hsa-miR-424-5p and hsa-miR-513c-3p dysregulation mediated by IFN-γ is associated with salivary gland dysfunction in Sjögren's syndrome patients.

Salivary secretory dysfunction in SS-patients is associated with altered proteostasis, upregulation of ATF6α and components of the ERAD complex, such as SEL1L, and downregulation of XBP-1s and GRP78. Hsa-miR-424-5p is downregulated and hsa-miR-513c-3p is overexpressed in salivary glands from SS-patients. These miRNAs emerged as candidates that could regulate ATF6/SEL1L and XBP-1s/GRP78 levels, respectively. This study aimed to evaluate the effect of IFN-γ on hsa-miR-424-5p and hsa-miR-513c-3p expression and how these miRNAs regulate their targets. In labial salivary glands (LSG) biopsies from 9 SS-patients and 7 control subjects and IFN-γ-stimulated 3D-acini were analyzed. hsa-miR-424-5p and hsa-miR-513c-3p levels were measured by TaqMan assays and their localization by ISH. mRNA, protein levels, and localization of ATF6, SEL1L, HERP, XBP-1s and GRP78 were determined by qPCR, Western blot, or immunofluorescence. Functional and interaction assays were also performed. In LSGs from SS-patients and IFN-γ-stimulated 3D-acini, hsa-miR-424-5p was downregulated and ATF6α and SEL1L were upregulated. ATF6α and SEL1L were decreased after hsa-miR-424-5p overexpression, while ATF6α, SEL1L and HERP increased after hsa-miR-424-5p silencing. Interaction assays revealed that hsa-miR-424-5p targets ATF6α directly. hsa-miR-513c-3p was upregulated and XBP-1s and GRP78 were downregulated. XBP-1s and GRP78 were decreased after hsa-miR-513c-3p overexpression, while increases in XBP-1s and GRP78 were observed after hsa-miR-513c-3p silencing. Furthermore, we determined that hsa-miR-513c-3p targets XBP-1s directly. Significant correlations were found between both miRNA levels and clinical parameters. In conclusion, IFN-γ-dependent hsa-miR-424-5p and hsa-miR-513c-3p levels affect the expression of important factors involved in cellular proteostasis that control secretory function in LSG from SS-patients.

The integrated stress response is activated in the salivary glands of Sjögren's syndrome patients.

Introduction: Primary Sjögren's syndrome (SS) is an autoimmune exocrinopathy that affects the structure and function of salivary and lachrymal glands. Labial salivary gland (LSG) acinar cells from SS patients lose cellular homeostasis and experience endoplasmic reticulum and oxidative stress. The integrated cellular stress response (ISR) is an adaptive pathway essential for restoring homeostasis against various stress-inducing factors, including pro-inflammatory cytokines, and endoplasmic reticulum and oxidative stress. ISR activation leads eIF2α phosphorylation, which transiently blocks protein synthesis while allowing the ATF4 expression, which induces a gene expression program that seeks to optimize cellular recovery. PKR, HRI, GCN2, and PERK are the four sentinel stress kinases that control eIF2α phosphorylation. Dysregulation and chronic activation of ISR signaling have pathologic consequences associated with inflammation. Methods: Here, we analyzed the activation of the ISR in LSGs of SS-patients and non-SS sicca controls, determining the mRNA, protein, and phosphorylated-protein levels of key ISR components, as well as the expression of some of ATF4 targets. Moreover, we performed a qualitative characterization of the distribution of ISR components in LSGs from both groups and evaluated if their levels correlate with clinical parameters. Results: We observed that the four ISR sensors are expressed in LSGs of both groups. However, only PKR and PERK showed increased expression and/or activation in LSGs from SS-patients. eIF2α and p-eIF2α protein levels significantly increased in SS-patients; meanwhile components of the PP1c complex responsible for eIF2α dephosphorylation decreased. ATF4 mRNA levels were decreased in LSGs from SS-patients along with hypermethylation of the ATF4 promoter. Despite low mRNA levels, SS-patients showed increased levels of ATF4 protein and ATF4-target genes involved in the antioxidant response. The acinar cells of SS-patients showed increased staining intensity for PKR, p-PKR, p-PERK, p-eIF2α, ATF4, xCT, CHOP, and NRF2. Autoantibodies, focus score, and ESSDAI were correlated with p-PERK/PERK ratio and ATF4 protein levels. Discussion: In summary, the results showed an increased ISR activation in LSGs of SS-patients. The increased protein levels of ATF4 and ATF4-target genes involved in the redox homeostasis could be part of a rescue response against the various stressful conditions to which the LSGs of SS-patients are subjected and promote cell survival.

Small RNA Expression Profiling Reveals hsa-miR-181d-5p Downregulation Associated With TNF-α Overexpression in Sjögren's Syndrome Patients.

