Providing an in vitro depiction of microglial cells challenged with immunostimulatory extracellular vesicles of Naegleria fowleri.

Naegleria fowleri is the causative agent of primary amoebic meningoencephalitis, a rapid and acute infection of the central nervous system with a fatal outcome in >97% of cases. Due to the infrequent report of cases and diagnostic gaps that hinder the possibility of recovering clinic isolates, studies related to pathogenesis of the disease are scarce. However, the secretion of cytolytic molecules has been proposed as a factor involved in the progression of the infection. Several of these molecules could be included in extracellular vesicles (EVs), making them potential virulence factors and even modulators of the immune response in this infection. In this work, we evaluated the immunomodulatory effect of EVs secreted by two clinic isolates of Naegleria fowleri using in vitro models. For this purpose, characterization analyses between EVs produced by both isolates were first performed, for subsequent gene transcription analyses post incubation of these vesicles with primary cultures from mouse cell microglia and BV-2 cells. Analyses of morphological changes induced in primary culture microglia cells by the vesicles were also included, as well as the determination of the presence of nucleic acids of N. fowleri in the EV fractions. Results revealed increased expression of NOS, proinflammatory cytokines IL-6, TNF-α, and IL-23, and the regulatory cytokine IL-10 in primary cultures of microglia, as well as increased expression of NOS and IL-13 in BV-2 cells. Morphologic changes from homeostatic microglia, with small cellular body and long processes to a more amoeboid morphology were also observed after the incubation of these cells with EVs. Regarding the presence of nucleic acids, specific Naegleria fowleri DNA that could be amplified using both conventional and qPCR was confirmed in the EV fractions. Altogether, these results confirm the immunomodulatory effects of EVs of Naegleria fowleri over microglial cells and suggest a potential role of these vesicles as biomarkers of primary acute meningoencephalitis.

Efecto de la Vitamina E en Células Peritubulares de Testículos de Ratones Mus musculus Tratados con Ácido Valproico Durante el Estado Fetal y Pubertad / Effect of Vitamin E on Peritubular Cells from Testis of Mus musculus Mice Treated with Valproic Acid During the Fetal Stage and Puberty

El ácido valproico (VPA) es un fármaco antiepiléptico teratógenico que, al ser administrado durante etapas tempranas del embarazo, puede producir alteraciones en el desarrollo embriofetal, las que se manifiestan tanto a nivel del sistema nervioso como del testículo. No obstante, se ha reportado que la administración de vitamina E (VE) podría revertir dichas alteraciones. El objetivo del presente estudio fue determinar el efecto protector de la VE a nivel testicular en fetos y ratones púberes expuestos a VPA durante la fase embrionaria de su desarrollo. Se utilizó un total de 30 ratones hembra adultas gestantes (Mus musculus) cepa BALB/c, las cuales se dividieron en 6 grupos. El estudio contempló el análisis de fetos machos a los 17,5 días post-coital (dpc) y machos juveniles a las 6 semanas post-natal. A los grupos 1 y 4 se les administró 0,3 mL de solución fisiológica (grupos control para 17,5 dpc y 6 semanas postnatal, respectivamente). A los grupos 2 y 5 se les suministró la cantidad de 600 mg/kg de VPA (grupos VPA), en tanto que a los grupos 3 y 6 se les aplicó la misma dosis de VPA complementada con 200 UI de VE (grupos VPA+VE). Se describió la histología normal y patológica del compartimento peritubular del testículo. En los grupos VPA se evidenció una degeneración de la pared peritubular, y atrofia de túbulos seminíferos, así como exfoliación de las células germinales. Por el contrario, en los grupos VPA+VE tales signos no fueron observados y la morfología presentó aspecto normal solo con algunas alteraciones focales. Estos resultados corroboran el hecho que la administración de VE contrarresta en parte, los efectos deletéreos que ocasiona el VPA.

