Dynamically guided transantral piezoelectric endodontic microsurgery: A case report with technical considerations.

AIM: Endodontic microsurgery (EMS) of maxillary molars may represent a complex challenge to the clinician due to the location of the roots and the proximity of the maxillary sinus floor. This report aimed to describe the simultaneous use of a computer-assisted dynamic navigation (C-ADN) system and piezoelectric bony-window osteotomy for the transantral microsurgical approach of a maxillary left first molar with adequate root canal filling and symptomatic apical periodontitis. SUMMARY: This case report highlights the importance of C-ADN to carry out a minimally invasive buccal surgical access to palatal roots affected by apical periodontitis and provides a practical example to help clinicians make treatment decisions based on the available evidence. Clinical and tomographic evaluations were performed before the surgical procedure and at 24-month follow-up. This case was treated using a C-ADN system fitted to a piezotome for the buccal approach of the buccal roots, maxillary sinus membrane lifting, and for transantral location, root-end resection, cavity preparation, and filling of the palatal root. The navigation system allowed to achieve an accurate apical canal terminus location and root-end filling of the three roots with a minimally invasive piezoelectric crypt approach. At the 24-month follow-up examination, the patient remains asymptomatic, with normal periapical structures, and regeneration of maxillary sinus walls. It was concluded that the combination of dynamic navigation with piezoelectric bony-window osteotomy offers enhanced accuracy, tissue preservation, diminished risk of iatrogenic complications, and could maximize success and survival rates in transantral EMS.

DNA Methylation Effects on Van der Woude Syndrome Phenotypic Variability.

OBJECTIVE: Van der Woude Syndrome (VWS) presents with combinations of lip pits (LP) and cleft lip and/or cleft palate (CL/P, CPO). VWS phenotypic heterogeneity even amongst relatives, suggests that epigenetic factors may act as modifiers. IRF6, causal for 70% of VWS cases, and TP63 interact in a regulatory loop coordinating epithelial proliferation and differentiation in palatogenesis. We hypothesize that differential DNA methylation within IRF6 and TP63 regulatory regions underlie VWS phenotypic discordance. METHODS: DNA methylation of CpG sites in IRF6 and TP63 promoters and in an IRF6 enhancer element was compared amongst blood or saliva DNA samples of 78 unrelated cases. Analyses were done separately for blood and saliva, within each sex and in combination, and to address cleft type (CL/P ± LP vs. CPO ± LP) and phenotypic severity (any cleft + LP vs. any cleft only). RESULTS: For cleft type, blood samples showed higher IRF6 and TP63 promoter methylation on males with CPO ± LP compared to CL/P ± LP and on individuals with CPO ± LP compared to those with CL/P ± LP, respectively. Saliva samples showed higher IRF6 enhancer methylation on individuals with CPO ± LP compared to CL/P ± LP and contrary to above, lower TP63 promoter methylation on CPO ± LP compared to CL/P ± LP. For phenotypic severity, blood samples showed no differences; however, saliva samples showed higher IRF6 promoter methylation in individuals with any cleft + LP compared to those without lip pits. CONCLUSION: We observed differential methylation in IRF6 and TP63 regulatory regions associated with cleft type and phenotypic severity, indicating that epigenetic changes in IRF6 and TP63 can contribute to phenotypic heterogeneity in VWS.

The current and future costs of colorectal cancer attributable to red and processed meat consumption in Brazil.

BACKGROUND: Compelling evidence supports the association between red and processed meat consumption and increased risk of colorectal cancer. Herein, we estimated the current (2018) and future (2030) federal direct healthcare costs of colorectal cancer in the Brazilian Unified Health System attributable to red and processed meat consumption. Considering reduced red and processed meat consumption, we also projected attributable costs of colorectal cancer in 2040. METHODS: We retrieved information on red and processed meat consumption from two nationally representative dietary surveys, the Household Budget Survey 2008-2009 and 2017-2018; relative risks for colorectal cancer from a meta-analysis; direct healthcare costs of inpatient and outpatient procedures in adults ≥ 30 years with colorectal cancer (C18-C20) from 2008-2019 by sex. RESULTS: Attributable costs of colorectal cancer were calculated via comparative risk assessment, assuming a 10-year lag. In 2018, US$ 20.6 million (8.4%) of direct healthcare costs of colorectal cancer were attributable to red and processed meat consumption. In 2030, attributable costs will increase to US$ 86.6 million (19.3%). Counterfactual scenarios of reducing red and processed meat consumption in 2030 suggested that US$ 2.2 to 11.9 million and US$ 13 to 74 million could be saved in 2040, respectively. CONCLUSION: Red and processed meat consumption has an escalating economic impact on the Brazilian Unified Health System. Our findings support interventions and policies focused on primary prevention and cancer.

