Omicron BA.4/5 Neutralization and T-Cell Responses in Organ Transplant Recipients After Booster Messenger RNA Vaccine: A Multicenter Cohort Study.

BACKGROUND: In solid organ transplant (SOT) recipients, the primary vaccination series against Coronavirus Disease 2019 is 3 doses followed by boosters. We determined whether a fourth dose booster induced Omicron BA.4/5 neutralizing antibodies (nAbs) and T cells in a large multicenter cohort study. METHODS: Serum was collected 4-6 weeks post-third and post-fourth doses of messenger RNA vaccine in 222 SOT recipients. nAbs were measured using a pseudovirus neutralization assay that targeted the Omicron BA.4/5 spike protein. A subset underwent T-cell testing. RESULTS: The median age of the cohort was 63 years (interquartile range [IQR], 50-68) with 61.7% men. BA.4/5 nAb detection increased from 26.6% (59 of 222) post-third dose to 53.6% (119 of 222) post-fourth dose (P < .0001). In patients with breakthrough infection prior to the fourth dose (n = 27), nAbs were detected in 77.8% and median nAb titers were significantly higher compared with those with 4 vaccine doses alone (P < .0001). Factors associated with a low BA.4/5 neutralization response after the fourth dose were older age (odds ratio [OR], 0.96; 95% confidence interval [CI], .94-.99), mycophenolate use (OR, 0.39; 95% CI, .20-.77) and prednisone use (OR, 0.34; 95% CI, .18-.63), and vaccine type (OR, 0.72; 95% CI, .51-.99), while breakthrough infection prior to the fourth dose (OR, 3.6; 95% CI, 1.3-9.9) was associated with a greater nAb response. Polyfunctional BA.4/5-specific CD4+ T cells significantly increased after 4 doses and were identified in 76.9% of patients at a median frequency of 213/106 cells (IQR, 98-650). CONCLUSIONS: In summary, a booster significantly increases BA.4/5-specific neutralization and polyfunctional CD4+ T-cell responses, suggesting protection from severe disease even with new Omicron variants. However, SOT recipients who are older and on mycophenolate and prednisone need additional preventative strategies.

Identifying social science engagement within agroecology: Classifying transdisciplinary literature with a semi-automated textual classification method.

Interdisciplinary and transdisciplinary fields of inquiry and action have been important academic frontiers in recent years. The field of agroecology is a prime example of transdisciplinarity. With roots in the biophysical sciences, social sciences, and peasant movements, publications in agroecology have been growing rapidly in recent decades. Here we explain a method-the script-expert adaptive classification (SEAC) method-that allows us to examine the engagements between agroecology and the social sciences by identifying publications within the agroecological literature that engage with social science at various levels. Using the term "agroecology" and its iterations, we gathered a corpus of agroecology literature up to and including 2019 with 12,398 unique publications from five publication databases-Scopus, Web of Science, Agricola, CAB Direct, and EconLit. Using the SEAC method we then classified each publication as engaged, partially engaged, and not engaged with social sciences and separated this Agroecology Corpus 2019 into three corpora: agroecology engaged with social sciences (with 3,125 publications), agroecology not engaged with social sciences (with 7,039 publications), and agroecology with uncertain engagement with social science (with 2,234 publications) or unclassifiable. This article explains the SEAC method in detail so other transdisciplinary scholars can replicate and/or adapt it for similar purposes. We also assess the SEAC method's value in identifying social science publications relative to the classification systems of the major multidisciplinary bibliographic databases, Scopus, and Web of Science. We conclude by discussing the strengths and weaknesses of the SEAC method and by pointing to further questions about agroecology and the social sciences to be asked of the corpora.

Single-cell mass cytometry on peripheral cells in Myasthenia Gravis identifies dysregulation of innate immune cells.