SUMMARY: Valproic acid (VPA) is a teratogenic antiepileptic drug that, when administered during the early stages of pregnancy, can produce alterations in embryo-fetal development, which manifest both at the level of the nervous system and the testicle. However, it has been reported that the administration of vitamin E (VE) could reverse these alterations. The study aimed to determine the protective effect of VE at the testicular level in fetuses and pubertal mice exposed to VPA during the embryonic phase of their development. 30 pregnant adult female mice (Mus musculus) BALB/c strain were used, which were divided into 6 groups. The study included the analysis of male fetuses at 17.5 days post-coital (dpc) and juvenile males at 6 weeks post-natal. Groups 1 and 4 were administered 0.3 mL of physiological solution. Groups 2 and 5 were given 600 mg/kg of VPA (VPA groups), while groups 3 and 6 were given the same dose of VPA supplemented with 200 IU of VE (VPA+VE). The normal and pathological histology of the peritubular compartment of the testis was described. In the VPA groups, degeneration of the peritubular wall, and atrophy of the seminiferous tubules, as well as exfoliation of the germ cells, were evident. On the contrary, in the VPA+VE groups such signs were not observed and the morphology presented a normal appearance with only some focal alterations. These results corroborate the fact that the administration of VE partially counteracts the deleterious effects caused by VPA.

Molecular orbital and topological electron density study of n → π* interactions: amides and thioamides cases.

The n → π* interactions were studied in amides and thioamides systems models, through the analysis of the electron density topology along with the Natural Bonding Orbital (NBO) approach. The effect of the dispersion terms was assessed using different DFT functionals. The NBO, independent gradient model (IGM), and the analysis of the reduced density gradient outcomes show that dispersion forces play a significant role in the strength of n → π* interactions. The IGM results indicate that δg height values for n → π* interactions do not extend beyond 0.025. All the methods used in this work predict that n → π* interaction between pairs of thioamides is stronger than those between amides. However, the electron density topology-based methods were not able to replicate the trends in the relative force of this interaction found in the experimental and NBO results.

Comparison of two handgrip dynamometers in older adults before elective surgery.

BACKGROUND: Handgrip strength is a useful measurement of muscle strength and has been proposed as a single predictor of postoperative outcomes in older adults. The aim of this study was to assess the correlation and concordance of Camry digital hand grip dynamometer (EH101) with gold standard Jamar® hydraulic handgrip dynamometer in older adults previous to elective surgery. METHODS: A cross-sectional study was conducted on patients ≥ 65 years old admitted to a Chilean private hospital for elective surgery between March 2018 and February 2019. Handgrip strength was assessed 2 times with each hand prior to surgery, using both the Jamar® dynamometer and the Camry digital dynamometer. The highest value of each dynamometer was used for analysis. RESULTS: We included a total of 220 patients (mean age 73.1 years old ± 6.3). Maximal handgrip strength averaged 26.9 kg ± 9.6 with the Camry dynamometer and 26.9 kg ± 9.7 with the Jamar® dynamometer in the right hand and 25.5 kg ± 9.5 with the Camry dynamometer and 25.7 kg ± 9.2 with the Jamar® dynamometer with the left hand. The difference between both measures did not differ significantly from 0, with Pearson correlation index of 0.95 and Lin's concordance index of 0.95 (p < 0001). The Bland-Altman graphics show that 90% of the measures were inside the confidence limits, without systematic bias. CONCLUSION: Camry digital dynamometer is an inexpensive and valid device to measure handgrip strength in older adults previous to elective surgery, compared to the gold standard Jamar® hydraulic handgrip dynamometer.

Methods for Evaluating the Effectiveness of Home Oral Hygiene Measures-A Narrative Review of Dental Biofilm Indices.