The future costs of cancer attributable to excess body weight in Brazil, 2030-2040.

BACKGROUND: Excess body weight (EBW), herein defined as body mass index (BMI) ≥25 kg/m2, is a well-known modifiable risk factor for cancer and a pivotal vector for growing healthcare costs. We estimated the future (2030) federal direct healthcare costs of cancer in the Brazilian Unified Health System (SUS) attributable to EBW. We also projected direct healthcare costs of cancer that could be potentially saved in 2040, considering counterfactual (alternative) scenarios of population-wide reductions in the BMI to be achievedin 2030. METHODS: We developed a macrosimulation model by sex using self-reported BMI data in adults ≥ 20 years who relied exclusively on the public health system from the Brazilian National Health Survey (PNS) 2019; relative risks for 12 types of cancer from the World Cancer Research Fund/American Institute Cancer Research (WCRF/AICR) meta-analysis; and nationwide registries of federal direct healthcare costs of inpatient and outpatient procedures in adults ≥30 years with cancer from 2008-2019. We calculated the attributable costs of cancer via comparative risk assessment, assuming a 10-year lag between exposure and outcome. We used the potential impact fraction (PIF) equation and the Monte Carlo simulation method to estimate the attributable costs and 95% uncertainty intervals, considering the theoretical-minimum-risk exposure and other counterfactual (alternative) scenarios of the EBW prevalence. We assessed the cancer costs attributable to EBW, multiplying PIF by the direct healthcare costs of cancer. RESULTS: In 2030, 2.4% or US$ 62.8 million in direct healthcare costs of cancer may be attributable to EBW. We projected potential savings of approximately US$ 10.3 to 26.6 million in 2040 by reducing the prevalence of EBW in 2030. CONCLUSIONS: We estimated high future costs of cancer attributable to EBW in Brazil. Our findings may support interventions and policies focused on the primary prevention of EBW and cancer.

Effect of chitosan solutions with or without fluoride on the protection against dentin erosion in vitro.

The aim of this study was to assess the protective effect of experimental solutions containing chitosan at different viscosities with or without fluoride (TiF4 /NaF) on dentin loss in vitro. Bovine dentin samples (n = 15) were prepared and allocated to one of the following treatments: (i) 0.5% chitosan (500 mPas); (ii) 0.5% chitosan (2,000 mPas); (iii) 0.042% NaF and 0.049% TiF4 ; (iv) as (iii) with addition of 0.5% chitosan (500 mPas); (v) as (iii) with addition of 0.5% chitosan (2,000 mPas); (vi) commercial solution with SnCl2 /AmF/NaF (positive control); or (vii) deionized water (negative control). The samples were submitted to pH cycling for 7 d (0.1% citric acid, 4 × 90 s d-1 ). The treatment was applied once a day for 30 s. The dentin loss was quantified using a contact profilometer. Three samples per group were evaluated using scanning electron microscopy. The dentin loss (µm) was submitted to anova and Tukey's test for differences between treatments. Among the treatments tested, only chitosan 500 mPas was able to statistically significantly reduce the dentin loss compared to the negative control, being similar to the positive control. TiF4 /NaF, whether with or without chitosan, had no protective effect. Chitosan 500 mPas and SnCl2 /AmF/NaF solutions have comparable protective effect against dentin erosion in vitro.

Antiviral Potential of Spondias mombin L. Leaves Extract Against Herpes Simplex Virus Type-1 Replication Using In Vitro and In Silico Approaches.

