Analysis of errors in histology by root cause analysis: a pilot study

Introduction. The study objective is to evaluate critical points in the process of pre-analytical histology in an Anatomic Pathology laboratory. Errors are an integral part of human systems, includ- ing the complex system of Anatomic Pathology. Previous studies focused on errors committed in diagnosis and did not consider the issues related to the histology preparation of routine processes. Methods. Root Cause Analysis was applied to the process of histology preparation in order to identify the root cause of each previously identified problem. The analysis started by defining an ?a priori? list of errors that could occur in the histology prepara- tion processes. During a three-month period, a trained technician tracked the errors encountered during the process and reported them on a form. ?Fishbone? diagram and ?Five whys? methods were then applied. Results. 8,346 histological cases were reviewed, for which 19,774 samples were made and from which 29,956 histologies were pre- pared. 132 errors were identified. Errors were detected in each phase: accessioning (6.5%), gross dissecting (28%), processing (1.5%), embedding (4.5%), tissue cutting and slide mounting (23%), coloring, (1.5%), labeling and releasing (35%). Discussion. Root cause analysis is effective and easy to use in clinical risk management. It is an important step for the identifi- cation and prevention of errors, that are frequently due to multi- ple causes. Developing operators? awareness of their central role in the risk management process is possible by targeted training. Furthermore, by highlighting the most relevant points of interest, it is possible to improve both the methodology and the procedural safety

Efeito de operações de preparo de solo em pós-colheita sobre Sternechus subsignatus (Col.: Curculionidae).

Sternechus subsignatus (Baheman), conhecido por tamanduá-da-soja, é uma praga muito prejudicial à cultura da soja no cerrado do Oeste da Bahia. Nos anos de 2005 (18/maio a 16/novembro) e 2006 (15/junho a 08/novembro), em Formosa do Rio Preto-BA, avaliou-se o efeito dos seguintes métodos de preparo do solo em póscolheita no controle deste inseto: T1) sem preparo de solo; T2) subsolador; T3) subsolador + grade intermediária (30 dias após a primeira operação); T4) arado de aiveca; T5) arado de aiveca + grade intermediária (30 dias após a primeira operação), e T6) grade intermediária. A infestação média da área era de 35,1 e 25,4 larvas/m2 a 0-20cm de profundidade do solo no momento da instalação dos experimentos em 2005 e 2006, respectivamente, com 80% das larva na faixa de 0-10cm. O delineamento foi o de blocos ao acaso, com quatro repetições, em parcelas de 144,0m2 (12,0m x 12,0m). O número de larvas/m2 foi determinado em três pontos/parcela (1,0m x 0,5m) nas profundidades de 0-5cm, 5-10cm, 10-15cm e 15-20cm. Os dados foram analisados pelo teste de Scott-Knott (1974), a 5% de probabilidade, e controle por Abbott (1925). O efeito dos tratamentos sobre a população de larvas foi consistente, com grande similaridade de resultados nos dois anos para todos os tratamentos. Os valores médios de controle para os tratamentos nos dois anos, e seus respectivos valores anuais foram: T2 (38% = 39,6% e 36,2%), T3 (66% = 61,3% e 70,8%), T4 (58% = 54,1% e 61,7%), T5 (81% = 76,3% e 85,9%) e T6 (20% = 24,7% e 14,7%). O impacto dos tratamentos T3, T4 e T5 sobre a população do inseto foi mais evidentes 80 dias após o início do ensaio. A segunda operação com grade incrementou significativamente o controle dos insetos iniciado com o subsolador e aiveca. Operação de preparo de solo em pós-colheita, com implementos corretos, é uma tática de controle viável no manejo dos focos populações de S. subsignatus no solo

Inability to switch from ARID1A-BAF to ARID1B-BAF impairs exit from pluripotency and commitment towards neural crest formation in ARID1B-related neurodevelopmental disorders