Dental plaque is a biofilm structured in an extracellular matrix of polymers of host and microbial origin; the microorganisms can coexist in harmony with the host, thus guarantying oral health. Environmental modifications can lead to dysbiosis and onset of oral diseases; in fact, plaque is the etiological agent both of periodontal disease and dental decay. The use of an effective oral hygiene index should be considered as a relevant goal for the clinicians and the researchers, and consequently, numerous plaque indices have been proposed during the years. The present literature review aims primarily to obtain a complete summary of these scores to assess plaque deposits. It is useful because the clinician/researcher will select the right scoring method for the specific situation only if he knows the available options and if he is aware of both their strengths and weaknesses. This review applies a basic classification of plaque indices that distinguishes the ones that use non-quantitative methods from the ones that use quantitative methods. Non-quantitative methods are more subjective because they are based on the ability of the clinician to point out the presence or the entity of deposits, while quantitative methods introduce objectifiable means to measure plaque deposits.

Potential of Oral Cavity Stem Cells for Bone Regeneration: A Scoping Review.

Bone loss is a common problem that ranges from small defects to large defects after trauma, surgery, or congenital malformations. The oral cavity is a rich source of mesenchymal stromal cells (MSCs). Researchers have documented their isolation and studied their osteogenic potential. Therefore, the objective of this review was to analyze and compare the potential of MSCs from the oral cavity for use in bone regeneration. METHODS: A scoping review was carried out following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) guidelines. The databases reviewed were PubMed, SCOPUS, Scientific Electronic Library Online (SciELO), and Web of Science. Studies using stem cells from the oral cavity to promote bone regeneration were included. RESULTS: A total of 726 studies were found, of which 27 were selected. The MSCs used to repair bone defects were (I) dental pulp stem cells of permanent teeth, (II) stem cells derived from inflamed dental pulp, (III) stem cells from exfoliated deciduous teeth, (IV) periodontal ligament stem cells, (V) cultured autogenous periosteal cells, (VI) buccal fat pad-derived cells, and (VII) autologous bone-derived mesenchymal stem cells. Stem cells associate with scaffolds to facilitate insertion into the bone defect and to enhance bone regeneration. The biological risk and morbidity of the MSC-grafted site were minimal. Successful bone formation after MSC grafting has been shown for small defects with stem cells from the periodontal ligament and dental pulp as well as larger defects with stem cells from the periosteum, bone, and buccal fat pad. CONCLUSIONS: Stem cells of maxillofacial origin are a promising alternative to treat small and large craniofacial bone defects; however, an additional scaffold complement is required for stem cell delivery.

Reliability assessment of the 2018 classification case definitions of peri-implant health, peri-implant mucositis, and peri-implantitis.

BACKGROUND: The purpose of this study was to evaluate the reliability and accuracy in the assignment of the case definitions of peri-implant health and diseases according to the 2018 Classification of Periodontal and Peri-implant Diseases and Conditions. METHODS: Ten undergraduate students, 10 general dentists, and 10 experts in implant dentistry participated in this study. All examiners were provided with clinical and radiographic documentation of 25 dental implants. Eleven out the 25 cases were also accompanied by baseline readings. Examiners were asked to define all cases using the 2018 classification case definitions. Reliability among examiners was evaluated using the Fleiss kappa statistic. Accuracy was estimated using percentage of complete agreement and quadratic weighted kappa for pairwise comparisons between each rater and a gold standard diagnosis. RESULTS: The Fleiss kappa was 0.50 (95% CI: 0.48 to 0.51) and the mean quadratic weighted kappa value was 0.544. Complete agreement with the gold standard diagnosis was achieved in 59.8% of the cases. Expertise in implantology affected accuracy positively (p < 0.001) while the absence of baseline readings affected it negatively (p < 0.001). CONCLUSION: Both reliability and accuracy in assigning case definitions to dental implants according to the 2018 classification were mostly moderate. Some difficulties arose in the presence of specific challenging scenarios.

Clinical, histological, immunohistochemical, and biomolecular analysis of hyaluronic acid in early wound healing of human gingival tissues: A randomized, split-mouth trial.

