia persisted. A new set of blood cultures was taken, and antifungal treatment was started because of the suspicion of invasive fungal infection. Arthroconidia were identified in blood cultures and Geotrichum spp. was confirmed using matrix-assisted laser desorption-ionization mass spectrometry. The antifungal treatment was adjusted with amphotericin B deoxycholate for 14 days and voriconazole for four weeks, and after a prolonged stay, the patient was discharged. Although the incidence of fungemia caused by Geotrichum spp. is low, it must be considered in patients with hematological malignancies and persistent febrile neutropenia despite the broadspectrum antimicrobial treatment. The confirmation of fungemia causing agents, with proteomic tools such as the mentioned mass spectrometry, allows treatment adjustment and decreases complications, hospital stay, and mortality.

Evaluación de la dinámica de impregnación al vacío de cogollos de palma de iraca

Aunque los cogollos de la palma de iraca pueden ser empleados como fuente para la alimentación humana, la mayoría de la información disponible es para la producción de artesanías. Se evaluó la respuesta a la impregnación al vacío (IV) de cogollos frescos de palma de iraca (CFPI), con una solución isotónica de NaCl (0,6 %). Se utilizó la metodología de superficie de respuesta, con un diseño central compuesto ((=1), considerando las siguientes variables independientes: el diámetro de los cogollos (10-15 mm), el tiempo en la etapa de vacío T1 (3-5 minutos) a presión de vacío (4.1 kPa) y el tiempo en la etapa a presión atmosférica local (85,32 kPa), T2 (3-5 min). Las variables dependientes que se tomaron en cuenta fueron fracción volumétrica de impregnación en la etapa de vacío (X1), deformación volumétrica final (g), fracción volumétrica final (X) y porosidad eficaz (Ee). La dinámica de la IV del CFPI identificó que el proceso comporta una expansión volumétrica en la matriz, la cual, finalmente, contribuye a la transferencia de masa del líquido isotónico al interior de la estructura. La microestructura porosa del CFPI es compatible con el proceso de IV, permitiendo los siguientes parámetros de impregnación: g1 (0,451%), X1 (11,457%), g (2,569%), X (17,386%) y Ee (17,036%). La respuesta a la IV en los CFPI identifica a esta matriz alimentaria como adecuada, para la incorporación de componentes fisiológicamente activos.

Although the buds of the iraca palm could be employed as a source of human food, most of the information available is to produce handicrafts. The response to vacuum impregnation (VI) was evaluated in fresh iraca palm buds (FIPB) with an isotonic NaCl solution (0.6 %). The response surface methodology was obtained, with a central compound design (a= 1), considering the following independent variables: The diameter of the buds (10-15 mm), the time in the vacuum stage T1 (3-5 minutes) at vacuum pressure (4.1 kPa) and the time in the stage at local atmospheric pressure (85.32 kPa), T2 (3-5 min). The dependent variables considered were the volumetric fraction of impregnation in the vacuum stage (X1), the final volumetric deformation (g), the final volumetric fraction (X), and the effective porosity (Ee). FIPB VI dynamics identified that the process involves a volumetric expansion in the matrix, which ultimately contributes to the mass transfer of the isotonic liquid into the structure. The porous microstructure of the FIPB is compatible with the VI process, allowing the following impregnation parameters: g1 (0.451 %), X1 (11.457 %), g (2.569 %), X (17.386%), and Ee (17.036 %). The response to VI in FIPB identifies this food matrix as suitable for the incorporation of physiologically active components.