MicroRNAs (miRNAs) are small non-coding RNAs (sRNA), that alter gene expression by binding to target messenger RNAs (mRNAs) and repressing translation. Dysregulated miRNA expression has been implicated in the pathogenesis of autoimmune diseases such as Sjögren's syndrome (SS). The aim of this study was to characterize the global profile of sRNAs in labial salivary glands (LSG) from SS-patients and to validate potential miRNA candidates implicated in glandular inflammation. LSG from 21 SS-patients and 9 sicca controls were analyzed. A global next generation sequencing (NGS)-based sRNA profiling approach was employed to identify direct targets whereby differentially expressed miRNAs were predicted using bioinformatics tools. miRNA levels were validated by TaqMan and target mRNA levels were determined by quantitative real-time PCR. We also performed in vitro assays using recombinant TNF-α. NGS shows that ~30% of sRNAs were miRNAs. In comparison with samples from sicca controls, four miRNAs were found differentially expressed in LSG from SS-patients with low focus score (LFS) and 18 from SS-patients with high focus score (HFS). The miRNA with the most significant changes identified by NGS was hsa-miR-181d-5p and downregulation was confirmed by TaqMan analysis. Levels of TNF-α mRNA, a direct target of hsa-miR-181d-5p, were significantly increased and negatively correlated with hsa-miR-181d-5p presence. Moreover, positive correlations between TNF-α transcript levels, focus score, ESSDAI, and autoantibody levels were also detected. Furthermore, TNF-α stimulation decreased hsa-miR-181d-5p levels in vitro. Downregulation of hsa-miR-181d-5p in LSG from SS-patients could contribute to the glandular pro-inflammatory environment by deregulation of its direct target TNF-α. Further dissection of the pathophysiological mechanisms underlying the hsa-miR-181d-5p-mediated action in inflammatory conditions could be useful to evaluate the benefits of increasing hsa-miR-181d-5p levels for restoration of salivary gland epithelial cell architecture and function.

Identificación de macrófagos y virus Epstein-Barr en biopsias de pacientes con Síndrome de Sjögren / Identification of macrophages and Epstein-Barr virus in salivary glands of Sjögren's syndrome patients

RESUMEN El síndrome de Sjögren (SS) es una enfermedad crónica mediada inmunológicamente. La presencia de macrófagos y el virus Epstein-Barr (VEB) se ha relacionado con su desarrollo y severidad. Los macrófagos contribuyen al proceso autoinmune local y la infección viral promueve el quiebre de la auto-tolerancia. Objetivos. Identificar la presencia de Macrófagos en el infiltrado inflamatorio y VEB en células inflamatorias, correlacionándolos con las características histológicas de glándulas salivales labiales. Metodología. En biopsias de glándulas salivales labiales (8 pacientes y 7 individuos controles) se realizó inmunohistoquímica antiCD-68 para identificar macrófagos. El conteo de macrófagos y células inflamatorias se efectuó en relación a su distribución en las glándulas salivales. La presencia del virus fue evaluada mediante hibridación in situ e inmunohistoquímica para LMP1. Se utilizó el test t no pareado y de Mann-Whitney para comparar los grupos, y coeficiente de correlación de Pearson para correlacionar con parámetros histológicos. Resultados. Se observó un mayor número de macrófagos en el infiltrado inflamatorio de pacientes (p=0,001**). Los macrófagos se distribuyeron difusamente en las glándulas de controles y en los focos inflamatorios de pacientes. En ambos grupos no se detectó la presencia del virus Epstein-Barr. Conclusión. Los pacientes con síndrome de Sjögren presentaron mayor presencia de macrófagos y su incremento es a expensas del foco inflamatorio.

ABSTRACT: Sjögren's syndrome (SS) is an immunologically mediated chronic disease of complex etiopathogenesis. Macrophages and Epstein-Barr virus are among the factors related to its development and severity. Macrophages contribute to the local autoimmune process and viral infection promotes the breakdown of self-tolerance. Objectives. Identify the presence of macrophages in the inflammatory infiltrate and Epstein-Barr virus in inflammatory cells, correlating them with the histological features of labial salivary glands. Methodology. In labial salivary glands biopsies of 8 patients and 7 control individuals, anti-CD-68 immunohistochemistry was performed to identify macrophages. The macrophages and inflammatory cells were counted in relation to their distribution in the salivary glands. The presence of the virus was evaluated by in situ hybridization for viral RNA and immunohistochemistry for latent membrane protein type 1. The comparison between both groups was made using the unpaired t-test and Mann-Whitney test. The correlations with histological parameters were established with the Pearson´s correlation coefficient. Results. A greater number of macrophages was observed in the inflammatory infiltrate of SS patients (p=0,001**). Macrophages in control individuals were diffusely distributed in the gland, while, SS in patients, they were mainly located in inflammatory foci. In both groups, no inflammatory or epithelial cells infected by the Epstein-Barr virus were identified. Conclusion. Patients with Sjögren's syndrome had a greater presence of macrophages and their increase is at the expense of the inflammatory focus.

Cambios en la vía aérea después de cirugía ortognática bimaxilar / Airway change after bimaxillary orthognathic surgery

RESUMEN: La cirugía ortognática se realiza en sujetos con algún tipo de alteración esqueletal. Los movimientos maxilo mandibulares tienen impacto en la vía aérea (VA) y este aspecto se debe incorporar en la planificación quirúrgica. El objetivo de esta investigación fue determinar los cambios generados en la VA después de realizada la cirugía ortognática. Se realizó un estudio piloto incluyendo 51 sujetos con deformidad facial de clase II y clase III; se incluyeron en base al estudio del ángulo ANB y el tipo de oclusión dentaria. Se realizaron estudios con tomografía de haz cónico identificando el volumen máximo en la vía área y las áreas mínimas y máximas; además se incluyó la posición del hueso hioide y la inclinación del plano mandibular para relacionar con la morfología de la VA; para definir significancia estadística se estableció un valor de p<0,05 incluyendo las pruebas T de student y T test. Los resultados indicaron que los sujetos clase II aumentaron significativamente el volumen y áreas máximas y mínimas de la VA; los sujetos de clase III esqueletal no presentaron diferencias significativas entre la etapa pre y post quirúrgica; el hueso hioides se presentó significativamente más anterior en ambos en casos de clase II y clase III. Es posible concluir que la VA mejora sustancialmente en sujetos con clase esqueletal facial tipo II y que se mantiene sin cambios en sujetos con clase facial tipo III.