BACKGROUND: Hyaluronic acid (HA) exerts a fundamental role in tissue repair. In vitro and animal studies demonstrated its ability to enhance wound healing. Nevertheless, in vivo human studies evaluating mechanisms involved in oral soft tissue repair are lacking. The aim of this study was to evaluate the in vivo effect of HA on early wound healing of human gingival (G) tissues. METHODS: In the present randomized, split-mouth, double-blind, clinical trial, G biopsies were obtained in eight patients 24 h post-surgery after HA application (HA group) and compared with those obtained from the same patients without HA application (no treatment; NT group). Clinical response was evaluated through the Early Wound Healing Score (EHS). Microvascular density (MVD), collagen content and cellular proliferation were evaluated through sirius red and Masson trichrome staining, and Ki-67 immunohistochemistry, respectively. To assess collagen turnover, MMP-1, MMP-2, MMP-9, TGF-ß1 protein levels and LOX, MMP1, TIMP1, TGFB1 gene expression were analyzed by western blot and real time polymerase chain reaction. RESULTS: Twenty-four hours after surgery, the EHS was significantly higher in the HA group. MVD, collagen content, and cell proliferation were not affected. LOX mRNA, MMP-1 protein, and TIMP1 gene expression were significantly upregulated in the HA compared to the NT group. CONCLUSIONS: The additional use of 0.8% HA gel does not modify new blood vessel growth in the early phase of gingival wound healing. Concerning the secondary outcomes, HA seems to enhance extracellular matrix remodeling and collagen maturation, which could drive early wound healing of G tissues to improve clinical parameters.

Clinical effects of the adjunctive use of polynucleotide and hyaluronic acid-based gel in the subgingival re-instrumentation of residual periodontal pockets: A randomized, split-mouth clinical trial.

BACKGROUND: Polynucleotides (PN) and hyaluronic acid (HA) have been effective in stimulating the growth of primary gingival fibroblasts and promoting wound healing. The aim of this study was to investigate the clinical efficacy of a gel containing PN and HA used in association with subgingival re-instrumentation in the treatment of residual periodontal pockets. METHODS: Fifty patients were enrolled in a randomized, split-mouth, single-blind, clinical trial. For each patient, two teeth with similar residual pockets with probing depth (PD) ≥5 mm were selected to receive re-instrumentation with (test group) or without (control group) the adjunctive use of a PN and HA-based gel. Differences in changes of PD, gingival recession, clinical attachment level (CAL), modified sulcular bleeding index (mSBI), plaque index (PI) from baseline to 6, 8, 24, 36, and 48 weeks were analyzed and the frequencies of sites with PD ≤4 mm at 48 weeks were compared. RESULTS: At 48 weeks, the test group showed better results in terms of PD reduction (2.08 ± 1.24 vs. 1.94 ± 1.19, p = 0.533) and sites with PD ≤4 mm (38/50 vs. 35/50, p = 0.499), although not statistically significant. Similarly, CAL gain was comparable between groups (test: 0.50 ± 1.85 vs. CONTROL: 0.36 ± 1.80, p = 0.700). Significantly higher reduction in mSBI was recorded in the test group only in sites with baseline PD ≥6 mm (p = 0.004). CONCLUSIONS: The adjunctive use of a PN and HA-based gel could help to ensure a greater reduction of clinical parameters of inflammation in deep residual pockets.

Hyaluronic Acid as an Adjunct to Coronally Advanced Flap Procedures for Gingival Recessions: A Systematic Review and Meta-Analysis of Randomized Clinical Trials.

Previous systematic reviews have reported that coronally advanced flap (CAF) + connective tissue graft (CTG) are the gold standard in root coverage procedures (RCP). Nevertheless, adjunctive treatment with hyaluronic acid (HA) has been proposed to aim at improving clinical outcomes and reducing patient morbidity. The aim of this systematic review and meta-analysis is to compare the use of HA as an adjunctive treatment to CAF procedures in Miller class I and II (recession type 1; RT1) gingival recession (GR) defects treatment with no adjunctive/other treatments. MEDLINE, The Cochrane Central Register of Controlled Trials, Web of Science, Scopus databases and gray literature were searched up to April 2022. The primary outcome variables were mean recession coverage (MRC) and reduction of the recession depth (RecRed). Weighted mean differences and 95% confidence intervals between treatments were estimated using a random-effect mode. From 264 titles identified, 3 RCTs reporting 90 GR defects in 60 patients were included. Overall analysis of MRC and RecRed were 0.27% (p = 0.01) and 0.40 mm (p = 0.45) in favor of CAF + HA compared to CAF alone/CAF + subepithelial connective tissue graft (SCTG), respectively, with a statistically significant difference only for MRC values. Nevertheless, due to the limited number and heterogeneity of the included studies, well-performed RCTs are needed to clarify a potential advantage of HA in RCPs in the future.

