GATA2 mitotic bookmarking is required for definitive haematopoiesis.

In mitosis, most transcription factors detach from chromatin, but some are retained and bookmark genomic sites. Mitotic bookmarking has been implicated in lineage inheritance, pluripotency and reprogramming. However, the biological significance of this mechanism in vivo remains unclear. Here, we address mitotic retention of the hemogenic factors GATA2, GFI1B and FOS during haematopoietic specification. We show that GATA2 remains bound to chromatin throughout mitosis, in contrast to GFI1B and FOS, via C-terminal zinc finger-mediated DNA binding. GATA2 bookmarks a subset of its interphase targets that are co-enriched for RUNX1 and other regulators of definitive haematopoiesis. Remarkably, homozygous mice harbouring the cyclin B1 mitosis degradation domain upstream Gata2 partially phenocopy knockout mice. Degradation of GATA2 at mitotic exit abolishes definitive haematopoiesis at aorta-gonad-mesonephros, placenta and foetal liver, but does not impair yolk sac haematopoiesis. Our findings implicate GATA2-mediated mitotic bookmarking as critical for definitive haematopoiesis and highlight a dependency on bookmarkers for lineage commitment.

First record of a sea anemone of genus Bunodosoma with two oral discs: a clue to asexual reproduction?

Different reproductive strategies (both sexual and asexual) that may occur in individuals of the same species may explain the success of sea anemones (Cnidaria:Actiniaria) in colonizing different ecosystems around the planet. Here we provide the first record of a Bunodosoma specimen with two oral discs and discuss the possibility of asexual reproduction in this genus. One individual of Bunodosoma cangicum was found in Carneiros beach (Pernambuco state, Brazil) with two fully individualized oral discs but with column and pedal disc still connected. Our finding may represent a clue to longitudinal fission in Bunodosoma. Yet, despite the number of different studies carried out so far, the absence of data regarding asexual reproduction in Bunodosoma suggests that it might be rare in this genus, occurring only in sporadic cases such as recovery from injury.

Familial lecithin-cholesterol acyltransferase deficiency: If so rare, why so frequent in the state of Piauí, northeastern Brazil?

Lecithin-cholesterol acyltransferase (LCAT), an enzyme that participates in lipoprotein metabolism, plays an important role in cholesterol homeostasis. Mutations in the LCAT gene can cause two rare genetic disorders: familial LCAT deficiency (FLD), which is characterized by corneal opacities, normocytic anemia, dyslipidemia, and proteinuria progressing to chronic renal failure, and fish-eye disease (FED), which causes dyslipidemia and progressive corneal opacities. Herein, we report six suspected cases of FLD in the backlands of Piauí, located in northeast Brazil. A genetic diagnosis was performed in index cases. Among these, a further investigation was performed to identify new cases in the families. In addition, molecular analyses were performed to verify the levels of consanguinity within families and the existence of a genetic relationship between them. All six index cases were confirmed as FLD with an identical mutation (c.803G > A, p.R268H). The genetic investigation confirmed another 7 new cases of FLD, 52 heterozygous and 6 individuals without mutations. The rate of consanguinity revealed that marriages within the family did not contribute to the high number of FLD cases within the restricted region. The elders of each family (patriarchs and matriarchs) were subjected to a kinship analysis and were more genetically related to each other than the control group. Bayesian analysis was implemented to confirm the hypothesis of connectivity among patriarchs and matriarchs and indicated that they were genetically more related to each other than would be randomly expected, thus suggesting the occurrence of a possible founder effect in these families.

MRCKα Is Dispensable for Breast Cancer Development in the MMTV-PyMT Model.

MRCKα is a ubiquitously expressed serine/threonine kinase involved in cell contraction and F-actin turnover, which is highly amplified in human breast cancer and part of a gene expression signature for bad prognosis. Nothing is known about the in vivo function of MRCKα. To explore MRCKα function in development and in breast cancer, we generated mice lacking a functional MRCKα gene. Mice were born close to the Mendelian ratio and showed no obvious phenotype including a normal mammary gland formation. Assessing breast cancer development using the transgenic MMTV-PyMT mouse model, loss of MRCKα did not affect tumor onset, tumor growth and metastasis formation. Deleting MRCKα and its related family member MRCKß in two triple-negative breast cancer cell lines resulted in reduced invasion of MDA-MB-231 cells, but did not affect migration of 4T1 cells. Further genomic analysis of human breast cancers revealed that MRCKα is frequently co-amplified with the oncogenes ARID4B and AKT3 which might contribute to the prognostic value of MRCKα expression. Collectively, these data suggest that MRCKα might be a prognostic marker for breast cancer, but probably of limited functional importance.