SUMMARY: Orthognathic surgery is performed in subjects with some type of skeletal alteration. Maxillomandibular movements have an impact on the airway (AW) and this aspect must be included into surgical planning. The aim of this research is to determine the changes in the AW after orthognathic surgery. A pilot study was conducted including 51 subjects with class II and class III facial deformity; they were included using the ANB angle and the type of dental occlusion. Cone beam computed tomography were performed showing the maximum volume in the airway and the minimum and maximum areas; in addition, the position of the hyoid bone and the angle of the mandibular plane were included to relate it to the morphology of the AW; to define statistical significance, a value of p<0.05 was established, including the student's t-test and the t-test. The results showed that class II subjects significantly increased the volume and maximum and minimum areas of the AW; skeletal class III subjects did not presented significant differences between the pre- and post-surgical stage; the hyoid bone was in an anterior position in both class II and class III cases. It is possible to conclude that AW improves substantially in subjects with facial class II and remains unchanged in subjects with facial class III.

Morfología ósea facial en cirugía ortognática. ¿existe tendencia hacia el avance facial? / Facial morphology in orthognathic surgery: is there a facial advance trend?

RESUMEN: El objetivo de esta investigación fue determinar los movimientos preferidos en maxila y mandíbula para obtener normalidad en morfología facial utilizando técnicas de superimposición en análisis 3D. Se realizó un estudio descriptivo para evaluar el desplazamiento óseo bimaxilar y del hueso hioides en sujetos clase facial tipo II y clase facial tipo III sometidos a cirugía ortognática. Para la superimposición se utilizó como puntos fijos Nasion - Silla - Porion y la sutura cigomática-maxilar. Estos puntos se superpusieron en CBCT pre quirúrgico y postquirúrgico y se evaluó el desplazamiento de la espina nasal anterior, Punto A, Punto B, mentón y del hueso hioides. Para la evaluación y comparación de las variables continuas antes y después de la cirugía ortognática se utilizó la prueba T de Student. Para la correlación entre las variables, se utilizó el Test de Spearman considerando un valor p<0,05 como diferencia significativa. 44 sujetos de entre 18 y 40 años de ambos sexos, fueron incluidos en esta investigación. En el 90 % de los sujetos se realizó un movimiento sagital de avance de la maxila. El movimiento sagital de avance mandibular se realizó en el 100 % de los sujetos con clase facial tipo II, mientras que el 100 % de los sujetos con clase facial tipo III se realizó se le retroceso mandibular. El hueso hioides presentó un avance en 26 de los 27 sujetos con clase facial tipo III. Es posible concluir que existe una tendencia al avance maxilar independiente de la deformidad facial.

ABSTRACT: The objective of this research was to determine the preferred movements in the maxilla and mandible to obtain normality in facial morphology using superimposition techniques in 3D analysis. A descriptive study was carried out to evaluate bimaxillary bone displacement and hyoid bone in subjects facial class II and facial class III undergoing orthognathic surgery. were used as fixed points for superimposition: Nasion (N) - Silla (S) - Porion (Po) and the zygomatic-maxillary suture (Z). These points were superimposed in pre-surgical and post- surgical CBCT and was evaluated to displacement of the anterior nasal spine, Point A, Point B, Chin and the hyoid bone. For the evaluation and comparison of continuous variables before and after orthognathic surgery, was used the Student's t test. For the correlation between the variables, the Spearman test is used, considering a p value <0.05 as a significant difference. 44 subjects between 18 and 40 years old of both sexes were included in this research. A 90% of subjects a was performed a maxillay sagittal movement. The sagittal movement of mandibular advancement was performed in 100% with facial class type II, while 100 % of the subjects with with facial class type III had a mandibular recession. The hyoid bone advanced in 26 of the 27 subjects with facial class type II. It is possible to conclude that there is a tendency for maxillary advancement, independent of facial deformity.

Type I Interferon Dependent hsa-miR-145-5p Downregulation Modulates MUC1 and TLR4 Overexpression in Salivary Glands From Sjögren's Syndrome Patients.