Implementation of a software application in staging and grading of periodontitis cases.

OBJECTIVES: The purpose of this study was to assess the diagnostic accuracy and the inter-rater agreement among general dentists when staging and grading periodontitis cases with the aid of a software application (SA) developed by the Italian Society of Periodontology and Implantology. MATERIALS AND METHODS: Ten general dentists were asked to independently assess 25 periodontitis cases using the SA. Accuracy was estimated using quadratic weighted kappa and examiners' percentage of agreement with a reference diagnosis provided by a gold standard examiner. Inter-rater agreement was evaluated using Fleiss kappa statistics. RESULTS: The overall case definition agreed with the reference diagnosis in 53.6% of cases. The agreements for each general dentist's pairwise comparisons against the reference definition were at least substantial in 100% of cases for stage, in 70% of cases for grade and in none of the cases for extent. Fleiss kappa was 0.818, 0.608, and 0.632 for stage, extent, and grade, respectively. The study recognized possible reasons that could lead to decreased accuracy using the SA. CONCLUSIONS: Supported by the SA, general dentists have reached substantial inter-rater agreement and highly accurate assignments of stage and grade. However, complete case definitions were correctly diagnosed in slightly over half of the cases.

Study of the Mandibular Bone Microstructure and Blood Minerals Bioavailability in Rainbow Trout (Oncorhynchus mykiss, Walbaum 1792) from Freshwater.

Farmed salmonids show alterations in bone structure that result in skeletal deformities during formation, repair, and regeneration processes, with loss of mineralization at the level of the axial skeleton, mainly the head and spine, affecting their quality of life and even causing death. Despite improving factors, such as farming conditions, diets, and genetics, bone alterations appear more frequently in farmed fish than in wild fish. Thus, we used SEM-EDX, and TGA-DSC to study bone mineralization in farmed and wild rainbow trouts. As expected, we found significant differences in the nutritional parameters of farmed and wild fish (p < 0.05). Microstructural analyses indicated that farmed fish have a more robust mineral structure (p < 0.05), confirming the differences in mineralization and microstructure between both groups. However, the mechanisms regulating absorption and distribution in the organism and their effect on bone mineralization remain to be known. In our study, the combined use of techniques such as SEM-EDX and TGA-DSC allows a clearer assessment and detailed characterization beneficial to understanding the relationship between diet control and bone microstructure.

Evaluación de la mineralización en mandíbula y diente de trucha arcoíris (Oncorhynchus mykiss) mediante microscopía electrónica de barrido de presión variable acoplada a detector de espectroscopía de rayos X de energía dispersiva / Evaluation of mineralization in jaw and teeth of rainbow trout (Oncorhynchus mykiss) using variable pressure scanning electron microscopy coupled to energy-dispersive X-ray spectroscopy detector