Uma clínica em extensão: a psicanálise na atenção básica / Une clinique d'extension: la psychanalyse dans les soins primaires / An extension clinic: psychoanalysis in primary care

Nas últimas décadas, observamos que a psicanálise expandiu suas fronteiras para além dos seus espaços tradicionais, como em núcleos de ensino e pesquisa universitários e serviços e movimentos psiquiátricos e de saúde mental, dividindo-se em várias correntes e em diversas associações e escolas psicanalíticas. O percurso da disciplina psicanalítica nos mostra que vários desafios foram impostos à psicanálise, dentre estes a necessidade de problematizar os campos de intervenção possíveis. O psicanalista que atua no campo institucional, a partir do seu trabalho com pacientes e equipes de saúde, deve se responsabilizar pela transmissão do saber que ali se constrói, visto que essa clínica, ao ocorrer na instituição de saúde, produz efeitos tanto para a clínica quanto para a própria instituição. Portanto, este trabalho tem por objetivo apresentar, a partir das vinhetas clínicas, as intervenções possíveis do psicanalista junto às equipes de saúde da Atenção Básica sob a perspectiva do matriciamento

Au cours des dernières décennies, nous avons observé que la psychanalyse a élargi ses frontières au-delà de ses espaces traditionnels, comme dans les centres d'enseignement et de recherche universitaire et dans les services et mouvements psychiatriques et de santé mentale, se divisant en plusieurs courants et diverses associations et écoles psychanalytiques. Le parcours de la discipline psychanalytique nous montre que plusieurs défis se sont imposés à la psychanalyse,parmi lesquels la nécessité de problématiser les champs d'intervention possibles. Le psychanalyste qui travaille dans le champ institutionnel, à partir de son travail avec les patients et les équipes soignantes, doit avoir la responsabilité de transmettre les connaissances qui s'y construisent, puisque cette clinique, lorsqu'elle se déroule dans l'établissement de santé, produit des effets tant pour la clinique que pour ainsi que pour l'institution elle-même. Ce travail vise donc présenter, à partir de vignettes cliniques, les interventions possibles du psychanalyste auprès des équipes de soins de première ligne dans une perspective d'accompagnement matriciel

In recent decades, we have observed that psychoanalysis has expanded its borders beyond its traditional spaces, such as in university teaching and research centers and psychiatric and mental health services and movements, dividing into several currents and various psychoanalytic associations and schools. The path of the psychoanalytic discipline shows us that several challenges were imposed on psychoanalysis, among them the need to problematize the possible fields of intervention. The psychoanalyst who works in the institutional field, based on his work with patients and healthcare teams, must be responsible for transmitting the knowledge that is constructed there, since this clinic, when taking place in the healthcare institution, produces effects both for the clinic as well as for the institution itself. Therefore, this work aims to present, based on clinical vignettes, the psychoanalyst's possible interventions with Primary Care health teams from the perspective of matrix support.

Demographic and population response of the threatened coral Acropora cervicornis (Scleractinia, Acroporidae) to fireworm corallivory

Introduction: The fireworm Hermodice carunculata is a widespread polychaete that can prey upon many coral species. However, few studies have examined the effect of fireworm predation on coral demographics during non-outbreak periods. Objective: To determine whether predation by H. carunculata compromised the growth, survival, and population performance of the threatened coral Acropora cervicornis. Methods: Nursery-reared coral fragments (n = 99) were fixed to the bottom of Punta Melones reef in the Island Municipality of Culebra, Puerto Rico. Predation activity and its demographic consequences on coral outplants were assessed from December 2020 to August 2022. Susceptibility to predation was compared between colonies collected directly from the reef and those originating from outside sources (e.g., coral nurseries). With the demographic data, simple size-based population matrix models were developed to 1) examine whether fireworm predation led to a significant decline in population growth rate (λ), 2) determine the demographic transition(s) that contribute the most to λ, and 3) determining the demographic transition(s) that accounted for differences in λ when comparing scenarios that considered either only predated colonies or both predated and non-predated outplants. Results: Predation increased over time, being more frequently observed in the area with the highest topographic relief and on colonies foreign to the study site. Outplants that were partially consumed grew significantly slower than non-predated colonies; however, predation did not threaten their survival. The likelihood of being attacked by the fireworm increased with branching complexity. The estimated λ for a scenario considering only predated colonies was 0.99, whereas, for a scenario where both predated and non-predated colonies were considered, λ was 0.91. Population growth, under the two scenarios, was mainly influenced by the probability of a large colony surviving and remaining at the largest size. Conclusions: Although predation can negatively impact coral growth, the relatively high survival rate of predated colonies compensates for the adverse effect. Since survival is the demographic transition that contributes most to population growth, it could be concluded that under a non-outbreak scenario, fireworm predation may not be the primary cause of A. cervicornis population decline.