Sjögren's syndrome (SS) is an autoimmune disease that mainly affects salivary glands (SG) and is characterized by overactivation of the type I interferon (IFN) pathway. Type I IFNs can decrease the levels of hsa-miR-145-5p, a miRNA with anti-inflammatory roles that is downregulated in SG from SS-patients. Two relevant targets of hsa-miR-145-5p, mucin 1 (MUC1) and toll-like receptor 4 (TLR4) are overexpressed in SS-patients and contribute to SG inflammation and dysfunction. This study aimed to evaluate if hsa-miR-145-5p modulates MUC1 and TLR4 overexpression in SG from SS-patients in a type I IFN dependent manner. Labial SG (LSG) biopsies from 9 SS-patients and 6 controls were analyzed. We determined hsa-miR-145-5p levels by TaqMan assays and the mRNA levels of MUC1, TLR4, IFN-α, IFN-ß, and IFN-stimulated genes (MX1, IFIT1, IFI44, and IFI44L) by real time-PCR. We also performed in vitro assays using type I IFNs and chemically synthesized hsa-miR-145-5p mimics and inhibitors. We validated the decreased hsa-miR-145-5p levels in LSG from SS-patients, which inversely correlated with the type I IFN score, mRNA levels of IFN-ß, MUC1, TLR4, and clinical parameters of SS-patients (Ro/La autoantibodies and focus score). IFN-α or IFN-ß stimulation downregulated hsa-miR-145-5p and increased MUC1 and TLR4 mRNA levels. Hsa-miR-145-5p overexpression decreased MUC1 and TLR4 mRNA levels, while transfection with a hsa-miR-145-5p inhibitor increased mRNA levels. Our findings show that type I IFNs decrease hsa-miR-145-5p expression leading to upregulation of MUC1 and TLR4. Together, this suggests that type I interferon-dependent hsa-miR-145-5p downregulation contributes to the perpetuation of inflammation in LSG from SS-patients.

Dysfunctional mitochondria as critical players in the inflammation of autoimmune diseases: Potential role in Sjögren's syndrome.

Relevant reviews highlight the association between dysfunctional mitochondria and inflammation, but few studies address the contribution of mitochondria and mitochondria-endoplasmic reticulum (ER) contact sites (MERCs) to cellular homeostasis and inflammatory signaling. The present review outlines the important role of mitochondria in cellular homeostasis and how dysfunctional mitochondrion can release and misplace mitochondrial components (cardiolipin, mitochondrial DNA (mtDNA), and mitochondrial formylated peptides) through multiple mechanisms. These components can act as damage-associated molecular patterns (DAMPs) and induce an inflammatory response via pattern recognition receptors (PRRs). Accumulation of damaged ROS-generating mitochondria, accompanied by the release of mitochondrial DAMPs, can activate PRRs such as the NLRP3 inflammasome, TLR9, cGAS/STING, and ZBP1. This process would explain the chronic inflammation that is observed in autoimmune diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), type I diabetes (T1D), and Sjögren's syndrome. This review also provides a comprehensive overview of the importance of MERCs to mitochondrial function and morphology, cellular homeostasis, and the inflammatory response. MERCs play an important role in calcium homeostasis by mediating the transfer of calcium from the ER to the mitochondria and thereby facilitating the production of ATP. They also contribute to the synthesis and transfer of phospholipids, protein folding in the ER, mitochondrial fission, mitochondrial fusion, initiation of autophagosome formation, regulation of cell death/survival signaling, and regulation of immune responses. Therefore, alterations within MERCs could increase inflammatory signaling, modulate ER stress responses, cell homeostasis, and ultimately, the cell fate. This study shows severe ultrastructural alterations of mitochondria in salivary gland cells from Sjögren's syndrome patients for the first time, which could trigger alterations in cellular bioenergetics. This finding could explain symptoms such as fatigue and malfunction of the salivary glands in Sjögren's syndrome patients, which would contribute to the chronic inflammatory pathology of the disease. However, this is only a first step in solving this complex puzzle, and several other important factors such as changes in mitochondrial morphology, functionality, and their important contacts with other organelles require further in-depth study. Future work should focus on detecting the key milestones that are related to inflammation in patients with autoimmune diseases, such as Sjögren´s syndrome.

Mandibular Angle Contouring Using Porous Polyethylene Stock or PEEK-based Patient Specific Implants. A Critical Analysis.

ABSTRACT: Contour augmentation and mandibular angle modification surgery is becoming increasingly. The aim of this research is to compare technique and outcomes in augmentations done with standard implants or PEEK-based patient specific implants (PSI) in mandibular angle. Data from surgical planning, operative and post-operative of 6 months follow-up were revised for 21 patients who were submitted to facial surgery using a stock implant obtained from companies currently on the market or 3D implants created with CAD/CAM technology using PEEK 3D printing. Surgical time, intra-operative and post-operative complications were compared, analyzing the advantages and disadvantage of each technique. Statistical analyses using t-test and chi-squared were performed considering P value< 0.05 for statistical differences. Twelve patients were operated on with stock implants and nine patients with PSI. The surgical time was 15 minutes less for the 3D implant surgeries (P = 0.021) and intraoperatively only the stock implants needed modifications with wear and adaptation methods; post-operative infections were observed in both groups with no significant differences (P > 0.05). The 3D implants had greater levels of facial symmetry than the stock implants, although they did not present significant differences.Considering the limitations of this study, mandibular angle implants with a PEEK-based 3D CAD/CAM are efficient, stable and have a low complication rate; the CAD/CAM strategy is useful in facial surgery and can be integrated as a standard for surgical planning in facial makeover surgery.

Patient Specific Implants to Solve Structural Facial Asymmetry After Orthognathic Surgery.