RESUMEN: En salmonicultura se utilizan imágenes de rayos X, para evaluar la columna vertebral y las aletas, pero estas no permiten cuantificar los minerales que constituyen el hueso, para esto se utilizan otras técnicas que son destructivas. La Microscopía Electrónica de Barrido de presión variable (VP SEM) acoplada al detector espectroscopia de rayos X de energía dispersiva (EDX) acoplado, nos permite analizar la microestructura y a la vez determinar elementos químicos, porcentaje y distribución presentes en puntos específicos en una muestra. Se utilizaron 5 truchas control y 5 con deformidad mandibular, de agua dulce en etapa de smolt, se analizó la estructura y mineralización del hueso, se eligieron en promedio 6 puntos de cuantificación por región de interés y se analizaron mediante Microscopía Electrónica de Barrido (VP SEM-EDX). Los datos obtenidos muestran porcentaje en masa de Ca promedio en hueso articular de truchas fueron de 8.07 % y de 14.48 % en truchas con deformidad mandibular y control respectivamente y el porcentaje en masa promedio de P es de 4.07 % y 7.60 %, en truchas con deformidad mandibular y control respectivamente. Se identificó además otros elementos presentes en la muestra como Mg, Na, C, O, N, S, F, Zn, Al y Fe, con especial interés en el aumento de carbono en las muestras analizadas con deformidad mandibular y la presencia de aluminio en todas las muestras. La Técnica de VP SEM-EDX, permite evaluar de forma directa, sin destrucción de la muestra y con una preparación mínima de la muestra. En el hueso, la aplicación más frecuente de SEM-EDX es la medición del contenido de Ca y P y la relación que existe entre estos elementos Ca/P, en la muestra. Paralelamente la técnica nos permite la detección de otros microelementos provenientes del agua o de la alimentación y que eventualmente pueden provocar alteraciones en los peces, confirmando la hipótesis que el microanálisis elemental tiene utilidad para la salmonicultura.

SUMMARY: In salmon farming systems, X-ray images are used to evaluate the spine and fins, but these do not allow quantifying the minerals that make up the bone, for different techniques that are destructive are used. Variable Pressure Scanning Electron Microscopy (VP SEM) coupled to an Energy Dispersive X-ray spectroscopy detector (EDX), allows us to analyze the microstructure and at the same time determine chemical elements, percentages, and distribution present at specific points in a sample. Five control and five jaw deformity trout, from freshwater and in the smolt stage were used. The structure and mineralization of the bone were analyzed, an average six quantification points were chosen per region of interest (ROI) and then they were analyzed by Scanning Electron Microscopy (VP SEM-EDX). The data obtained have shown the average mass percentage of calcium in trout joint bone was 8.07 % and 14.48 % in jaw deformity and control trout, respectively; and the average mass percentage of phosphorus is 4.07 % and 7.60 %, in jaw deformity and control trout, respectively. Other elements present in the sample were also identified, such as magnesium, sodium, carbon, oxygen, nitrogen, sulphur, fluorine, zinc, aluminium, and iron, with special interest the increase of carbon in the analyzed samples with mandibular deformity and the presence of aluminum in all samples. The VP SEM-EDX Technique allows direct evaluation, without destruction of the sample and with minimal sample preparation. In bone, the most frequent application of SEM-EDX is the measurement of the content of calcium (Ca) and phosphorus (P) and the relationship that exists between these elements, calcium/phosphorus (Ca/P), in the sample. At the same time, the technique allows us to detect other microelements from water or food that can eventually cause alterations in fish, confirming the hypothesis that elemental microanalysis is useful for salmon farming.

Bioestructura de la piel de la aleta caudal del salmón del atlántico, en estado smolt y sus interacciones con el estado chalimus del Caligus rogercresseyi / Biostructure of the atlantic salmon skin of the caudal fin, in the smolt stage and its interactions with the chalimus stage of Caligus rogercresseyi