ACK1 is dispensable for development, skin tumor formation, and breast cancer cell proliferation.

Activated Cdc42-associated kinase 1 (ACK1), a widely expressed nonreceptor tyrosine kinase, is often amplified in cancer and has been shown to interact with Cell division cycle 42 (Cdc42), Epidermal growth factor receptor (EGFR), and several other cancer-relevant molecules, suggesting a possible role for ACK1 in development and tumor formation. To directly address this scenario, we generated mice lacking a functional ACK1 gene (ACK1 ko) using CRISPR genome editing. ACK1 ko mice developed normally, displayed no obvious defect in tissue maintenance, and were fertile. Primary ACK1-null keratinocytes showed normal phosphorylation of EGFR, but a tendency toward reduced activation of AKT serine/threonine kinase 1 (Akt) and Mitogen-activated protein kinase 1 (Erk). DMBA/TPA-induced skin tumor formation did not reveal significant differences between ACK1 ko and control mice. Deletion of the ACK1 gene in the breast cancer cell lines MDA-MB-231, 67NR, MCF7, 4T1, and T47D caused no differences in growth. Furthermore, EGF-induced phosphorylation kinetics of Erk, Akt, and p130Cas were not detectably altered in T47D cells by the loss of ACK1. Finally, loss of ACK1 in MDA-MB-231 and T47D breast cancer cells had a very limited or no effect on directed cell migration. These data do not support a major role for ACK1 in Cdc42 and EGFR signaling, development, or tumor formation.

Magnetic resonance imaging features of COVID-19-related cranial nerve lesions.

The complete features of the neurological complications of coronavirus disease 2019 (COVID-19) still need to be elucidated, including associated cranial nerve involvement. In the present study we describe cranial nerve lesions seen in magnetic resonance imaging (MRI) of six cases of confirmed COVID-19, involving the olfactory bulb, optic nerve, abducens nerve, and facial nerve. Cranial nerve involvement was associated with COVID-19, but whether by direct viral invasion or autoimmunity needs to be clarified. The development of neurological symptoms after initial respiratory symptoms and the absence of the virus in the cerebrospinal fluid (CSF) suggest the possibility of autoimmunity.

pHLA3D: Updating the database of predicted three-dimensional structures of HLA with HLA-DR, HLA-DQ and HLA-DP molecules.

To improve the availability of three-dimensional (3D) structures of HLA molecules, we created the pHLA3D database. In its first version, we modeled and published 106 3D structures of HLA class I molecules from the HLA-A, HLA-B, and HLA-C loci. This paper presents an update of this database, providing more 127 3D structures of HLA class II molecules (41 DR, 42 DQ, and 44 DP), predicted via homology modeling with MODELLER and SWISS-MODEL. These new 3D structures of HLA class II molecules are now freely available at pHLA3D (www.phla3d.com.br) for immunologists and other researchers working with HLA molecules.

pHLA3D: An online database of predicted three-dimensional structures of HLA molecules.

HLA epitope analysis emerged as a strategy to determine alloimmune risk in solid organ transplantation. However, it requires not only knowledge on HLA amino acids sequences, but also on HLA three-dimensional structures. Unfortunately, the number of structures available is still unsatisfactory. This work reports the modelling of 106 heterotrimeric (alpha chain + ß2M + peptide) HLA class I molecules. The models were generated by homology modelling using Modeller, refined using GalaxyRefine server, heterodimerized with Swiss-PDB Viewer and, finally, assessed as to their structural quality through Dali server. The final structures were made available through a free online database, pHLA3D (www.phla3d.com.br), developed in Ruby language using the Ruby on Rails web framework. Structural parameters were similar between refined molecules and their templates. The new database may improve HLA epitope analysis and better guide risk assessment in solid organ transplantation setting.

Remote post-conditioning and allopurinol reduce ischemia-reperfusion injury in an infra-renal ischemia model.