ABSTRACT: Facial asymmetry is a challenge for surgeons. Some surgical strategies could be used involved soft or hard tissue of the face. The aim of this report is to show the use of patient specific implants (PSI) in a puzzle strategy based on computer aided design/computer aided manufacturer to solve a complex structural facial asymmetry after orthognathic surgery. Twenty-five-year-old male patient complain for facial asymmetry after orthognathic surgery; main deformity was related to the shape of mandibular bone in the ramus, angle, and body. After mirror image, was chose an augmentation in the right side using 2-pieces patient specific implants and the bone reduction in the vertical high of the mandibular body in the left side. Surgical technique was realized by intra oral approach installing the ramus segment at first approach and the body segment as second to obtain stability in the fitting implant-bone-implant; the left side was treated using a guide for osteotomy; after 1-year follow-up no infection or complication was observed and facial symmetry was obtained. It is possible to conclude that the puzzle technique using polyetheretherketone can be applied to obtain predictable results in a simple strategy to solve a complex problem.

Tofacitinib counteracts IL-6 overexpression induced by deficient autophagy: implications in Sjögren's syndrome.

OBJECTIVE: Altered homeostasis of salivary gland (SG) epithelial cells in Sjögren's syndrome (SS) could be the initiating factor that leads to inflammation, secretory dysfunction and autoimmunity. Autophagy is an important homeostatic mechanism, whose deficiency is associated with inflammation and accumulation of Janus kinase (JAK)-signal transducer and activator of transcription (STAT) components. We aimed to evaluate whether autophagy is altered in labial SG (LSG) epithelial cells from primary SS (pSS) patients and whether this contributes to inflammation through the JAK-STAT pathway. Furthermore, we investigated the anti-inflammatory effect of the JAK inhibitor tofacitinib in autophagy-deficient (ATG5 knockdown) three-dimensional (3D)-acini. METHODS: We analysed LSG biopsies from 12 pSS patients with low focus score and 10 controls. ATG5-deficient 3D-acini were generated and incubated with IL-6 in the presence or absence of tofacitinib. Autophagy markers, pro-inflammatory cytokine expression, and JAK-STAT pathway activation were evaluated by PCR or western blot, along with correlation analyses between the evaluated markers and clinical parameters. RESULTS: LSG from pSS patients showed increased p62 and decreased ATG5 expression, correlating negatively with increased activation of JAK-STAT pathway components (pSTAT1 and pSTAT3). Increased expression of STAT1 and IL-6 correlated with EULAR Sjögren's syndrome disease activity index and the presence of anti-Ro antibodies. ATG5-deficient 3D-acini reproduced the findings observed in LSG from pSS patients, showing increased expression of pro-inflammatory markers such as IL-6, which was reversed by tofacitinib. CONCLUSION: Decreased expression of ATG5 in LSG epithelial cells from pSS patients possibly contributes to increased inflammation associated with JAK-STAT pathway activation, as evidenced in ATG5-deficient 3D-acini. Interestingly, these results suggest that tofacitinib could be used as an anti-inflammatory agent in pSS patients.

Three-Dimensional Planning and Surgical Technique for Modified Le Fort I and Le Fort III Osteotomy in Non-Syndromic Patients.

Total midface deficiency, to include the orbits, nose, zygomas, and maxilla, can occur in both syndromic and non-syndromic individuals. The treatment with combination of Le Fort III and I osteotomy could be used and it is few reported in the literature. The aim of the study is to present how technology can make the procedure for the correction of hypoplasia of the middle third more predictable and safer. The 2 clinical cases were managed under submental intubation and using VSP that generated 3D printing of oclusal splints and cutting guides. Modified oblique Le Fort III osteotomy (MOLFIIIO) and Le Fort I osteotomy were used due to patients present large sagittal discrepancy between maxilla and mandible (18 and 17 mm). The patients presented good outcomes without complications. In this study, the authors demonstrate that non-syndromic patient could be managed safety with MOLFIII and Le Fort I osteotomies for the correction of midfacial deformities using virtual surgical planning (VSP) associated with 3D printing technique and piezoelectric surgery.

Validity of the Genourob arthrometer in the evaluation of total thickness tears of anterior cruciate ligament.

OBJECTIVE: Our purpose was to evaluate the validity and reliability of the laxity with the GNRB arthrometer in subjects with anterior cruciate ligament injury. MATERIAL AND METHOD: A diagnostic study was performed by three operators using the Genourob arthrometer, measuring the displacement of the anterior cruciate ligament. The concordance was assessed by the intraclass correlation coefficient mixed effects model, Lin correlation coefficient and graphic method from Bland-Altman. Using the anterior cruciate ligament tear as a dependent variable and the Genourob measurement as an independent variable, a logistic regression was determined. RESULTS: Obtaining the complete information of 157 knees. The measurements with the Genourob arthrometer distributed symmetrically, with mean ± standard deviation of knees with anterior cruciate ligament injury: 5.64 ± 1.72 and knees without anterior cruciate ligament injury: 3.29 ± 1.72. The ICCs as well as the LCCs were equal to or greater than 0.99. The BA showed discrepancy for a pair of observations no greater than 7.64%. The odds ratio of the knee displacement measurement for the presence of anterior cruciate ligament injury was 4.04 (95% CI: 2.59-6.32; p-value < .01) with a ROC area of 0.863 (95% CI: 0.789-0.9456). The cut-off point of the anteroposterior knee displacement located at 6.8 mm determined a sensitivity of 74.4% and specificity of 93.8%, with a Youden Index = 0.67. CONCLUSION: The Genourob arthrometer is reliable and valid to establish where laxity values correlate with total thickness tears of the anterior cruciate ligament.