Spondias mobin leaves have been traditionally used for treating cold sores. The study investigated the mechanism of antiherpes action of S. mombin extract, fractions, and geraniin. Different concentrations of samples were used to evaluate the in vitro antiherpes activity (anti-HSV-1) in virucidal, post-infection, attachment, and penetration assays. The mechanism of action of geraniin was investigated considering the glycoproteins gB and gD of HSV-1 surface as potential molecular targets. Molecular docking simulations were carried out for both in order to determine the possible binding mode position of geraniin at the activity sites. The binding mode position was posteriorly optimized considering the flexibility of the glycoproteins. The chemical analysis of samples was performed by LC-MS and revealed the presence of 22 substances, which are hydrolysable tannins, O-glycosylated flavonoids, phenolic acids, and a carbohydrate. The extract, tannin-rich fraction and geraniin showed important in vitro virucidal activity through blocking viral attachment but showed no relevant inhibition of viral penetration. The in silico approaches demonstrated a high number of potential strong intermolecular interactions as hydrogen bonds between geraniin and the activity site of the glycoproteins, particularly the glycoprotein gB. In silico experiments indicated that geraniin is at least partially responsible for the anti-herpes activity through interaction with the viral surface glycoprotein gB, which is responsible for viral adsorption. These results highlight the therapeutic potential of S. mombin anti-herpes treatment and provides support for its traditional purposes. However, further studies are required to validate the antiviral activities in vivo, as well as efficacy in humans.

Tailoring structural properties of spray-dried methotrexate-loaded poly (lactic acid)/poloxamer microparticle blends.

Drug delivery systems can overcome cancer drug resistance, improving the efficacy of chemotherapy agents. Poly (lactic acid) (PLA) microparticles are an interesting alternative because their hydrophobic surface and small particle size could facilitate interactions with cells. In this study, two poloxamers (PLX 407 and 188) were applied to modulate the structural features, the drug release behavior and the cell viability from spray-dried microparticles. Five formulations with different PLA: PLX blend ratio (100:0, 75:25, 50:50, 25:50, and 0:100) were well-characterized by SEM, particle size analysis, FTIR spectroscopy, differential scanning calorimetry (DSC), and X-ray diffraction analysis (XRD). The spray-dried microparticles showed higher drug loading, spherical-shape, and smaller particle size. The type of poloxamer and blend ratio affected their structural and functional properties such as morphology, crystallinity, blend miscibility, drug release rate, and cell viability. The methotrexate (MTX), a model drug, was loaded in amorphous spray-dried microparticles. Moreover, the drug release studies demonstrated that PLX induced a leaching-effect of MTX from PLA: PLX blends, suggesting the formation of MTX/PLX micelles in aqueous medium. This finding was better established by cell viability assays. Therefore, biocompatible PLA: PLX blends showed promising in vitro results, and further in vivo studies will be performed to evaluate the performance of this chemotherapeutic agent.

Human Herpesvirus 8 in Perinatally HIV-infected Children with Interstitial Lung Disease.

INTRODUCTION: Human herpesvirus 8 (HHV-8) is associated with the pathogenesis of Kaposi Sarcoma and interstitial pneumonitis in adults. This study aims to evaluate association between HHV-8 and interstitial lung disease in HIV-infected children. METHODS: HIV-infected children with interstitial pneumonitis underwent lung biopsies in a tertiary hospital and were investigated for HHV-8, Epstein-Barr virus (EBV) and cytomegalovirus (CMV) using polymerase chain reaction (PCR) and immunohistochemistry in lung tissue. Peripheral blood PCR was also performed for HHV-8. RESULTS: From six patients included, PCR for HHV-8 was positive in lung samples in four children and in peripheral blood in one. PCR for EBV and CMV and immunohistochemical study for HHV-8, EBV and CMV in lung were negative in all patients. CONCLUSION: No previous cases of HHV-8-associated interstitial pneumonitis was described in HIV-infected children. An immunological disorder and an infectious agent might influence development of the lymphoid interstitial pneumonitis. HHV-8 may be this infectious trigger.