BACKGROUND:: The aim of this study was to evaluate the effects of the antioxidant allopurinol and ischemic post-conditioning on the deleterious effects of ischemia followed by reperfusion (I/R) in a standardized model of ischemia involving infra-renal aortic occlusion in rats. METHODS:: The animals were randomly divided into five groups: (A) animals not subjected to ischemia; (B) animals subjected to 2 h of ischemia and reperfusion only once; (C) animals given an allopurinol dose by gavage, then subjected to 2 h of ischemia and reperfusion only once; (D) animals subjected to 2 h of ischemia and post-conditioning and (E) animals that received allopurinol, then subjected to 2 h of ischemia and post-conditioning. The blood samples and small intestine segments were harvested for analysis after 3 days. RESULTS:: The protective effects of the use of allopurinol and ischemic post-conditioning were observed by measuring aspartate aminotransferase, alanine aminotransferase and lactate levels. The benefits of post-conditioning were evident from the total antioxidant capacity and creatinine levels, but these could not ascertain any positive effects of allopurinol. The histological analysis of mesentery revealed that both methods were effective in minimizing the harmful effects of the ischemia and reperfusion process. CONCLUSION:: Individual protocols significantly reduced I/R systemic injuries, but no additional protection was observed when the two strategies were combined.

Novel recombinant multiepitope proteins for the detection of anti-Cryptococcus antibodies.

AIM: To produce and test recombinant multiepitope proteins as an alternative assay for the serological diagnosis of cryptococcosis. MATERIALS & METHODS: Previously, synthetic peptides were used to detect anti-Cryptococcus antibodies, and in silico analyses showed that the union of peptides would improve the results. Here, the coding sequences of these peptides were assembled into synthetic genes. Four genes have been cloned and expressed in Escherichia coli, producing recombinant multiepitope proteins: proteins A, B, C and D. RESULTS: All constructs yielded good results; however, protein D showed the best results, with a sensitivity of 88.57% and specificity of 100%. CONCLUSION: The multiepitope proteins were shown to be potential antigens for the diagnosis of cryptococcosis in an attempt to detect anti-Cryptococcus antibodies.

Titanium versus polyetheretherketone implants for vertebral body replacement in the treatment of 77 thoracolumbar spinal fractures.

BACKGROUND: Titanium and polyetheretherketone (PEEK) implants have been used in spinal surgery with low rejection rates. Compared to titanium, PEEK has many advantages, including a density more similar to that of bone, radiolucency, and a lack of artifacts in computed tomography (CT) and magnetic resonance imaging (MRI). In this study, we evaluated the effectiveness of PEEK cages as an alternative to titanium for bone fusion after fractures of the thoracolumbar spine. We also propose a classification to the impaction index. METHODS: We evaluated 77 patients with fractures of the thoracic or lumbar spine who were treated by anterior fixation with titanium cages (TeCorp®) in 46 (59.7%) patients or PEEK (Verte-stak®) in 31 (40.3%) patients from 2006 to 2012 (Neurological Hospital of Lyon). RESULTS: The titanium group achieved 100% fusion, and the PEEK group achieved 96.3% fusion. The titanium systems correlated with higher impact stress directed toward the lower and upper plateaus of the fused vertebrae; there were no nonunions for those treated with titanium group. Nevertheless, there was only one in the PEEK group. There was no significant difference in the pain scale outcomes for patients with ±10 degrees of the sagittal angle. Statistically, it is not possible to associate the variation of sagittal alignment or the impaction with symptoms of pain. The complication rate related to the implantation of cages was low. CONCLUSIONS: Titanium and PEEK are thus equally effective options for the reconstruction of the anterior column. PEEK is advantageous because its radiolucency facilitates the visualization of bone bridges.

Abscesso subperiosteal com extensão epidural devido à rinossinusite aguda em criança de dez anos / Subperiosteal abscess with epidural extension due to acute rhinosinusitis in a ten-year-old child

RESUMO A rinossinusite aguda é uma das afecções mais prevalentes das vias aéreas superiores. Fatores anatômicos presentes em crianças e jovens propiciam o aparecimento de complicações orbitárias. Embora mais raras, as complicações intracranianas das rinossinusites perfazem um grau alto de letalidade, são mais comuns em pacientes acima de sete anos, e devem ser tratadas por uma equipe multidisciplinar.

ABSTRACT Acute rhinosinusitis is one of the most prevalent diseases of the upper airways. Anatomical factors present in children and young people allow for the onset of orbital complications. Although rare, intracranial complications of sinusitis account for a high degree of lethality, are more common in patients over the age of seven, and should be treated by a multidisciplinary team.

Linfoma primário de efusão em mulher com síndrome da imunodeficiênciaadquirida (SIDA), derrame pleural e ascite: diagnóstico citológico e imunocitoquímico / Primary effusion lymphoma in women with acquired immunodeficiency syndrome (AIDS), pleural effusion, and ascites: cytological diagnosis and immunocytochemistry

O linfoma primário de efusão é tipo raro de forma não Hodgkin que se manifesta com derrame cavitário, em geral associado à imunodepressão. Associa-se à infecção pelo vírus herpes humano 8 (HHV-8), fato necessário para que seja confirmado seu diagnóstico.Este relato descreve o linfoma primário de efusão, diagnosticado por meio de citologia e imunocitoquímica, em paciente com manifestação não usual de comprometimento de duas cavidades serosas. Seu prognóstico é reservado e seu tratamento é limitado.