Consideraciones del Leiomioma y su Variante Intraósea en Mandíbula, Algoritmo de Trabajo Según Análisis de Literatura / Considerations of Leiomyoma and its Intraosseous Variant in the Jaw, Working Algorithm According to Literature Analysis

RESUMEN: El Leiomioma es una neoplasia benigna originada en el tejido muscular liso por lo que puede manifestarse en cualquier región del cuerpo humano que contenga músculo liso, siendo el sitio más común el útero, la piel y en el tracto gastrointestinal. La escasa cantidad de tejido muscular liso en boca hace que su manifestación oral sea infrecuente, representando solo 0.06 % del total de este tipo de tumores. Los sitios más afectados suelen ser los labios, las mejillas, el paladar, la lengua y encías. Histológicamente se distinguen tres tipos: el Leiomioma Sólido, Angioleiomioma (Vascular) y Leiomioma Epitelioide (Leiomioblastoma). El origen de esta neoplasia en boca suele ser la túnica media de los vasos sanguíneos. El Leiomioma intraóseo suele ser aún menos frecuente, y con un diagnóstico diferencial complejo, con histopatología que en varias ocasiones no suele ser fácil de clasificar. Si bien es definida como un tumor benigno, su manifestación intraósea puede llegar a ser localmente agresiva y con un diagnóstico controversial, debiendo abordarse muchas veces como una neoplasia maligna. El objetivo de este artículo es presentar una revisión de la literatura de esta variante intraósea de Leiomioma situada en mandíbula, sus consideraciones clínicas y un algoritmo de tratamiento.

ABSTRACT: Leiomyoma is a benign neoplasm, the origin is the smooth muscle tissue that can be found in any area of the human body, which contains smooth muscle tissue. The most common regions it can be located, are the uterus, the skin and the gastrointestinal tract. The low quantity of muscle tissue in the mouth leads to infrequent oral manifestation, representing only 0.06 % of these tumors. The most affected regions are the lips, cheeks, palate, tongue and gums. The Histologic classification is: Solid Leiomyoma, Angioleiomyoma (vascular) and Epithelioid Leiomyoma (Leiomyoblastoma). The origin of this tumor in the mouth is the tunica media of the blood vessels. Nevertheless, and in spite of being defined as a benign tumor, it can be extremely aggressive, be subject to controversial diagnosis, and must often be treated as a malign neoplasm. The Intraosseous Leiomyoma is infrequent and presents a complicated differential diagnosis, with a histopathology that many times cannot be easily classified. The aim of this article is to present a review of intraosseous variant Leiomyoma in the mandible, the clinicians´ considerations and a treatment algorithm.

Aberrant MUC1 accumulation in salivary glands of Sjögren's syndrome patients is reversed by TUDCA in vitro.

OBJECTIVES: Xerostomia in SS patients has been associated with low quality and quantity of salivary mucins, which are fundamental for the hydration and protection of the oral mucosa. The aim of this study was to evaluate if cytokines induce aberrant mucin expression and whether tauroursodeoxycholic acid (TUDCA) is able to counteract such an anomaly. METHODS: Labial salivary glands from 16 SS patients and 15 control subjects, as well as 3D acini or human submandibular gland cells stimulated with TNF-α or IFN-γ and co-incubated with TUDCA, were analysed. mRNA and protein levels of Mucin 1 (MUC1) and MUC7 were determined by RT-qPCR and western blot, respectively. Co-immunoprecipitation and immunofluorescence assays for mucins and GRP78 [an endoplasmic reticulum (ER)-resident protein] were also performed. mRNA levels of RelA/p65 (nuclear factor-κB subunit), TNF-α, IL-1ß, IL-6, SEL1L and EDEM1 were determined by RT-qPCR, and RelA/p65 localization was evaluated by immunofluorescence. RESULTS: MUC1 is overexpressed and accumulated in the ER of labial salivary gland from SS patients, while MUC7 accumulates throughout the cytoplasm of acinar cells; however, MUC1, but not MUC7, co-precipitated with GRP78. TUDCA diminished the overexpression and aberrant accumulation of MUC1 induced by TNF-α and IFN-γ, as well as the nuclear translocation of RelA/p65, together with the expression of inflammatory and ER stress markers in 3D acini. CONCLUSION: Chronic inflammation alters the secretory process of MUC1, inducing ER stress and affecting the quality of saliva in SS patients. TUDCA showed anti-inflammatory properties decreasing aberrant MUC1 accumulation. Further studies are necessary to evaluate the potential therapeutic effect of TUDCA in restoring glandular homeostasis in SS patients.