Nystatin and lidocaine pastilles for the local treatment of oral mucositis.

Oral mucositis (OM) is a common adverse reaction to radiotherapy and chemotherapy in oncology. Its treatment requires oral formulations that enhance therapy compliance, improve administration and ensure drug effectiveness. Solid dosage forms that act by slow dissolution, such as pastilles, are an effective alternative to mouthwashes, for their versatility, ease of administration and extended residence time in the oral cavity. The present work describes the development and stability studies of an innovative formulation of nystatin and lidocaine pastilles for the treatment of oral mucositis. Full pharmaceutical quality testing was carried out, including disintegration and dissolution testing, texture profile analysis, grittiness and an antifungal activity testing. A soft pastille formulation containing 0.25% lidocaine and 78,000 IU nystatin was obtained, presenting suitable pharmaceutical characteristics, as a disintegration time of 17 ± 2 min, dissolution rate and microbiological and physicochemical for 30 days when stored at 2-8 °C under light protection. Palatability was also evaluated, being well accepted by a panel of 38 healthy volunteers. This formulation allows an accurate drug dosing by the prescriber, while enabling the patients to control the retention time of the drugs in the oral cavity and consequently manage their pain treatment.

Characterization of Immunoglobulin A/G Responses During 3 Doses of the Human Papillomavirus-16/18 ASO4-Adjuvanted Vaccine.

BACKGROUND: Individuals receiving the human papillomavirus (HPV) vaccine develop high levels of circulating neutralizing antibodies. However, data about antibody responses in the cervix are limited. METHODS: This study was designed to describe the course of IgA/IgG responses in cervical secretions and in serum after intramuscular administration of the HPV16/18 AS04-adjuvant vaccine. An enzyme-linked immunosorbent assay for detection of IgA and IgG anti-HPV-VLP was developed for this purpose. RESULTS: Immunoglobulin G seroconversion after the second dose was observed in 100% of the participants and remained 1 month after the third dose. Regarding IgG reactivity in cervical secretions, conversion was observed in 85% of women after the final dose. Immunoglobulin A seroconversion was observed in 76.7% of women after the third dose. Lower levels of IgA were detected in the cervical mucus (28.3%) and decreased to 23.3% after the last dose. Comparing local and systemic IgG responses, positivity in both serum and cervical samples was observed in 85%, whereas in 15% only, the serum was IgG antibody positive. A weak agreement between local and systemic IgA responses was observed. Only 18.3% of participants were local and systemic IgA positive, 58.4% were positive only in serum, 5% were positive only in the cervix, and 18.3% were both local and systemic IgA antibody negative. CONCLUSIONS: After the third vaccination, there is a strong agreement between cervical and systemic IgG antibody responses and a weak agreement between cervical and systemic IgA antibody responses. The induction of IgA antibodies seems to be secondary to that of IgG antibodies in response to HPV intramuscular vaccination.

Phase Transitions of Isotropic to Anisotropic Biocompatible Lipid-Based Drug Delivery Systems Overcoming Insoluble Benznidazole Loading.

Previous studies reported low benznidazole (BNZ) loading in conventional emulsions due to the weak interaction of the drug with the most common oils used to produce foods or pharmaceuticals. In this study, we focused on how the type of surfactant, surfactant-to-oil ratio w/w (SOR) and oil-to-water ratio w/w (OWR) change the phase behavior of different lipid-based drug delivery systems (LBDDS) produced by emulsion phase inversion. The surfactant mixture composed of soy phosphatidylcholine and sodium oleate (1:7, w/w, hydrophilic lipophilic balance = 16) stabilized medium chain triglyceride in water. Ten formulations with the clear aspect or less turbid dispersions (five with the SOR ranging from 0.5 to 2.5 and five with the OWR from 0.06 to 0.4) were selected from the phase behavior diagram to assess structural features and drug-loading capacity. The rise in the SOR induced the formation of distinct lipid-based drug delivery systems (nanoemulsions and liquid crystal lamellar type) that were identified using rheological measurements and cross-polarized light microscopy images. Clear dispersions of small and narrow droplet-sized liquid-like nanoemulsions, Newtonian flow-type, were produced at SOR from 0.5 to 1.5 and OWR from 0.12 to 0.4, while clear liquid or gel-like liquid crystals were produced at SOR from 1.5 to 2.5. The BNZ loading was improved according to the composition and type of LBDDS produced, suggesting possible drug location among surfactant layers. The cell viability assays proved the biocompatibility for all of the prepared nanoemulsions at SOR less than 1.5 and liquid crystals at SOR less than 2.5, demonstrating their promising features for the oral or parenteral colloidal delivery systems containing benznidazole for Chagas disease treatment.