Mutations in the ARID1B subunit of the BAF chromatin remodeling complex are associated with the neurodevelopmental Coffin-Siris syndrome. Here the authors reveal that there is a transition from ARID1A-containing complexes to ARID1B during cranial neural crest cell differentiation that is impaired in Coffin-Siris patient-derived cells, which is important for exit from pluripotency.Subunit switches in the BAF chromatin remodeler are essential during development. ARID1B and its paralog ARID1A encode for mutually exclusive BAF subunits. De novo ARID1B haploinsufficient mutations cause neurodevelopmental disorders, including Coffin-Siris syndrome, which is characterized by neurological and craniofacial features. Here, we leveraged ARID1B(+/-) Coffin-Siris patient-derived iPSCs and modeled cranial neural crest cell (CNCC) formation. We discovered that ARID1B is active only during the first stage of this process, coinciding with neuroectoderm specification, where it is part of a lineage-specific BAF configuration (ARID1B-BAF). ARID1B-BAF regulates exit from pluripotency and lineage commitment by attenuating thousands of enhancers and genes of the NANOG and SOX2 networks. In iPSCs, these enhancers are maintained active by ARID1A-containing BAF. At the onset of differentiation, cells transition from ARID1A- to ARID1B-BAF, eliciting attenuation of the NANOG/SOX2 networks and triggering pluripotency exit. Coffin-Siris patient cells fail to perform the ARID1A/ARID1B switch, and maintain ARID1A-BAF at the pluripotency enhancers throughout all stages of CNCC formation. This leads to persistent NANOG/SOX2 activity which impairs CNCC formation. Despite showing the typical neural crest signature (TFAP2A/SOX9-positive), ARID1B-haploinsufficient CNCCs are also aberrantly NANOG-positive. These findings suggest a connection between ARID1B mutations, neuroectoderm specification and a pathogenic mechanism for Coffin-Siris syndrome.Therapeutic cell differentiatio

Árvores nativas para sistemas agrícolas sustentáveis e para restauração ecológica na região Noroeste do Rio Grande do Sul.

Este Documento reúne um conjunto de 45 espécies de árvores e arvoretas nativas da Floresta Estacional, as quais são recomendadas para ecossistemas naturais e cultivados no noroeste do Rio Grande do Sul, com ênfase às regiões situadas na bacia hidrográfica do rio Uruguai. Ao mesmo tempo, essas espécies são passíveis de utilização em outras regiões do estado, a depender da adaptação ambiental e/ou genética nesses locais. A publicação é resultado do Projeto Rede RestaurAção, executado entre janeiro de 2018 e janeiro de 2021, por meio do convênio entre Embrapa Clima Temperado, Empresa Nacional de Energia Elétrica (Enel) e a Fundação de Apoio à Pesquisa e Desenvolvimento Agropecuário Edmundo Gastal (Fapeg), o qual reuniu um conjunto de outros parceiros essenciais à realização de suas ações. Foram eles: a Associação Regional de Educação, Desenvolvimento e Pesquisa (Arede), a Cooperativa Agropecuária dos Agricultores Familiares de Tenente Portela (CooperFamiliar), a Universidade Regional do Noroeste do Estado do Rio Grande do Sul (Unijuí), a Sociedade Educacional Três de Maio (Setrem), a Emater/RS e a Secretaria do Ambiente e Desenvolvimento Sustentável do RS (Sema/RS). Ainda, em relação aos Objetivos de Desenvolvimento Sustentável (ODS), definidos pela Organização das Nações Unidas (ONU) para todos os países, esta publicação contempla o ODS 15: ?Vida terrestre: proteger, recuperar e promover o uso sustentável dos ecossistemas terrestres, gerir de forma sustentável as florestas, combater a desertificação, deter e reverter a degradação da Terra e deter a perda da biodiversidade?.ODS 15

ROLE OF THE NOTCH LIGAND JAGGED1 IN ZEBRAFISH THYROID DEVELOPMENT

Notch signaling is the result of the complex interaction of various Delta and Jagged ligands and their Notch receptors. This signal is known to have a relevant and expanding role in cell fate determination, survival and proliferation during embryonic development. Though Notch pathway is known to have a fundamental role in different endocrine systems (i.e. anterior pituitary and pancreatic cell specification), its possible involvement in thyroid development has never been studied. Our previous gene expression analysis in a thyroid cell line revealed the presence of several transcripts belonging to the Jagged1-Notch pathway, thus suggesting its potential role in thyroid development. Then, we designed a series of experiments aimed at confirming these preliminary data in vitro and understand the role of Notch signaling in thyroid development. For this purpose, we took advantage of zebrafish, a versatile experimental model that has been shown to largely recapitulate the early events of mammalian thyroid gland organogenesis. Gene expression analysis and experimental manipulations of zebrafish embryos were performed to identify the possible contribution of Jagged1 ligands to the thyroid development. Loss of Notch receptors activation in mind bomb (mib) mutants or DAPT-treated embryos led to an increased number of thyroid primordium cells. Conversely, Notch gain of function resulted in an impaired expression of the early thyroid marker genes (nkx2.1a; tg) in vivo. Given that the entire pharyngeal architecture was seriously affected by these genetic conditions, the observed thyroid phenotypes could come from either cell-autonomous or non cell-autonomous activities of Notch signaling. In order to understand if Notch signaling plays a direct role in thyroid development, we then analyzed the co-expression pattern of the zebrafish jag1a and jag1b, orthologues of the mammalian Jagged1 ligand, and thyroid markers by double whole-mount in situ hybridization. These experiments show the co-expression of these ligands in the thyroid region, at different times. Next, to understand the biological role of these ligands, we took advantage of both morpholino knock-down strategy and specific zebrafish mutants. Thyroid development was impaired in several of these conditions, as shown by the altered expression of thyroid differentiation markers tg and slc5a5 and T4 production at different time points. As a consequence, a defective function of Jagged1 ligand might constitute a novel mechanism involved in the pathogenesis of congenital hypothyroidism