Identification of bacterial and fungal species in human cadavers used in anatomy teaching / Identificación de especies bacterianas y fúngicas en cadáveres humanos utilizados en la enseñanza de la anatomía

Within the framework of undergraduate and postgraduate medical education, cadavers have been used to teach anatomy by dissection or by using prosected specimens. To accomplish this, an appropriated preservation process must guarantee that the cadaver is kept safe for harm, destruction, and decomposition. Embalming fluid contains fixatives, disinfectants, surfactants, buffers, salt, and water, making the cadaver safe for teaching anatomy. However, it remains unclear if there is any risk of dissemination of microorganisms during anatomy teaching, research, and dissection procedures on fixed cadavers. The purpose of this study is to identify bacterial and fungal species in fixed cadaveric material used in anatomy teaching. Samples of cadavers and anatomical sections were cultured and biochemical tests and molecular identification by polymerase chain reaction (PCR) were performed to identify the microorganisms. The results indicate that fixed cadaveric material has viable bacteria on its surfaces and almost all these correspond to gram-negative bacilli of the Enterobacteriaceae family. In conclusion, fixed cadavers could be a reservoir of bacteria. This study underscores the importance of generating safe manipulation protocols to avoid eventual contamination and disease.

Dentro del curriculum de los programas de postgrado y pregrado de las carreras de la salud, los cadáveres han sido utilizados para la enseñanza de la anatomía mediante la disección o utilizando preparados anatómicos. Para poder llevar a cabo esto, el cadáver debe pasar por un adecuado proceso de preservación; en el que se utilizan fluidos que contienen fijadores, desinfectantes, surfactantes, buffers, sal y agua, los cuales lo protegen del deterioro y la descomposición. Las soluciones fijadoras y conservadoras contienen desinfectantes, surfactantes, fijadores, buffers, sal y agua, que hacen que el cadáver sea seguro para la enseñanza de la anatomía. Sin embargo, no está claro si existe algún riesgo de diseminación de microorganismos durante la enseñanza, investigación y/o disección en estos cadáveres. El propósito del estudio es identificar especies bacterianas y/o fúngicas en material cadavérico previamente fijado, usado en la enseñanza de la anatomía. Se realizaron cultivos y técnicas de identificación molecular mediante reacción en cadena de polimerasa de muestras tomadas desde material cadavérico para identificar los microorganismos encontrados. Los resultados indican que el material cadavérico previamente fijado posee bacterias en sus superficies, la mayoría corresponde a bacilos gram negativos de la familia de las Enterobacteriaceae. En conclusión, los cadáveres previamente fijados pueden ser reservorio de bacterias. Este estudio destaca la importancia de generar protocolos de manipulación con el fin de evitar una posible contaminación y enfermedad.

Condromatosis Sinovial Primaria de la Articulación Temporomandibular con Extensión a Hueso Temporal. Reporte de un Caso y Revisión de la Literatura / Primary Synovial Chondromatosis of the Temporomandibular Joint with Temporal Bone Extension. Case Report and Literature Review

RESUMEN: La condromatosis sinovial (CS), es una lesión benigna poco frecuente y de clínica bastante inespecífica. Suele afectar articulaciones de huesos largos como la rodilla, el codo y la cadera, presentándose generalmente de manera unilateral. Se cree que solo un 3 % de los casos de CS afecta la articulación temporomandibular. Esta condición se caracteriza por ser un trastorno metaplásico del tejido conectivo sinovial que suele manifestarse con la formación de pequeños y múltiples nódulos de cartílago que posteriormente pueden desprenderse, calcificarse y formar cuerpos libres dentro del espacio articular. Presentamos el caso de una mujer de 55 años con condromatosis sinovial de la articulación temporomandibular, tratada desde hace 3 años bajo el diagnóstico de desórdenes temporomandibulares. A pesar de ser considerada una lesión de tipo benigna, esta puede llegar a ser localmente agresiva, extendiéndose como en nuestro reporte hacia la fosa craneal media, adelgazando parte del hueso temporal.

ABSTRACT: Synovial chondromatosis (CS) is a benign lesion that is rare and clinically quite nonspecific. It usually affects the joints of long bones such as the knee, elbow and hip, usually occurring unilaterally. It is believed that in only 3 % of cases of CS the temporomandibular joint. This is a condition its characterized by being a metaplastic synovial connective tissue that manifests itself with the formation of small and multiple cartridges that detach, calcify and form free bodies within the joint space. We present the case of a 55-year-old woman with synovial chondromatosis of the temporomandibular joint, treated for 3 years under the diagnosis of temporomandibular disorders. Despite being considered a benign lesion, this can become locally aggressive, extending as in our report to the cranial fossa, thinning part of the temporal bone.

Synaptotagmin-1 overexpression under inflammatory conditions affects secretion in salivary glands from Sjögren's syndrome patients.