Effect of low-level laser therapy on bone repair: a randomized controlled experimental study.

The aim of this study was to investigate the effect of low-level laser therapy (LLLT) on bone repair in femoral fractures. Sixty adult Wistar rats were randomly assigned into one of two groups: group A (ostectomy + LLLT) or group B (ostectomy + sham laser). An experimental model of complete bone fracture was surgically created by removing a 2-mm fragment from the middle third of the femoral shaft. Data were analyzed on days 8, 13, and 18 after the fracture (subgroups 1, 2, and 3). Samples were assessed for changes in inflammatory infiltration; trabecular bone matrix, periosteal, and new bone formations; and changes in the expression of particular osteogenic-related proteins (osteocalcin, osteopontin, and osteonectin). Microscopic analysis revealed a significant decrease in inflammatory infiltration, intense trabecular bone matrix and periosteal formation, and an increase in newly formed bone after laser irradiation. We also found an increase in the expression of bone matrix proteins with LLLT, with a significant difference measured for osteocalcin in the LLLT group at day 8 (p = 0.007). We show that LLLT plays an important role in augmenting bone tissue formation, which is relevant to fracture healing. LLLT may therefore be indicated as an adjunct therapeutic tool in clinical practice for the treatment or recovery of nonunion injuries.

Chloroquine interferes with dengue-2 virus replication in U937 cells.

The objective of this study was to investigate the use of chloroquine (CLQ) as an antiviral agent against dengue. Chloroquine, an amine acidotropic drug known to affect intracellular exocytic pathways by increasing endosomal pH, was used in the in vitro treatment of U937 cells infected with dengue virus type 2 (DENV-2). Viral replication was assessed by quantification of virus produced through detection of copy numbers of DENV-2 RNA, plaque assay and indirect immunofluorescence. qRT-PCR and plaque assays were used to quantify the DENV-2 load in infected U937 cells after CLQ treatment. It was found that a dose of 50 µg/mL of CLQ was not toxic to the cells and resulted in significantly less virus production in infected U937 cells than occurred in untreated cells. In the present work, CLQ was effective against DENV-2 replication in U937 cells, and also caused a statistically significant reduction in expression of proinflammatory cytokines. The present study indicates that CLQ may be used to reduce viral yield in U937 cells.

Pain Neuroscience Education in elective surgery patients: study protocol for a randomised controlled trial.

BACKGROUND: Pain Neuroscience Education (PNE) consists of an educational strategy that seeks to understand the biological processes of pain and how to control it. The main objective of this study will be to evaluate the impact of PNE on outcomes related to the postoperative period. The hypothesis is that the intervention may positively influence postoperative recovery, contributing to pain control, clinical indications, acceptance and consumption of analgesics and other pharmacological drugs that contribute to its control, as well as psychological aspects, such as anxiety, depression and pain catastrophising. METHODS AND ANALYSIS: This will be an open, parallel, multicentre and randomised controlled clinical trial. A total of 100 participants aged between 18 and 59 years of age, of both genders, who are going to have elective general surgery will be evaluated. The intervention group will participate in a preoperative pain neuroscience educational session and also receive usual preoperative care, while the control group receives usual preoperative care as well. The educational session will last 30 min and consists of a video (5:20 min), a questionnaire about the content, time for participants to express their beliefs, thoughts and doubts. Participants will be evaluated preoperatively and there will be one postintervention evaluation. The intensity and characteristics of pain and anxiety are evaluated as primary outcomes. As secondary outcomes, pain catastrophising and depression are taken into account. ETHICS AND DISSEMINATION: The project was approved by the Research Ethics Committee of the Faculty of Ceilandia, the Research Ethics Committee of the Institute of Strategic Health Management of the Federal District and the Research Council of the Hospital of Brasília-Rede Dasa (CAAE: 28572420.3.0000.8093). Recruitment began in June of 2023. All participants were included in the study only after their written consent. All data obtained will be analysed and distributed through publication in journals and at scientific events. TRIAL REGISTRATION NUMBER: Brazilian Registry of Clinical Trials (ReBEC) (RBR-23mr7yy).