RESUMEN: Caligus rogercresseyi es un copépodo que representa uno de los principales desafíos de la industria del cultivo de salmónidos en Chile, ya que afecta profusamente a la piel. Es preciso destacar que los peces en agua dulce y estuario no son afectados, a diferencia del salmón, que desde el post-smolt resulta muy parasitado cuando es trasladado al mar. Se han realizado múltiples estudios sobre el ciclo de vida del parásito y desarrollado tratamientos químicos, físicos y mecánicos para eliminarlos. Sin embargo, a la fecha, los tratamientos no han sido eficaces, lo que produce un problema permanente para el bienestar del animal. El propósito de este estudio fue el de reconocer la bioestructura de la piel de la aleta caudal del salmón del atlántico en los sitios de la interacción con chalimus. Para esto, se utilizaron 15 post-smolt infectados con Caligus y 5 post-smolt controles, sin Caligus. Los salmones fueron aportados por Fundación Chile y la experiencia se realizó en su propio centro experimental. Una vez realizada la eutanasia, mediante sobredosis del anestésico benzocaína, se obtubieron muestras de las aletas caudales, las cuales fueron fijadas en formalina al 10%, incluidas en paraplast para realizar cortes de 5 µm de espesor y teñidas con Tricrómico de Masson y PAS. Los resultados indicaron que la piel de la aleta caudal de los post-smolt afectados presentan mayor altura de la epidermis, escasa células secretoras de mucus y solución de continuidad en la epidermis. Además, la membrana basal se descontinúa y ocurre un aumento de melanomacrófagos en la dermis.

SUMMARY: Caligus rogercresseyi is a copepod that represents one of the main challenges of the salmon farming industry in Chile, since it profusely affects the skin. It should be noted that fish in freshwater and estuaries are not affected, unlike salmon, which from post-smolt is highly parasitized when transferred to the sea. Multiple studies have been carried out on the life cycle of the parasite and chemical, physical and mechanical treatments have been developed to eliminate them. However, to date, the treatments have not been effective, which produces a permanent problem for the welfare of the animal. The purpose of this study was to recognize the biostructure of Atlantic salmon caudal fin skin at sites of interaction with chalimus. For this, 15 post-smolt infected with Caligus and 5 post-smolt controls, without Caligus, were used. The salmon were provided by Fundación Chile and the experience was carried out in its own experimental center. Once the euthanasia was carried out, by means of an overdose of the anesthetic benzocaine, samples of the caudal fins were obtained, which were fixed in 10 % formalin, included in paraplast to make 5 µm-thick sections and stained with Masson's Trichrome and PAS. The results indicated that the skin of the caudal fin of the affected post-smolt presented a greater height of the epidermis, few mucus-secreting cells and a solution of continuity in the epidermis. In addition, the basement membrane is discontinued and an increase in melanomacrophages occurs in the dermis.

Clinical Pilot Series of Non-Self-Contained Periodontal Infrabony Defects Treated with a Slowly Resorbable Bovine Pericardium Membrane in Combination with Low-Temperature-Treated Decellularized Bovine Bone Particles.

The aim of this case series was to present the clinical outcomes of non-contained intrabony periodontal defects (IPDs) treated by means of papillary preservation flaps in association with a slowly resorbable bovine pericardium membrane (BPM) and a low-temperature-treated bovine bone graft (BBG). Eight healthy, non-smoking patients (two males and six females, mean age 48 ± 8 years) with stage 3 periodontitis and at least one site with residual probing depth (PD) ≥ 6 mm associated with a non-contained IPD ≥ 3 mm were treated. Two weeks after surgery, no adverse events were observed, and an early wound healing score (EHS) of 8.1 ± 1.0 was recorded. After 1 year, the mean probing depth (PD) reduction and mean clinical attachment level gain (CAL-gain) accounted for 4.8 ± 0.7 and 3.5 ± 0.7 mm, respectively, whereas the mean gingival recession (REC) was of 1.2 ± 0.3 mm. Radiographic bone fill was observed in all cases. In conclusion, the treatment of non-contained IPDs with a slowly resorbable BPM and a low-temperature-treated BBG could be considered safe and may result in significant clinical improvements 1 year after surgery.

Effect of Chlorhexidine Digluconate in Early Wound Healing of Human Gingival Tissues. A Histological, Immunohistochemical and Biomolecular Analysis.