Myasthenia Gravis (MG) is a neurological autoimmune disease characterized by disabling muscle weaknesses due to anti-acetylcholine receptor (AChR) autoantibodies. To gain insight into immune dysregulation underlying early-onset AChR+ MG, we performed an in-depth analysis of peripheral mononuclear blood cells (PBMCs) using mass cytometry. PBMCs from 24 AChR+ MG patients without thymoma and 16 controls were stained with a panel of 37 antibodies. Using both unsupervised and supervised approaches, we observed a decrease in monocytes, for all subpopulations: classical, intermediate, and non-classical monocytes. In contrast, an increase in innate lymphoid cells 2 (ILC2s) and CD27- γδ T cells was observed. We further investigated the dysregulations affecting monocytes and γδ T cells in MG. We analyzed CD27- γδ T cells in PBMCs and thymic cells from AChR+ MG patients. We detected the increase in CD27- γδ T cells in thymic cells of MG patients suggesting that the inflammatory thymic environment might affect γδ T cell differentiation. To better understand changes that might affect monocytes, we analyzed RNA sequencing data from CD14+ PBMCs and showed a global decrease activity of monocytes in MG patients. Next, by flow cytometry, we especially confirmed the decrease affecting non-classical monocytes. In MG, as for other B-cell mediated autoimmune diseases, dysregulations are well known for adaptive immune cells, such as B and T cells. Here, using single-cell mass cytometry, we unraveled unexpected dysregulations for innate immune cells. If these cells are known to be crucial for host defense, our results demonstrated that they could also be involved in autoimmunity.

Farm resilience during the COVID-19 pandemic: The case of California direct market farmers.

CONTEXT: The COVID-19 pandemic caused significant shocks to U.S. food systems at multiple scales. While disturbances to long-distance supply chains received substantial attention in national media, local supply chains experienced mixed impacts. As broad closures of schools, restaurants, and other businesses sourcing from local farmers removed key marketing channels for many direct market farmers, consumer interest in Community Supported Agriculture (CSA), farmers markets, and on-farm and online direct farm sales increased. OBJECTIVE: In this paper, we examine the resilience and vulnerability of farmers during the March 2020 through December 2020 period of the COVID-19 pandemic. We focus on California farmers and ranchers engaged in direct market sales. METHODS: Through a widely disseminated survey, we collected responses from 364 farmers and used these data to answer the following questions about direct market farmers in California: 1) What were direct market farmers' experiences of the pandemic from March 2020 through December 2020? 2) Which factors (e.g., relationships, institutions, market channels) did farmers report enhanced their resilience during the pandemic? 3) Which individual and operational factors were significantly associated with resilience during the pandemic? And finally, 4) how do the farmer-reported factors compare to the statistically significant factors associated with resilience? We created three dependent variables-ability to respond to the pandemic, concern about pandemic impacts, and change in profitability-to operationalize several aspects of resilience and examine their association with individual and operational characteristics through a series of ordered logistic regression models. RESULTS AND CONCLUSIONS: Across both the quantitative models and the farmer reported factors, we found that farmers who increased their use of online sales and marketing during the first year of the pandemic, had larger-scale farms, and had more on-farm crop and livestock diversity were more resilient to the shocks of the pandemic. We also found that greater use of non-direct-to-consumer market channels was associated with less resilience. The characteristics of the farming operations played a relatively larger role in predicting resilience compared to the individual characteristics of the farmers surveyed. SIGNIFICANCE: This study gives a detailed picture of how California direct market farmers fared during the pandemic and the characteristics associated with greater resilience. As short and long-term disruptions become increasingly common in agriculture, policies and programs can leverage support to direct market farmers, particularly direct-to-consumer farmers, as a strategy to strengthen farmer resilience.

Efficacy of Botulinum Toxin Type-A I in the Improvement of Mandibular Motion and Muscle Sensibility in Myofascial Pain TMD Subjects: A Randomized Controlled Trial.