Evaluation of microhardness of short fiber-reinforced composites inside the root canal after different light curing methods - An in vitro study.

OBJECTIVES: Short fiber-reinforced composite (SFRC) materials make it possible to reinforce root canal treated teeth with individualized, directly layered intraradicular posts (the Bioblock technique). The question arises, however, as to whether the photopolymerization of the material is sufficient deep within the root canal space and if it can be improved through different light-conducting options. Our study aimed to investigate the hardness of intraradicular SFRC material applied using the Bioblock technique and cured with various illumination methods, as measured through nanoindentation. MATERIALS AND METHODS: For this investigation, thirty plastic artificial teeth that had undergone root canal treatment were selected. These teeth were randomly divided into six study groups (Group 1-6; each group consisting of 5 teeth). The restoration procedures involved the use of SFRC or conventional composite materials, placed 6 mm apically from the root canal orifice. In Group 1 and 2, a conventional composite was used, whereas in Group 3-6, SFRC was employed for interradicular reinforcement (with a layered technique in Group 3 and 4 and a bulk-fill technique in Group 5 and 6). A modified light source was utilized for photopolymerization in Group 2, 4, and 6, whereas in Group 3 and 5, the polymerization light was directed through a prefabricated glass fiber posts. The control group (Group 1) utilized conventional composite material with a standard light-curing method. Following embedding and sectioning, the hardness of the composite materials was measured at 2 mm intervals within the root canal (1st, 2nd, 3rd measurements, in the coronal to apical direction). RESULTS: During the 1st measurement, light curing conducted through the glass fiber posts (Group 3 and 5) led to markedly higher hardness levels compared to the groups restored with conventional composite (control group with p = 0.002, p = 0.001, and Group 2 with p = 0.043, p = 0.034, respectively). In the 2nd measurement, only Group 5 demonstrated significantly greater hardness in comparison to the control group (p = 0.003) and Group 2 (p = 0.015). However, in the 3rd measurement, no statistically significant differences were observed among the groups. CONCLUSION: light curing through the glass fiber post provides outstanding hardness for the SFRC material in the apical layer in the root canal.

Pretreatment HIV-1 Resistance in Argentina: Results from the Second Surveillance Study Following World Health Organization Guidelines (2019).

More than 62,000 individuals are currently on antiretroviral treatment within the public health system in Argentina. In 2019, more than 50% of people on ART received non-nucleoside reverse transcriptase inhibitors (NNRTIs). In this context, the second nationwide HIV-1 pretreatment drug resistance surveillance study was carried out between April and December 2019 to assess the prevalence of HIV-1 drug resistance in Argentina using the World Health Organization guidelines. This was a nationwide cross-sectional study enrolling consecutive 18-year-old and older individuals starting ARVs at 19 ART-dispensing centers. This allowed us to estimate a point prevalence rate of resistance-associated mutations (RAMs) with a confidence interval (CI) of 5% (for the total population and for those without antiretroviral exposure). Four-hundred forty-seven individuals were included in the study. The prevalence of mutations associated with resistance was detected in 27.7% (95% CI 25.6-34.9%) of the population. For NNRTI, it was 19.6% (95% CI 16.3-24.5%), for integrase strand transfer inhibitor (INSTI) 6.1% (95% CI 6.1-11.9%), for nucleoside/nucleotide reverse transcriptase inhibitor (NRTI) 3% (95% CI 1.9-5.9%), and for protease inhibitors 1.5% (95% CI 0.7-3.6%). Naive individuals had variants of resistance to NRTIs in 16.8% (95% CI 12.8-21.4) and 5.7% (95% CI 2.9-15.9) to INSTI. For experienced individuals, the prevalence of variants associated with resistance was 30.38% (95% CI 20.8-42.2) for NRTIs and 7.7% (95% CI 2.9-15.9) for INSTI. This study shows an increase in the frequency of nonpolymorphic RAMs associated with resistance to NNRTI. This study generates the framework of evidence that supports the use of schemes based on high genetic barrier integrase inhibitors as the first line of treatment and the need for the use of resistance test before prescribing schemes based on NNRTI. We report for the first time the presence of a natural polymorphism associated with the most prevalent recombinant viral form in Argentina and the presence of a mutation linked to first-line integrase inhibitors such as raltegravir.

