Bruno Carvalho

New primers for the amplification and sequencing of nuclear loci in a taxonomically wide set of reptiles and amphibians

We report new primers for the amplification and sequencing of 11 nuclear markers in squamate reptiles and anuran amphibians (five in squamates, six in anurans). Ten out of the 11 loci are introns (three of which are linked) that were amplified using an exon-primed, intron-crossing (EPIC) PCR strategy, whereas an eleventh locus spans part of a protein-coding gene. Squamate and anuran primers were initially developed for Lacerta schreiberi (Squamata: Lacertidae) and Pelodytes spp. (Anura: Pelodytidae), respectively. Cross-species amplification of the squamate markers was evaluated in four genera representing two additional families, whereas for anurans three genera corresponding to three additional families were tested. Three out of the five loci were successfully sequenced in all squamate taxa tested. Cross-amplification of the six anuran markers had lower, but still significant, success. We predict these markers will be of great utility for both population genetics and phylogenetic studies.

High levels of population subdivision in a morphologically conserved Mediterranean toad (Alytes cisternasii) result from recent, multiple refugia: evidence from mtDNA, microsatellites and nuclear genealogies

Pleistocene glaciations often resulted in differentiation of taxa in southern European peninsulas, producing the high levels of endemism characteristic of these regions (e.g. the Iberian Peninsula). Despite their small ranges, endemic species often exhibit high levels of intraspecific differentiation as a result of a complex evolutionary history dominated by successive cycles of fragmentation, expansion and subsequent admixture of populations. Most evidence so far has come from the study of species with an Atlantic distribution in northwestern Iberia, and taxa restricted to Mediterranean‐type habitats remain poorly studied. The Iberian Midwife toad (Alytes cisternasii) is a morphologically conserved species endemic to southwestern and central Iberia and a typical inhabitant of Mediterranean habitats. Applying highly variable genetic markers from both mitochondrial and nuclear genomes to samples collected across the species’ range, we found evidence of high population subdivision within A. cisternasii. Mitochondrial haplotypes and microsatellites show geographically concordant patterns of genetic diversity, suggesting population fragmentation into several refugia during Pleistocene glaciations followed by subsequent events of geographical and demographic expansions with secondary contact. In addition, the absence of variation at the nuclear β‐fibint7 and Ppp3caint4 gene fragments suggests that populations of A. cisternasii have been recurrently affected by episodes of extinction and recolonization, and that documented patterns of population subdivision are the outcome of recent and multiple refugia. We discuss the evolutionary history of the species with particular interest in the increasing relevance of Mediterranean refugia for the survival of genetically differentiated populations during the Pleistocene glaciations as revealed by studies in co‐distributed taxa.

Early pseudoprogression following chemoradiotherapy in glioblastoma patients: The value of RANO evaluation

Introduction. The aim of this study was to determine the frequency of pseudoprogression in a cohort of glioblastoma (GBM) patients following radiotherapy/temozolomide (RT/TMZ) by comparing Macdonald criterial to Response Assessment in Neuro-Oncology (RANO) criteria. The impact on prognosis and survival analysis was also studied. Materials and Methods. All patients receiving RT/TMZ for newly diagnosed GBM from January 2005 to December 2009 were retrospectively evaluated, and demographic, clinical, radiographic, treatment, and survival data were reviewed. Updated RANO criteria were used for the evaluation of the pre-RT and post-RT MRI and compared to classic Macdonald criteria. Survival data was evaluated using the Kaplan-Meier and log-rank analysis. Results and Discussion. 70 patients were available for full radiological response assessment. Early progression was confirmed in 42 patients (60%) according to Macdonald criteria and 15 patients (21%) according to RANO criteria. Pseudoprogression was identified in 10 (23.8%) or 2 (13.3%) patients in Macdonald and RANO groups, respectively. Cumulative survival of pseudoprogression group was higher than that of true progression group and not statistically different from the non-progressive disease group. Conclusion. In this cohort, the frequency of pseudoprogression varied between 13% and 24%, being overdiagnosed by older Macdonald criteria, which emphasizes the importance of RANO criteria and new radiological biomarkers for correct response evaluation.

The prognostic impact of TERT promoter mutations in glioblastomas is modified by the rs2853669 single nucleotide polymorphism

Human hotspot TERT promoter (TERTp) mutations have been reported in a wide range of tumours. Several studies have shown that TERTp mutations are associated with clinicopathological features; in some instances, TERTp mutations were considered as biomarkers of poor prognosis. The rs2853669 SNP, located in the TERT promoter region, was reported to modulate the increased TERT expression levels induced by the recurrent somatic mutations. In this study we aimed to determine the frequency and prognostic value of TERTp mutations and TERT rs2853669 SNP in 504 gliomas from Portuguese and Brazilian patients. TERTp mutations were detected in 47.8% of gliomas (216/452). Glioblastomas (GBM) exhibited the highest frequency of TERTp mutations (66.9%); in this glioma subtype, we found a significant association between TERTp mutations and poor prognosis, regardless of the population. Moreover, in a multivariate analysis, TERTp mutations were the only independent prognostic factor. Our data also showed that the poor prognosis conferred by TERTp mutations was restricted to GBM patients carrying the rs2853669 A allele and not in those carrying the G allele. In conclusion, the presence of TERTp mutations was associated with worse prognosis in GBM patients, although such association depended on the status of the rs2853669 SNP. The status of the rs2853669 SNP should be taken in consideration when assessing the prognostic value of TERTp mutations in GBM patients. TERTp mutations and the rs2853669 SNP can be used in the future as biomarkers of glioma prognosis.