This study assessed the effects of botulinum toxin type A (BoNT-A) in mandibular range of motion and muscle tenderness to palpation in persistent myofascial pain (MFP) patients (ReBEC RBR-2d4vvv). Eighty consecutive female subjects with persistent MFP, were randomly divided into four groups (n = 20): three BoNT-A groups with different doses and a saline solution group (placebo control group). Treatments were injected bilaterally in the masseter and anterior temporalis muscle in a single session. Clinical measurements of mandibular movements included: pain-free opening, maximum unassisted and assisted opening, and right and left lateral excursions. Palpation tests were performed bilaterally in the masseter and temporalis muscle. Follow-up occurred 28 and 180 days after treatment. For the statistical analysis the Mann-Whitney U-test with Bonferroni correction was used for groups comparisons. Regardless of dose, all parameters of mandibular range of motion significantly improved after 180 days in all BoNT-A groups, compared with the control group. Palpation pain over the masseter and temporalis muscles were significantly reduced in all BoNT-A groups regardless of dose, compared with the control group, after 28 and 180 days of treatment. Independent of doses, BoNT-A improved mandibular range of motion and muscle tenderness to palpation in persistent MFP patients.

Evaluation of Three Anti-SARS-CoV-2 Serologic Immunoassays for Post-Vaccine Response.

BACKGROUND: In North America, both messenger RNA (mRNA) vaccines, Pfizer-BioNTech BNT162b2, and Moderna mRNA-1273, each utilizing a 2-dose regimen, have started to be administered to individuals. METHODS: We evaluated the quantitative serologic antibody response following administration of either a single dose or both doses of an mRNA severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine in a cohort of 98 participants (88 healthcare workers [HCW] and 10 solid organ transplant [SOT] recipients). Antibody levels were compared across 3 immunoassays: Elecsys Anti-SARS-CoV-2 S (Roche Diagnostics), SARS-CoV-2 TrimericS IgG (DiaSorin), and SARS-CoV-2 IgG II Quant (Abbott). RESULTS: Among HCW, sensitivity ranged from 100% (Roche), 99% (Abbott) and 98% (DiaSorin). The SARS-CoV-2 IgG II Quant and SARS-CoV-2 TrimericS IgG assays showed good agreement with a Pearson correlation coefficient of R = 0.95. Pearson correlation coefficients of R = 0.82 and 0.83 were obtained for Elecsys Anti-SARS-CoV-2 S vs SARS-CoV-2 TrimericS IgG and SARS-CoV-2 IgG II Quant vs Elecsys Anti-SARS-CoV-2 S, respectively. Significant differences in antibody levels between HCW and SOT recipients were observed. A decrease in antibody levels from time of vaccine administration to blood draw was evident. Among those with a second dose, an increase in antibody levels with increased time between administration of the first and second dose was observed. CONCLUSIONS: The absolute values generated from each of the assay platforms are not interchangeable. Antibody levels differed with increased time between vaccine administration and with increased time between administration of the first and second dose. Further, significant differences in antibody levels between HCW and SOT recipients were observed.

Lung transplant recipient attitudes and beliefs on accepting an organ that is positive for hepatitis C virus.

INTRODUCTION: Little is known about patient perceptions regarding HCV+ organ use in non-HCV-infected recipients. This study examined factors influencing the decision to accept HCV+ organs and the resulting impacts. METHODS: Adult lung transplant (LT) patients or candidates who had consented to receive an HCV+ organ completed a survey including multiple choice, a five-point Likert scale, and free-text answers. A total of 67 LT recipients or candidates who had consented to receive HCV+ organs were enrolled, of which 21/67 (31%) received HCV+ lungs, 39 (58%) HCV- lungs, and seven (10%) were still waiting. RESULTS: Pre-transplant, 50/67 (75%) patients felt it was either "completely safe" or "very safe" to accept an HCV+ organ. Although 22/67 (33%) said they never or rarely took risks, they still made the decision to accept an HCV+ organ. Common reported reasons were desperation, perception of having "no choice," and increasing symptom severity. In the subset of patients that were transplanted with an HCV+ organ (n = 21), only 12.5% reported second thoughts about accepting. Post-transplant, the majority (87.5%) never felt any anxiety about HCV and most (83%) reported no impact from HCV. Perception of treatment tolerability and ease was highly favorable. CONCLUSION: Use of HCV+ organs demonstrated minimal detrimental perceived impacts on lung transplant patients. Patients generally found the experience to be very positive.