Retrospective Follow-up Assessment of Risk Variables Influencing the Outcome of Autologous Tooth Transplantation.

INTRODUCTION: This retrospective study aimed to identify which patient-, donor tooth-, recipient site-, and surgical procedure-related variables may influence the outcome of tooth autotransplantation. METHODS: The sample included 128 autotransplants performed in 122 patients. Single-visit clinical/imaging examinations were used to define the outcome as successful, survival, or failure. The association of potential indicators with the survival or failure categories was analyzed individually and adjusted for confounders through multivariate logistic regression models. RESULTS: After a follow-up period of 1 to 30.11 years, success was achieved in 71.8% of autotransplants, whereas the survival and failure groups had rates of 14.1% each, and the grouped success/survival rate reached 85.9%. An extraoral time >15 minutes and difficult handling/placement were strong/independent risk covariates for survival and failure categories (odds ratio >1, P < .05). Additionally, unerupted/partially erupted status of the donor tooth was a significant indicator for survival, whereas deficient bone level at the recipient site, surgical extraction, poor initial stability, and lack of prophylactic antibiotics were independently linked to failure (odds ratio > 1, P < .05). The root morphology and socket status acted as modifiers of the effect of the recipient site location on the survival group (P > .05). CONCLUSION: Based on the results of this study, unerupted/partially erupted status of the donor tooth, surgical extraction, total extraoral time >15 minutes, deficient recipient's bone level, difficult handling/placement of the autotransplant, poor initial stability, and lack of prophylactic antibiotics during the surgical procedure must be considered with caution when performing autotransplantation because of their deleterious influence on the outcome.

Whole genome sequencing of a family with autosomal dominant features within the oculoauriculovertebral spectrum.

Background: Oculoauriculovertebral Spectrum (OAVS) encompasses abnormalities on derivatives from the first and second pharyngeal arches including macrostomia, hemifacial microsomia, micrognathia, preauricular tags, ocular and vertebral anomalies. We present genetic findings on a three-generation family affected with macrostomia, preauricular tags and uni- or bilateral ptosis following an autosomal dominant pattern. Methods: We generated whole genome sequencing data for the proband, affected parent and unaffected paternal grandparent followed by Sanger sequencing on 23 family members for the top 10 candidate genes: KCND2, PDGFRA, CASP9, NCOA3, WNT10A, SIX1, MTF1, KDR/VEGFR2, LRRK1, and TRIM2 We performed parent and sibling-based transmission disequilibrium tests and burden analysis via a penalized linear mixed model, for segregation and mutation burden respectively. Next, via bioinformatic tools we predicted protein function, mutation pathogenicity and pathway enrichment to investigate the biological relevance of mutations identified. Results: Rare missense mutations in SIX1, KDR/VEGFR2, and PDGFRA showed the best segregation with the OAV phenotypes in this family. When considering any of the 3 OAVS phenotypes as an outcome, SIX1 had the strongest associations in parent-TDTs and sib-TDTs (p=0.025, p=0.052) (unadjusted p-values). Burden analysis identified SIX1 (RC=0.87) and PDGFRA (RC=0.98) strongly associated with OAVS severity. Using phenotype-specific outcomes, sib-TDTs identified SIX1 with uni- or bilateral ptosis (p=0.049) and ear tags (p=0.01), and PDGFRA and KDR/VEGFR2 with ear tags (both p<0.01). Conclusion: SIX1, PDGFRA, and KDR/VEGFR2 are strongly associated to OAVS phenotypes. SIX1 has been previously associated with OAVS ear malformations and is co-expressed with EYA1 during ear development. Efforts to strengthen the genotype-phenotype co-relation underlying the OAVS are key to discover etiology, family counseling and prevention.