The primary effusion lymphoma is a rare type of non-Hodgkin form that manifests with cavity stroke, usually associated with immunosuppression. It is associated with infection with the human herpes virus 8 (HHV-8), a necessary fact to confirm the diagnosis. This report describes the primary effusion lymphoma, diagnosed by cytology and immunocytochemistry,in patients with the unusual manifestation of involvement of two serouscavities. Its prognosis is poor, and treatment is limited.

Immunoreactivity of synthetic peptides derived from proteins of Cryptococcus gattii.

AIM: To determine the immunoreactivity of synthetic Cryptococcus-derived peptides. MATERIALS & METHODS: A total of 63 B-cell epitopes from previously identified Cryptococcus gattii immunoreactive proteins were synthesized and evaluated as antigens in ELISAs. The peptides were first evaluated for their ability to react against sera from immunocompetent subjects carrying cryptococcal meningitis. Peptides that yielded high sensitivity and specificity in the first test were then retested with sera from individuals with other fungal pathologies for cross-reactivity determination. RESULTS: Six of 63 synthetic peptides were recognized by antibodies in immunoassays, with a specificity of 100%, sensitivity of 78% and low cross-reactivity. CONCLUSION: We successfully determined the immunoreactivity of selected synthetic peptides of C. gattii derived proteins.

Transposition of cephalic vein to rescue hemodialysis access arteriovenous fistula and treat symptomatic central venous obstruction / Transposição de veia cefálica para salvamento de fístula arteriovenosa de hemodiálise e tratamento de obstrução venosa central sintomática

It is known that stenosis or central venous obstruction affects 20 to 50% of patients who undergo placement of catheters in central veins. For patients who are given hemodialysis via upper limbs, this problem causes debilitating symptoms and increases the risk of loss of hemodialysis access. We report an atypical case of treatment of a dialysis patient with multiple comorbidities, severe swelling and pain in the right upper limb (RUL), few alternative sites for hemodialysis vascular access, a functioning brachiobasilic fistula in the RUL and severe venous hypertension in the same limb, secondary to central vein occlusion of the internal jugular vein and right brachiocephalic trunk. The alternative surgical treatment chosen was to transpose the RUL cephalic vein, forming a venous necklace at the anterior cervical region, bypassing the site of venous occlusion. In order to achieve this, we dissected the cephalic vein in the right arm to its junction with the axillary vein, devalved the cephalic vein and anastomosed it to the contralateral external jugular vein, providing venous drainage to the RUL, alleviating symptoms of venous hypertension and preserving function of the brachiobasilic fistula.

Sabemos que estenose ou obstrução venosa central ocorre em 20 a 50% dos pacientes que são submetidos à colocação de cateter em veias centrais. Nos pacientes que realizam hemodiálise pelos membros superiores, este problema causa sintomas debilitantes e um grande risco de perda do acesso para hemodiálise. Relatamos um caso atípico de tratamento em um paciente dialítico com múltiplas comorbidades, queixa de dor e edema severo do membro superior direito (MSD), escassas alternativas de acessos vasculares para hemodiálise e fístula braquiobasílica funcionante do MSD associada à severa hipertensão venosa deste membro, secundária à oclusão venosa central da veia jugular interna e do tronco braquiocefálico direito. O tratamento cirúrgico alternativo foi a transposição da veia cefálica do MSD, formando colar venoso na região cervical anterior, resultando em um bypass sobre o sítio venoso ocluído. Para isso, realizamos a dissecção da veia cefálica no braço direito até a sua junção com a veia axilar, devalvulamos e anastomosamos a veia cefálica na veia jugular externa contralateral, permitindo a drenagem venosa do MSD, aliviando os sintomas da hipertensão venosa e mantendo a fístula braquiobasílica funcionante.

Immunoproteomics and immunoinformatics analysis of Cryptococcus gattii: novel candidate antigens for diagnosis.