Prospective observational study and serosurvey of SARS-CoV-2 infection in asymptomatic healthcare workers at a Canadian tertiary care center.

Health care workers (HCWs) are at higher risk for SARS-CoV-2 infection and may play a role in transmitting the infection to vulnerable patients and members of the community. This is particularly worrisome in the context of asymptomatic infection. We performed a cross-sectional study looking at asymptomatic SARS-CoV-2 infection in HCWs. We screened asymptomatic HCWs for SARS-CoV-2 via PCR. Complementary viral genome sequencing was performed on positive swab specimens. A seroprevalence analysis was also performed using multiple assays. Asymptomatic health care worker cohorts had a combined swab positivity rate of 29/5776 (0.50%, 95%CI 0.32-0.75) relative to a comparative cohort of symptomatic HCWs, where 54/1597 (3.4%) tested positive for SARS-CoV-2 (ratio of symptomatic to asymptomatic 6.8:1). SARS-CoV-2 seroprevalence among 996 asymptomatic HCWs with no prior known exposure to SARS-CoV-2 was 1.4-3.4%, depending on assay. A novel in-house Coronavirus protein microarray showed differing SARS-CoV-2 protein reactivities and helped define likely true positives vs. suspected false positives. Our study demonstrates the utility of routine screening of asymptomatic HCWs, which may help to identify a significant proportion of infections.

Efficacy and Safety of Botulinum Toxin Type A on Persistent Myofascial Pain: A Randomized Clinical Trial.

This study assessed the safety and efficacy of three different doses of BoNT-A for persistent myofascial pain (MFP). One hundred female subjects were randomly assigned into five groups (n = 20): oral appliance (OA), saline solution (SS) and three BoNT-A groups with different doses. Pain intensity and pressure pain threshold were evaluated up to 24 weeks after treatment. Adverse effects related to muscle contraction, masticatory performance, muscle thickness and mandibular bone volume were also assessed. Changes over time were compared within and between groups. The "nparLD" package and Wilcoxon signed-rank test were used to analyze the data. BoNT-A reduced pain intensity (p < 0.0001) and increased pressure pain threshold (p < 0.0001) for up to 24 weeks compared to the placebo. No differences were found between BoNT-A and OA at the last follow-up. A transient decline in masticatory performance (p < 0.05) and muscle contraction (p < 0.0001), and a decrease in muscle thickness (p < 0.05) and coronoid and condylar process bone volume (p < 0.05) were found as dose-related adverse effects of BoNT-A. Regardless of the dose, BoNT-A was as effective as OA on MFP. Notwithstanding, due to BoNT-A dose-related adverse effects, we suggest the use of low doses of BoNT-A in MFP patients that do not benefit from conservative treatments.

Attitudes of Liver Transplant Candidates Toward Organs From Increased-Risk Donors.

Increased-risk donor (IRD) organs make up a significant proportion of the deceased organ donor pool but may be declined by patients on the waiting list for various reasons. We conducted a survey of patients awaiting a liver transplant to determine the factors leading to the acceptance of an IRD organ as well as what strategies could increase the rate of acceptance. Adult liver transplant candidates who were outpatients completed a survey of 51 questions on a 5-point Likert scale with categories related to demographics, knowledge of IRDs, and likelihood of acceptance. A total of 150 transplant candidates completed the survey (age 19-80 years). Male patients constituted 67.3%. Many patients (58.7%) had postsecondary education. Only 23.3% of patients had a potential living donor, and 58/144 (40.3%) were not optimistic about receiving an organ in the next 3 months. The overall IRD organ acceptance rate was 41.1%, whereas 26.2% said they would decline an IRD organ. Women were more likely to accept an IRD organ (54.3% versus 34.7%; P = 0.02). Those who had a college education or higher tended to have lower IRD organ acceptability (28.3% versus 47.4%; P = 0.07). Acceptability also increased as the specified transmission risk of human immunodeficiency virus or hepatitis C virus decreased (P < 0.001). Patients were also more likely to accept an IRD organ if they were educated on the benefits of IRD organs (eg, knowledge that an IRD organ was of better quality increased overall acceptance from 41.1% to 63.3%; P < 0.001). Our survey provides insight into liver transplant candidates who would benefit from greater education on IRD organs. Strategies targeting specific educational points are likely to increase acceptability.