Juvenile psammomatoid ossifying fibroma of the orbit and paranasal sinuses. A case report

Juvenile psammomatoid ossifying fibroma (JPOF) is an uncommon benign fibro-osseous lesion predominantly arising in the paranasal sinuses and orbits of children and young adults. We report a case of JPOF involving the paranasal sinuses and orbit in a 15-year-old boy that presented due to progressive proptosis and downward displacement of the left eye. The lesion, first described as fibrous dysplasia (FD), was totally removed surgically, and then proved to be a JPOF, by histopathology. We discuss its differential diagnosis with other fibro-osseous lesions, histopathological features, and treatment options.

Ossifying Fibromas of the Craniofacial Skeleton

Ossifying fibromas (OF) of the craniofacial skeleton, as described in WHO classification of odontogenic tumors (2005)(Barnes L 2005), are benign fibro-osseous neoplasms characterized by the replacement of normal bone by a fibrous cellular stroma containing foci of mineralized bone trabeculae and cementum-like material that vary in amount and appearance.(Brannon and Fowler 2001; El-Mofty 2002; Cruz, Alencar et al. 2008) The accurate nature and classification of OF has undergone considerable debate among pathologists, resulting in a confusing evolution of competing nomenclatures.(Brannon and Fowler 2001; Sarode, Sarode et al. 2011) Contemporary reviews have classified benign fibroosseous lesions of the craniofacial complex into neoplasms, developmental dysplastic lesions and inflammatory/reactive processes. [Table 1](Eversole, Su et al. 2008) In this reviews, subtypes vary with regard to behavior and propensity for recurrence after surgical excision. The definitive diagnosis can rarely be rendered on the basis of histopathological features alone and is usually dependent upon assessment of microscopic, clinical and imaging features together. This review will discuss the clinical, microscopic, radiological and therapeutic aspects of ossifying fibromas in this localization.

One-step tunneling of DBS extensions--a technical note.

BackgroundInfection constitutes a serious adverse event in deep brain stimulation (DBS) surgery, being responsible for difficult therapeutic decisions that may ultimately involve the removal of implanted material. Some cases begin with skin erosion and wound dehiscence of the retroauricular incision, which is one of the most fragile points. Several techniques of rotation flaps and skin reconstruction, as well as prolonged antibiotic regimens, have been proposed as therapeutic options. To prevent the onset of this complication, the authors propose a one-step tunneling technique of DBS extensions, avoiding the opening of the retroauricular space.MethodsWe describe a surgical technique of a one-step tunneling of DBS extensions in 20 patients submitted to subthalamic DBS for Parkinson’s disease, avoiding the opening of the retroauricular space.ResultsAfter implantation of the extensions using this technique, we had no erosions of the retroauricular skin, with a consequent reduction in the number of infections.ConclusionsThe authors describe an easy surgical technique that allows reduction of wound and erosion complications, with great benefits for DBS patients.

Spinal intramedullary ependymal cyst - Current concepts for diagnosis and surgical management

Abstract We describe a case of a 48-year-old woman who presented with neck pain and progressive left arm weakness. Magnetic Resonance Imaging showed a cervical intramedullary cystic lesion. The lesion was removed and neuropathological analysis revealed an ependymal cyst.

Analysis of a synchronous gliosarcoma and meningioma with long survival: A case report and review of the literature

Background: The simultaneous occurrence of multiple intracranial neoplasms has been reported, especially in genetic familial syndromes and after cranial irradiation. In the absence of these etiologic factors, some reports showed simultaneous occurrence of glioblastoma and meningioma but the association between gliosarcoma and meningioma is unknown. Case Description: We report a case of a 51-year-old woman with synchronous gliosarcoma and meningioma in whom extensive immunohistochemical characterization and molecular profile was performed. The gliosarcoma recurred 21 months after the first resection, reaching 3 years of overall survival. A molecular characterization of all three lesions was performed. None of the lesions showed the presence of mutations in TP53 and BRAF genes. MGMT analysis showed the presence of loss of expression associated with promoter hypermethylation in both gliosarcoma lesions. EGFR overexpression and gene amplification was found only in the recurrent gliosarcoma. Conclusion: The immunohistochemistry and molecular data of this unique case, suggest the distinct clonal origin of meningioma and gliosarcoma lesions, and the association of MGMT methylation with the presumable favorable prognosis observed.

Decompressive craniectomy for massive internal carotid artery infarction after pediatric penetrating neck trauma

Pediatric penetrating carotid arterial trauma is a rare unreported cause of malignant cerebral infarction. Despite increasing evidence of benefit of decompressive hemicraniectomy (DCH) in pediatric malignant stroke, indications and predictors of outcome remain controversial. We report a 4-year-old boy with penetrating zone II neck trauma with laceration of the right internal carotid artery who developed malignant cerebral infarction requiring DCH. Impressive neurological recovery and excellent functional outcome was observed with good psychomotor development and quality of life. To our knowledge, this is the first reported case of pediatric malignant ICA infarction due to penetrating arterial trauma with good neurologic outcome after DCH.