Sjögren's syndrome (SS) is an autoimmune exocrinopathy associated with severe secretory alterations by disruption of the glandular architecture integrity, which is fundamental for a correct function and localization of the secretory machinery. Syt-1, PI(4,5)P2 and Ca2+ are significant factors controlling exocytosis in different secretory cells, the Ca2+ role being the most studied. Salivary acinar cells from SS-patients show a defective agonist-regulated intracellular Ca2+ release together with a decreased IP3R expression level, and this condition may explain a reduced water release. However, there are not reports where Syt-1, PI(4,5)P2 and Ca2+ in acinar cells of SS patients had been studied. In the present study, we analyzed the expression and/or localization of Syt-1 and PI(4,5)P2 in acinar cells of labial salivary gland biopsies from SS-patients and control individuals. Also, we evaluated whether the overexpression of Syt-1 and the loss of cell polarity induced by TNF-α or loss of interaction between acinar cell and basal lamina, alters directionality of the exocytosis process, Ca2+ signaling and α-amylase secretion in a 3D-acini model stimulated with cholinergic or ß-adrenergic agonists. In addition, the correlation between Syt-1 protein levels and clinical parameters was evaluated. The results showed an increase of Syt-1 mRNA and protein levels, and a high number of co-localization points of Syt-1/STX4 and PI(4,5)P2/Ezrin in the acinar basolateral region of LSG from SS-patients. With regard to 3D-acini, Syt-1 overexpression increased exocytosis in the apical pole compared to control acini. TNF-α stimulation increased exocytic events in the basal pole, which was further enhanced by Syt-1 overexpression. Additionally, altered acinar cell polarity affected Ca2+ signaling and amylase secretion. Overexpression of Syt-1 was associated with salivary gland alterations revealing that the secretory dysfunction in SS-patients is linked to altered expression and/or localization of secretory machinery components together with impaired epithelial cell polarity. These findings provide a novel insight on the pathological mechanism implicated in ectopic secretory products to the extracellular matrix of LSG from SS-patients, which might initiate inflammation.

Colgajos Microvascularizados en Reconstrucción Maxilofacial: Avances de la Microcirugía / Microvascularized flaps in Maxillofacial Reconstruction: Advances in Microsurgery

RESUMEN: La reconstrucción de la cabeza y el cuello contempla avances importantes a lo largo de los años. Los colgajos microvasculares se han convertido en la primera opción de tratamiento en grandes defectos del territorio maxilofacial, mientras tanto, la tecnología con el uso de microscopía y luego las imágenes como CT, angiografía por tomografía computarizada, dispositivo ultrasónico, RNM o Doppler contribuyen a lograr una predictibilidad excepcional de estos colgajos microvasculares. Por lo general, la técnica de anastomosis consiste en una sutura de 9-0 en 360°, pero existen autores que han descrito diversos métodos que no son de sutura con un rendimiento aceptable. Existe un buen número de diferentes colgajos microvasculares, cuatro de ellos son los más comunes en la reconstrucción maxilofacial: fíbula, ilíaco, antebrazo radial, escápula. Además el colgajo anterolateral, muy útil en defectos de piel y tejidos blandos. La evolución de los colgajos microvasculares implica los colgajos quiméricos, muy útiles en defectos grandes. El objetivo de este artículo es describir y exponer el desarrollo de la microcirugía y las diversas opciones de colgajos microvasculares en la reconstrucción maxilofacial.

ABSTRACT: Head and neck reconstruction have shown important advances over the years. Microvasculars flaps transfer has become the first treatment option in large defects of the maxillofacial area. Meanwhile technology through the use of microscopy and the subsequent use of images such as CT, CT angiography, RNM or Doppler ultrasonic device, and additional new techniques have contributed to an exceptional predictability of these microvascular flaps. Typically, the anastomosis technique consists in 9-0 suture in 360°, but since the vascular flaps exist, authors have described diverse non-suture methods with acceptable performance. There are a number of different microvasculars flaps, four of them are the most common in maxillofacial reconstruction: fibula, iliac, radial forearm, scapula. In addition the anterolateral tight flap, very useful in skin and soft tissues defects. The microvascular flaps evolution involves the chimeric flaps that are useful in large defects. The aim of this article is to describe and expose microsurgery development and the diverse microvascular flap options in maxillofacial reconstruction.

Endoplasmic reticulum stress in autoimmune diseases: Can altered protein quality control and/or unfolded protein response contribute to autoimmunity? A critical review on Sjögren's syndrome.

For many years, researchers in the field of autoimmunity have focused on the role of the immune components in the etiopathogenesis of autoimmune diseases. However, some studies have demonstrated the importance of target tissues in their pathogenesis and the breach of immune tolerance. The immune system as well as target tissue cells (plasmatic, ß-pancreatic, fibroblast-like synoviocytes, thyroid follicular and epithelial cells of the lachrymal glands, salivary glands, intestine, bronchioles and renal tubules) share the characteristic of secretory cells with an extended endoplasmic reticulum (ER). The function of these cells depends considerably on a normal ER function and calcium homeostasis, so they can produce and secrete their main components, which include glycoproteins involved in antigenic presentation such as major histocompatibility complex (MHC) class I and II. All these proteins are synthesized and modified in the ER, and for this reason disturbances in the normal functions of this organelle such as protein folding, protein quality control, calcium homeostasis and redox balance, promote accumulation of unfolded or misfolded proteins, a condition known as ER stress. Autoimmune diseases are characterized by inflammation, which has been associated with an ER stress condition. Interestingly, patients with these diseases contain circulating auto-antibodies against chaperone proteins (such as Calnexin and GRP94), thus affecting the folding and assembly of MHC class I and II glycoproteins and their loading with peptide. The main purpose of this article is to review the involvement of the protein quality control and unfolded protein response (UPR) in the ER protein homeostasis (proteostasis) and their alterations in autoimmune diseases. In addition, we describe the interaction between ER stress and inflammation and evidences are shown of how autoimmune diseases are associated with an ER stress condition, with a special emphasis on the second most prevalent autoimmune rheumatic disease, Sjögren's syndrome.