Functional lability of RNA-dependent RNA polymerases in animals.

RNA interference (RNAi) requires RNA-dependent RNA polymerases (RdRPs) in many eukaryotes, and RNAi amplification constitutes the only known function for eukaryotic RdRPs. Yet in animals, classical model organisms can elicit RNAi without possessing RdRPs, and only nematode RNAi was shown to require RdRPs. Here we show that RdRP genes are much more common in animals than previously thought, even in insects, where they had been assumed not to exist. RdRP genes were present in the ancestors of numerous clades, and they were subsequently lost at a high frequency. In order to probe the function of RdRPs in a deuterostome (the cephalochordate Branchiostoma lanceolatum), we performed high-throughput analyses of small RNAs from various Branchiostoma developmental stages. Our results show that Branchiostoma RdRPs do not appear to participate in RNAi: we did not detect any candidate small RNA population exhibiting classical siRNA length or sequence features. Our results show that RdRPs have been independently lost in dozens of animal clades, and even in a clade where they have been conserved (cephalochordates) their function in RNAi amplification is not preserved. Such a dramatic functional variability reveals an unexpected plasticity in RNA silencing pathways.

Toxina Botulínica para el Tratamiento de los Desórdenes Temporomandibulares / Botulinum Toxin for the Treatment of Temporomandibular Disorders

RESUMEN: Los Trastornos Temporomandibulares (TTM) consisten en un conjunto de condiciones patológicas que afectan la articulación temporomandibular (ATM), los músculos de la masticación y las estructuras asociadas. Estudios epidemiológicos estiman que 40 % a 75 % de la población adulta presenta por lo menos un signo de TTM, como ruido articular y 33 % por lo menos un síntoma, como dolor facial o en la ATM. La toxina botulínica (BoNT) es una neurotoxina producida por la bacteria anaerobia Clostridium botulinum, solo la toxina A y B son utilizadas en la práctica clínica después de la aprobación de la Food and Drug Administration en 1989 y 2000; actualmente continúan siendo utilizadas para tratar varias condiciones de dolor, incluyendo la espasticidad muscular, distonia, dolor de cabeza y dolor miofascial. Las propuestas del mecanismo de acción fueron sugeridas a mediados de 1950 manifestando que esta neurotoxina posee alta afinidad con la sinapsis colinérgicas, ocasionando un bloqueo en la liberación de acetilcolina de esos terminales nerviosos, sin alterar la conducción neural de las señales eléctricas o en la síntesis de almacenamiento de acetilcolina. La inyección intramuscular en dosis y localización apropiada, provoca denervación química parcial y disminución de la contractura, sin ocasionar parálisis completa a lo que se le atribuye ser un innovador y eficaz método de tratamiento para el dolor crónico asociada con hiperactividad de los músculos masticatorios. La toxina botulínica tipo A es ser una alternativa para el control de la sintomatología dolorosa presente en los TTMs de etiología miogénica. Los autores recomiendan realizar un correcto diagnóstico, ya que la indicación de este tipo de tratamiento con diagnósticos incorrectos lleva a resultados inciertos.