Introducción: El gusano de fuego Hermodice carunculata es un poliqueto común que puede depredar muchas especies de coral. Sin embargo, pocos estudios han examinado el efecto de la depredación del gusano de fuego en la demografía de los corales durante periodos sin brotes poblacionales. Objetivo: Este estudio tuvo como objetivo determinar si la depredación por H. carunculata compromete el crecimiento, la supervivencia y el desempeño poblacional del coral amenazado Acropora cervicornis. Métodos: Fragmentos de coral criados en vivero (n = 99) se fijaron al fondo del arrecife Punta Melones en la Isla Municipio de Culebra, Puerto Rico. La actividad de depredación y sus consecuencias demográficas en los trasplantes de coral se evaluaron desde diciembre de 2020 hasta agosto de 2022. Se comparó la susceptibilidad a la depredación entre las colonias recolectadas directamente del arrecife y las que se originaron en fuentes externas (p. ej., viveros de coral). Con los datos demográficos, se desarrollaron modelos matriciales simples de población basados en el tamaño para 1) examinar si la depredación del gusano de fuego causa una disminución significativa en la tasa de crecimiento de la población (λ), 2) determinar las transiciones demográficas que más contribuyen a λ, y 3) determinar la(s) transición(es) demográfica(s) que explican las diferencias en λ al comparar escenarios que consideraron solo colonias depredadas o la combinación de colonias depredadas y no depredadas. Resultados: La depredación aumentó con el tiempo, observándose con mayor frecuencia en la zona de mayor relieve topográfico y en colonias ajenas al sitio de estudio. Los trasplantes consumidos parcialmente crecieron significativamente más lento que las colonias no depredadas; sin embargo, la depredación no amenazó su supervivencia. La probabilidad de ser atacado por el gusano de fuego aumentó con la complejidad morfológica de la colonia. El λ estimado para un escenario que consideraba solo las colonias depredadas fue de 0.99, mientras que, para un escenario en el que se consideraron tanto las colonias depredadas como las no depredadas, λ fue de 0.91. El crecimiento de la población, en ambos escenarios, estuvo influenciado principalmente por la probabilidad de que una colonia grande sobreviviera y permaneciera en el tamaño más grande. Conclusiones: Aunque la depredación puede tener un impacto negativo en el crecimiento de los corales, una tasa de supervivencia relativamente alta de las colonias depredadas compensa los efectos adversos. Dado que la supervivencia es la transición demográfica que más contribuye al crecimiento de la población, se podría concluir que, en un escenario sin brotes, la depredación por gusanos de fuego no debe ser la causa principal de la disminución de la población de A. cervicornis.

Effect of combination treatment of protocatechuic acid with 5-fluorouracil and oxaliplatin on colon cancer Caco-2 cell line

Among the most common antitumor drugs used in the treatment of colon cancer are 5-fluorouracil and oxaliplatin (5-FU and OXA). However, both these drugs have many side effects, and hence there is a need for new treatment\approach to reduce the side effects aas well as drug concentration. In this context, here, we investigated the effect of addition of protocatechuic acid (PCA) onto either monotherapies or combination therapies of 5-FU and OXA on the human colon cancer (Caco-2) cell line. In addition, we did evaluate the synergistic effect of PCA with 5-FU and OXA. Further, we determined the suppressive effects of different doses of PCA alone or in combination with 5-FU/OXA on cell proliferation after 24 and 48 hours. We identified a suppressive effect of PCA on cell viability at 48 h starting from the dose of 50 µM Matrix metalloproteinase-2 (MMP-2) and MMP-9 gene expression levels and apoptotic effects showed significant increases and decreases depending on the dose and time applied in the experimental groups. The highest synergistic activity was seen at 2:1 concentration of 5-FU+ PCA. Our findings indicate the presence of the cytotoxic and apoptotic effects of PCA in Caco-2 cells at 48 h, increasing with a dose- and time-dependent manner.