Targeting the cdk6 dependence of ph+ acute lymphoblastic leukemia

Ph+ ALL is a poor-prognosis leukemia subtype driven by the BCR-ABL1 oncogene, either the p190-or the p210-BCR/ABL isoform in a 70:30 ratio. Tyrosine Kinase inhibitors (TKIs) are the drugs of choice in the therapy of Ph+ ALL. In combination with standard chemotherapy, TKIs have markedly improved the outcome of Ph+ ALL, in particular if this treatment is followed by bone marrow transplantation. However, resistance to TKIs develops with high frequency, causing leukemia relapse that results in <5-year overall survival. Thus, new therapies are needed to address relapsed/TKI-resistant Ph+ ALL. We have shown that expression of cell cycle regulatory kinase CDK6, but not of the highly related CDK4 kinase, is required for the proliferation and survival of Ph+ ALL cells. Comparison of leukemia suppression induced by treatment with the clinically-approved dual CDK4/6 inhibitor palbociclib versus CDK6 silencing revealed that the latter treatment was markedly more effective, probably reflecting inhibition of CDK6 kinase-independent effects. Thus, we developed CDK4/6-targeted proteolysis-targeting chimeras (PROTACs) that preferentially degrade CDK6 over CDK4. One compound termed PROTAC YX-2-107, which degrades CDK6 by recruiting the Cereblon ubiquitin ligase, markedly suppressed leukemia burden in mice injected with de novo or TKI-resistant Ph+ ALL. The effect of PROTAC YX-2-107 was comparable or superior to that of palbociclib. The development of CDK6-selective PROTACs represents an effective strategy to exploit the “CDK6 dependence” of Ph+ ALL cells while sparing a high proportion of normal hematopoietic progenitors that depend on both CDK6 and CDK6 for their survival. In combination with other agents, CDK6-selective PROTACs may be valuable components of chemotherapy-free protocols for the therapy of Ph+ ALL and other CDK6-dependent hematological malignancies

Thyroid gland development and function in the zebrafish model

Thyroid development has been intensively studied in the mouse, where it closely recapitulates the human situation. Despite the lack of a compact thyroid gland, the zebrafish thyroid tissue originates from the pharyngeal endoderm and the main genes involved in its patterning and early development are conserved between zebrafish and mammals. In recent years, the zebrafish has become a powerful model not only for the developmental biology studies, but also for large-scale genetic analyses and drug screenings, mostly thanks to the ease with which its embryos can be manipulated and to its translucent body, which allows in vivo imaging. In this review we will provide an overview of the current knowledge of thyroid gland origin and differentiation in the zebrafish. Moreover, we will consider the action of thyroid hormones and some aspects related to endocrine disruptors

PKA II mediates cAMP-dependent phosphorylation of CREB and activation of gene transcription in differentiated rat thyroid cells

Thyroid stimulating hormone (TSH) is the key modulator of thyroid cell function. Binding of TSH to its receptor (TSHR) results in an increase of cAMP intracellular concentration, which activates protein kinase A (PKA) to phosphorylate CREB and other substrates. Eventually, this leads to gene transcription modification associated with both proliferation and differentiation. Two major types of PKA (PKA I and II) exist in mammal cells, but their specific role in TSH signaling is poorly understudy so far. Therefore, the aim of this study was to dissect PKA I/II effects on gene transcription in thyroid cells. In order to selectively activate PKA I or II, pairs of cAMP analogs with different affinities for sites A and B of PKA regulatory subunits were used. The effect of PKA I or II activation on the transcription of early genes, genes involved in cell replication and genes associated with thyroid differentiated function was evaluated in FRTL5 cells by quantitative real-time PCR. The results indicate that PKA II, but not PKA I, stimulation can mimic the effect of TSH treatment on these genes. In addition, pre-treatment with a PKA II inhibitor was able to abolish the effect of TSH. Since CREB is the major transcriptional mediator of cAMP signal, we also evaluated the effect of PKA I or II activation on CREB phosphorylation. Also in this case, only PKA II activation lead to efficient CREB phosphorylation, and TSH-dependent activation was abolished by pre-incubation with the PKA II inhibitor. Finally, silencing of PKA regulatory subunit 2B (Prkar2b) by RNA interference resulted in down-regulation of proliferative genes. These results indicate that PKA II is the principal mediator of cAMP transcriptional effects in thyroid cells. Further studies are needed to evaluate the role of PKA I, which could play a role in post-transcriptional modifications rather than directly affecting gene transcription