ABSTRACT: Temporomandibular Disorders (TMD) is a term that was used to describe a set of pathological conditions that affect temporomandibular joint (TMJ), muscles of mastication and associated structures. Epidemiological studies estimates that 40 % to 75 % of the adult population has at least one sign of TMD, such as joint noise and 33 % presented some symptom such as facial or joint pain. Botulinum toxin is a neurotoxin produced by the anaerobic bacterium Clostridium botulinum. There are two types of toxin (A and B) used in clinical practice that were approved by Food and Drug Administration in 1989 and 2000. These medications are in use to treat various diseases including muscle spasticity, dystonia, headache and myofascial pain. The mechanisms of action were suggested in the mid-1950s, this neurotoxin seems to have an action at cholinergic synapses, causing a block in the release of acetylcholine from the nerve terminals without altering the neural conduction of the electrical signals. The synthesis and storage of acetylcholine were preserved. Intramuscular injection in appropriate doses and location causes partial chemical denervation and decreased contracture, without causing complete paralysis of muscles. Due to this features, it has been considered an innovative and effective method to treat chronic pain associated with hyperactivity of masticatory muscles. The botulinum toxin type A appears to be an alternative method for pain control present in TMDs that have myogenic etiology. The authors recommend a correct diagnosis, since the indication of this type of treatment associated with an incorrect diagnosis leads to uncertain outcomes, creating false conclusions.

Quality of life of patients with facial prosthesis / Calidad de vida de los pacientes portadores de prótesis faciales

ABSTRACT. Objective: evaluate the quality of life and self-esteem of patients with facial deformities in the nasal, oculo-palpebral, and auricular areas using bucco-maxillofacial prostheses. Methods: descriptive study in users of facial prostheses who were rehabilitated at the dental clinic of the University Hospital of Brasilia in the period 2004-2011. The average age of patients was 63.8 years. The data collection instruments were the SF-36 questionnaire and a survey containing the Rosenberg Self-Esteem/EPM scale. Results: skin cancer was the predominant etiologic factor (n = 4). Most patients think that their deformity is visible to some degree (n = 4) and for three of them this fact impacts negatively the frequency to visit public places. The self-esteem scale yielded an average score of 4.6. In evaluating the quality of life by means of the SF-36 questionnaire, there were scores like pain (46.67) and emotional aspects (33.33). Conclusion: the results demonstrate the importance of conducting systematic followups to these patients. Most surveyed patients experienced an increase in quality of life, as well as a higher self-esteem, which can be explained by the functionality and aesthetics offered by the prosthesis. This study is part of an assessment program conducted with the intention of structuring multidisciplinary rehabilitation programs.

RESUMEN. Objetivo: evaluar la calidad de vida y la autoestima de los pacientes con deformidades faciales en la zona nasal, la zona óculopalpebral y la región auricular, portadores de prótesis bucomaxilofaciales. Métodos: estudio descriptivo en pacientes usuarios de prótesis faciales, los cuales fueron rehabilitados en la clínica odontológica del Hospital Universitario de Brasilia en el periodo 2004-2011. La edad media de los pacientes evaluados fue de 63,8 años. Como instrumentos de recolección de datos, en esta investigación se utilizaron el cuestionario SF-36 y una encuesta que contenía la Escala Visual de Autoestima de Rosenberg ̸EPM. Resultados: el cáncer de piel fue el factor etiológico predominante (n = 4). La mayoría de los pacientes consideró que su deformidad es visible en algún grado (n = 4), y para tres de ellos este hecho interfiere negativamente en la frecuencia de visitas a sitios públicos. En la escala de autoestima se obtuvo una puntuación media de 4,6. En la evaluación de la calidad de vida utilizando el cuestionario SF-36 se obtuvieron valores como dolor (46,67) y aspectos emocionales (33,33). Conclusión: los resultados demuestran la importancia de hacer un seguimiento sistemático a estos pacientes. En la mayoría de los pacientes encuestados se identificó un aumento en la calidad de vida, así como una mayor autoestima, lo cual puede explicarse por la funcionalidad y estética desempeñada por la prótesis. Esta es una propuesta de evaluación que se llevó a cabo con el fin de estructurar programas de rehabilitación multidisciplinar.

microRNA target prediction programs predict many false positives.