AIM: To identify immunoreactive proteins of Cryptococcus gattii genotype VGII and their B-cell epitopes. MATERIALS & METHODS: We combined 2D gel electrophoresis, immunoblotting and mass spectrometry to identify immunoreactive proteins from four strains of C. gattii genotype VGII (CG01, CG02, CG03 and R265). Next, we screened the identified proteins to map B-cell epitopes. RESULTS: Sixty-eight immunoreactive proteins were identified. The strains and the number of proteins we found were: CG01 (12), CG02 (12), CG03 (18) and R265 (26). In addition, we mapped 374 peptides potentially targeted by B cells. CONCLUSION: Both immunoreactive proteins and B-cell epitopes of C. gattii genotype VGII that were potentially targeted by a host humoral response were identified. Considering the evolutionary relevance of the identified proteins, we may speculate that they could be used as the initial targets for recombinant protein and peptide synthesis aimed at the development of immunodiagnostic tools for cryptococcosis.

Efficacy and safety of a porcine collagen sponge for cranial neurosurgery: a prospective case-control study.

OBJECTIVE: The use of dural grafts is very useful when primary dural closure cannot be achieved. Our primary objective was to study the incidence of postoperative cerebrospinal fluid leak, including fistula and pseudomeningocele, and postoperative infection by comparing autologous material and a new collagen graft. MATERIALS AND METHODS: A prospective nonrandomized study with a new collagen-based product derived from porcine cells (Peridry) was performed. It was used for dural replacement in 50 patients who underwent a variety of neurosurgical procedures requiring the use of a dural graft. These results were compared with a control group of 50 patients who were treated with autologous duraplasty material. The follow-up period was 3 months. RESULTS: Postoperative overall cerebrospinal fluid fistula occurred in 6% of both groups. No patient in the collagen group developed any sort of infection. One patient in the control developed osteomyelitis in the bone flap. CONCLUSION: The new collagen-based product derived from porcine cells (Peridry), compared with an autologous tissue, is safe, effective, easy to use, as well as time saving in cranial neurosurgery.

Ventriculite por acinetobacter baumannii multiresistente / Multiresistant acinetobacter baumannii ventriculitis

Entre as bactérias Gram-negativas, o Acinetobacter sp tornou-se um importante patógeno hospitalar, devido ao aumento do número de cepas multi-resistentes. Esta espécie é responsável por um número crescente de infecções pósoperatórias caracterizadas por alta mortalidade. A ocorrência de bactérias Gram-negativas multirresistentes levou a um aumento no número de infecções do sistema nervoso central. Em particular, a ocorrência de bactérias resistentes à cefalosporinas de quarta geração e carbapenênicos resultou em uma redução significativa de opções terapêuticas para o tratamento destas infecções. Acinetobacter baumannii é um importante agente hospitalar e sua resistência aos antibióticos mais modernos aumenta a cada dia, o que é uma grave ameaça aos pacientes infectados. Descrevemos um caso de um paciente submetido a neurocirurgia e colocação de derivação ventricular externa evoluindo com ventriculite por Acinetobacter baumannii resistente a cefalosporinas de quarta geração e meropenem.

Pseudotumoral form of cerebral Schistosomiasis mansoni.

OBJECTIVE: To describe published cases of cerebral mansoni schistosomiasis and three others and discuss the diagnosis and treatment of cerebral pseudotumoral schistosomiasis. CASE DESCRIPTIONS: In case 1, a 20-year-old man presented with occipital headache, intense dizziness, visual alterations, nausea, decreased appetite, and asthenia. Cranial computed tomography (CT) revealed an expansive cerebellar lesion in the right hemisphere with no contrast enhancement. The patient had complete resection of the lesion. Anatomicopathological examination revealed a schistosomal granuloma. In case 2, a 22-year-old man presented with generalized tonic-clonic seizure. Cranial CT and magnetic resonance imaging (MRI) revealed an expansive bilateral middle frontal lesion, with contrast uptake close to the cingulate gyrus and corpus callosum. The patient underwent left frontal craniotomy, and an interhemispheric approach was used to resect part of the lesion. In case 3, a 32-year-old man presented with generalized tonic-clonic seizures. Cranial CT showed a hyperdense intense intracranial expansive lesion that presented contrast uptake in the left temporal region. The patient had complete resection of the lesion. CONCLUSIONS: A surgical approach with lesion resection or stereotaxic biopsy is warranted to determine the diagnosis definitively. Antiparasitic drugs must be administered to complete treatment.

Papillary tumor of the pineal region.

Tumors of the pineal region are rare in adulthood, accounting for approximately 1% of intracranial neoplasms in this age range. Papillary tumor of the pineal region (PTPR) was first described by Jouvet et al. in 2003. In 2007, PTPR was included by the World Health Organization as a distinct entity in their new classification of central nervous system tumors. We report a case of PTPR, and undertake a literature review of this rare neuroepithelial neoplasm. Patients with PTPR need long-term follow up, and new cases should be well documented so that we can gain a better understanding of this neoplasm.