Chlorhexidine digluconate (CHX) is considered the gold standard for oral cavity antiseptic treatment. Nevertheless, several in vitro studies have reported detrimental effects in oral tissue repair. The aim of the present study was to evaluate the in vivo effect of post-surgical CHX mouth rinse on gingival tissue (G) 24 h after injury. G biopsies were obtained in three patients 24 h after surgery with the indication of post-surgical 0.12% CHX use and were compared with those obtained from the same patients without any antiseptic use. Changes in collagen production, cell proliferation, and apoptosis were examined by histological and Ki-67/P53 immunohistochemical analysis. Fibrotic markers (COL1A1, αSMA), proapoptotic protein (BAX) expression, and wound healing-related gene modulation (RAC1, SERPINE1, TIMP1) were analyzed by quantitative real-time PCR analysis. CHX was able to reduce cellular proliferation and increase collagen deposition, proapoptotic molecule and fibrotic marker expression, and myofibroblast differentiation, reduce expression of RAC1 and trigger expression of SERPINE1 and TIMP1, showing "scar wound healing response" pattern. This study assessed for the first time the in vivo effects of CHX on gingival tissue. The demonstration of a CHX-induced fibrotic transformation, leading to scar repair, supports the need for new post-surgical clinical protocols based on a strategic and personalized use of CHX.

Valproic acid during pregnancy decrease the number of spermatogenic cells and testicular volume in the offspring of mice: Stereological quantification.

Valproic acid (VPA) is a drug used to treat epilepsy, bipolar disorders and headaches. As a secondary effect, this antiepileptic drug can cause a decrease in androgens and gonadotropins, and dose-dependent testicular defects, such as reduction of testicular weights, sperm motility and degeneration of the seminiferous tubules. In offspring exposed to VPA, its effects have not been evaluated, so the study aimed to determine the morphological effects of the use of VPA along testicular development in mice. 30 adult female BALB/c mice were crossed and divided by age, with embryos of 12.5 days post coitum (dpc), fetuses of 17.5 dpc and male mice 6 weeks postnatal. In each case, the pregnant mouse received 600 mg/kg of VPA, making up the VPA groups, or 0.3 mL of 0.9% physiological solution for the control groups, from the beginning to the end of the pregnancy, orally.t. A morpho-quantitative analysis was carried out on the gonadal development of the male offspring. In the groups treated with VPA, at all ages studied they had lower testicular volume. At 12.5 dpc, they showed less testicular development in the form of sex cords, with fewer gonocytes and somatic cells. At 17.5 dpc, they presented greater interstitial space, fewer spermatogonial, sustentacular Sertoli, peritubular and interstitial Leydig cells. At 6 weeks postnatal, they presented fewer spermatogonia, pachytene spermatocytes, elongated spermatids, sustentacular Sertoli and interstitial Leydig cells, with statistically significant differences. In conclusion, prenatal exposure to VPA causes histopathological alterations in the offspring of mice in testicular development, from the embryonic stage to 6 weeks postnatal.

La relevancia moral del dolor de animales de experimentación y de producción / The moral relevance of pain in experimental and production animals

RESUMEN: El aspecto emocional del dolor no solo es controvertido en aves, peces y roedores, sino también en los seres humanos autoconscientes. Cuando el dolor emocional, o la insatisfacción, es vista como un mal intrínseco, evitarla o, al menos, minimizarla es un deber moral directo. El objetivo de este artículo es mostrar que no solo los modelos animales mamíferos que se utilizan en experimentación -rata (Rattus norvegicus) y ratón (Mus musculus)- sienten dolor, como reconocen universalmente las instituciones que se ocupan del bienestar animal, sino también los peces de laboratorio (Danio rerio) y los de producción (Salmo salar), y que este hecho es moralmente relevante desde el punto de vista de todas las concepciones de la moral que suelen enfrentarse en los debates morales públicos y académicos. En consecuencia, también en el caso de los peces habría que tener en cuenta indicadores de bienestar animal.

SUMMARY: The emotional aspect of pain is not only controversial in fish and rodents, but also in self-conscious humans. When pain or dissatisfaction is seen as an intrinsic evil, avoiding it or at least minimizing it is a moral duty direct. The objective of this article is to refer to pain, stress and the most widely used indices of animal welfare both in laboratory fish (Danio rerio) and production fish (Salmo salar) as well as in rat experimental animal models (Rattus norvegicus) and mouse (Mus musculus).