According to the current view, each microRNA regulates hundreds of genes. Computational tools aim at identifying microRNA targets, usually selecting evolutionarily conserved microRNA binding sites. While the false positive rates have been evaluated for some prediction programs, that information is rarely put forward in studies making use of their predictions. Here, we provide evidence that such predictions are often biologically irrelevant. Focusing on miR-223-guided repression, we observed that it is often smaller than inter-individual variability in gene expression among wild-type mice, suggesting that most predicted targets are functionally insensitive to that microRNA. Furthermore, we found that human haplo-insufficient genes tend to bear the most highly conserved microRNA binding sites. It thus appears that biological functionality of microRNA binding sites depends on the dose-sensitivity of their host gene and that, conversely, it is unlikely that every predicted microRNA target is dose-sensitive enough to be functionally regulated by microRNAs. We also observed that some mRNAs can efficiently titrate microRNAs, providing a reason for microRNA binding site conservation for inefficiently repressed targets. Finally, many conserved microRNA binding sites are conserved in a microRNA-independent fashion: Sequence elements may be conserved for other reasons, while being fortuitously complementary to microRNAs. Collectively, our data suggest that the role of microRNAs in normal and pathological conditions has been overestimated due to the frequent overlooking of false positive rates.

The gastrula transition reorganizes replication-origin selection in Caenorhabditis elegans.

Although some features underlying replication-origin activation in metazoan cells have been determined, little is known about their regulation during metazoan development. Using the nascent-strand purification method, here we identified replication origins throughout Caenorhabditis elegans embryonic development and found that the origin repertoire is thoroughly reorganized after gastrulation onset. During the pluripotent embryonic stages (pregastrula), potential cruciform structures and open chromatin are determining factors that establish replication origins. The observed enrichment of replication origins in transcription factor-binding sites and their presence in promoters of highly transcribed genes, particularly operons, suggest that transcriptional activity contributes to replication initiation before gastrulation. After the gastrula transition, when embryonic differentiation programs are set, new origins are selected at enhancers, close to CpG-island-like sequences, and at noncoding genes. Our findings suggest that origin selection coordinates replication initiation with transcriptional programs during metazoan development.

Argonaute proteins regulate HIV-1 multiply spliced RNA and viral production in a Dicer independent manner.

Argonaute (Ago) proteins associate with microRNAs (miRNAs) to form the core of the RNA-induced silencing complex (RISC) that mediates post-transcriptional gene silencing of target mRNAs. As key players in anti-viral defense, Ago proteins are thought to have the ability to interact with human immunodeficiency virus type 1 (HIV-1) RNA. However, the role of this interaction in regulating HIV-1 replication has been debated. Here, we used high throughput sequencing of RNA isolated by cross-linking immunoprecipitation (HITS-CLIP) to explore the interaction between Ago2 and HIV-1 RNA in infected cells. By only considering reads of 50 nucleotides length in our analysis, we identified more than 30 distinct binding sites for Ago2 along the viral RNA genome. Using reporter assays, we found four binding sites, located near splice donor sites, capable of repressing Luciferase gene expression in an Ago-dependent manner. Furthermore, inhibition of Ago1 and Ago2 levels in cells expressing HIV-1 led to an increase of viral multiply spliced transcripts and to a strong reduction in the extracellular CAp24 level. Depletion of Dicer did not affect these activities. Our results highlight a new role of Ago proteins in the control of multiply spliced HIV-1 transcript levels and viral production, independently of the miRNA pathway.

Retention and Mechanical Behavior of Attachment Systems for Implant-Retained Auricular Prostheses.

OBJECTIVE: Auricular prostheses are artificial substitutes for facial defects. The retention of these has often been a problem. This study aimed to evaluate the mechanical behavior of 3 retained auricular prosthetic connections when submitted to a mechanical cycling test. MATERIALS AND METHODS: Twelve samples with installed implants were obtained and divided into 3 groups according to their retention system with 4 samples in each group. I: bar-clip system; II: magnet system; and III: ball/o-ring system. Each of samples was submitted to the pull-out test during 3240 cycles (f = 0.5 Hz) to determine its tensile strength. The mechanical cycling test was performed using the servo-hydraulic machine MTS 810-Flex Test 40 (Eden Prairie, MN) that had a 2.5 mm shift at a 10 mm/s velocity. The retaining strength for each of the samples was obtained at 7 intervals. RESULTS: The tensile strength for the group retained by the bar-clip system (29.60 N) was higher with statistically significant difference (P < 0.05) when compared with the group retained by the ball/o-ring system (9.41 N) and magnets system (8.61 N) for all periods assessed. The ball/o-ring system showed loss of retention during the fatigue test (Kruskal-Wallis, chi-squared = 17.28; P < 0.01). CONCLUSIONS: The evaluated systems showed a tensile strength compatible with the clinical use and no fractures of the components were observed.

microRNAs and the evolution of complex multicellularity: identification of a large, diverse complement of microRNAs in the brown alga Ectocarpus.