Efficacy of sperm motility after processing and incubation to predict pregnancy after intrauterine insemination in normospermic individuals.

BACKGROUND: Intrauterine insemination (IUI) is widely used to treat infertility, and its adequate indication is important to obtain good pregnancy rates. To assess which couples could benefit from IUI, this study aimed to evaluate whether sperm motility using a discontinuous gradient of different densities and incubation in CO2 in normospermic individuals is able to predict pregnancy. METHODS: A total of 175 couples underwent 175 IUI cycles. The inclusion criteria for women were as follows: 35 years old or younger (age range: from 27 to 35 years) with normal fallopian tubes; endometriosis grades I-II; unexplained infertility; nonhyperandrogenic ovulatory dysfunction. Men with normal seminal parameters were also included. All patients underwent ovarian stimulation with clomiphene citrate and human hMG or r-FSH. When one or (at most) three follicles measuring 18 to 20 mm were observed, hCG (5000 UI) or r-hCG (250 mcg) was administered and IUI performed 36-40 h after hCG. Sperm processing was performed using a discontinuous concentration gradient. A 20 microliters aliquot was incubated for 24 h at 37 degrees C in 5% CO2 following a total progressive motility analysis. The Mann-Whitney and Chi-square tests, as well as a ROC curve were used to determine the cutoff value for motility. RESULTS: Of the 175 couples, 52 (in 52 IUI cycles) achieved clinical pregnancies (CP rate per cycle: 29.7%). The analysis of age, duration and causes of infertility did not indicate any statistical significance between pregnancy and no pregnancy groups, similar to the results for total sperm count and morphology analyses, excluding progressive motility (p < 0.0001). The comparison of progressive motility after processing and 24 h after incubation between these two groups indicated that progressive motility 24 h after incubation was higher in the pregnancy group. The analysis of the progressive motility of the pregnancy group after processing and 24 h after incubation has not shown any motility difference at 24 h after incubation; additionally, in couples who did not obtain pregnancy, there was a statistically significant decrease in progressive motility 24 h after incubation (p < 0.0001). The ROC curve analysis generated a cutoff value of 56.5% for progressive motility at 24 h after incubation and this cutoff value produced 96.1% sensitivity, 92.7% specificity, 84.7% positive predictive value and 98.3% negative predictive value. CONCLUSIONS: We concluded that the sperm motility of normospermic individuals 24 h after incubation at 37 degrees C in 5% CO2, with a cutoff value of 56.5%, is predictive of IUI success.

Chloroquine inhibits dengue virus type 2 replication in Vero cells but not in C6/36 cells.

Dengue viruses are the most important arthropod-borne viruses in terms of morbidity and mortality in the world. Since there is no dengue vaccine available for human use, we have set out to investigate the use of chloroquine as an antiviral drug against dengue. Chloroquine, an amine acidotropic drug known to affect intracellular exocytic pathways by increasing endosomal pH, was used in the in vitro treatment of Vero and C6/36 cells infected with dengue virus type 2 (DENV-2). Real-time RT-PCR and plaque assays were used to quantify the DENV-2 load in infected Vero and C6/36 cells after chloroquine treatment. Our results showed that a dose of 50 µg/ml of chloroquine was not toxic to the cells and induced a statistically significant inhibition of virus production in infected Vero cells when compared to untreated cells. In C6/36 cells, chloroquine does not induce a statistically significant difference in viral replication when compared to untreated cells, showing that this virus uses an unlikely pathway of penetration in these cells, and results were also confirmed by the plaque assay (PFU). These data suggest that the inhibition of virus infection induced by chloroquine is due to interference with acidic vesicles in mammalian cells.