There is currently convincing evidence that microRNAs have evolved independently in at least six different eukaryotic lineages: animals, land plants, chlorophyte green algae, demosponges, slime molds and brown algae. MicroRNAs from different lineages are not homologous but some structural features are strongly conserved across the eukaryotic tree allowing the application of stringent criteria to identify novel microRNA loci. A large set of 63 microRNA families was identified in the brown alga Ectocarpus based on mapping of RNA-seq data and nine microRNAs were confirmed by northern blotting. The Ectocarpus microRNAs are highly diverse at the sequence level with few multi-gene families, and do not tend to occur in clusters but exhibit some highly conserved structural features such as the presence of a uracil at the first residue. No homologues of Ectocarpus microRNAs were found in other stramenopile genomes indicating that they emerged late in stramenopile evolution and are perhaps specific to the brown algae. The large number of microRNA loci in Ectocarpus is consistent with the developmental complexity of many brown algal species and supports a proposed link between the emergence and expansion of microRNA regulatory systems and the evolution of complex multicellularity.

Bisphosphonate-related osteonecrosis of the jaw: a review of the literature.

Bisphosphonates (BPs) are a class of drugs used to treat osteoporosis and malignant bone metastasis. BPs show high binding capacity to the bone matrix, especially in sites of active bone metabolism. The American Society for Bone and Mineral Research defines BRONJ as "an area of exposed bone in the maxillofacial region that has not healed within 8 weeks after identification by a healthcare provider in a patient who is receiving or has been exposed to a bisphosphonate and has not had radiation therapy to the craniofacial region." Bisphosphonate-related osteonecrosis of the jaw (BRONJ) can adversely affect quality of life, as it may produce significant morbidity. The American Association of Oral and Maxillofacial Surgeons (AAOMS) considers as vitally important that information on BRONJ be disseminated to other dental and medical specialties. The purpose of this work is to offer a perspective on how dentists should manage patients on BPs, to show the benefits of accurately diagnosing BRONJ, and to present diagnostic aids and treatments strategies for the condition.

Targeting vertebrate intron-encoded box C/D 2'-O-methylation guide RNAs into the Cajal body.

Post-transcriptional pseudouridylation and 2'-O-methylation of splicesomal small nuclear ribonucleic acids (snRNAs) is mediated by box H/ACA and box C/D small Cajal body (CB)-specific ribonucleoproteins (scaRNPs), respectively. The WD-repeat protein 79 (WDR79) has been proposed to interact with both classes of modification scaRNPs and target them into the CB. The box H/ACA scaRNAs carry the common CAB box motif (consensus, ugAG) that is required for both WDR79 binding and CB-specific accumulation. Thus far, no cis-acting CB-localization element has been reported for vertebrate box C/D scaRNAs. In this study, systematic mutational analysis of the human U90 and another newly identified box C/D scaRNA, mgU2-47, demonstrated that the CB-specific accumulation of vertebrate intron-encoded box C/D scaRNAs relies on GU- or UG-dominated dinucleotide repeat sequences which are predicted to form the terminal stem-loop of the RNA apical hairpin. While the loop nucleotides are unimportant, the adjacent terminal helix that is composed mostly of consecutive G.U and U.G wobble base-pairs is essential for CB-specific localization of box C/D scaRNAs. Co-immunoprecipitation experiments confirmed that the newly identified CB localization element, called the G.U/U.G wobble stem, is crucial for in vivo association of box C/D scaRNPs with WDR79.