Clinicopathological and molecular analyses of uterine carcinosarcomas using next-generation sequencing: A single-center experience

Background: Uterine carcinosarcomas (UCS) constitute 3–4% of all uterine malignancies and 16% of deaths caused due to uterine neoplasms. Aim: In this study, we aimed to perform DNA-based mutation analysis in 12 genes (KRAS, NRAS, EGFR, C-KIT, BRAF, PDGFRA, ALK, ERBB2, ERBB3, ESR1, RAF1, PIK3CA) to determine the molecular subtypes of UCS using next-generation sequencing (NGS) in patients with aggressive UCS and poor prognosis. We aimed to compare the results of our analysis with clinicopathological data to contribute to the development of targeted therapy approaches related to the molecular changes of UCS. Materials and Methods: In this study, we included 12 cases diagnosed with uterine carcinosarcomas and examined the changes in oncogenes that play a role in UCS pathogenesis. For the analysis of mutation, the clinicopathological data were compared with the variations in the DNA-based gene panel consisting of 12 genes and 1237 variants in the UCS using the NGS method. Results: EGFR mutation was found in 91.7% of the cases, mutation in 41.7%, PDGFRA mutation in 25%, KRAS and PIK3CA mutation in 16.7%, and C-KIT mutation in 8.3% of the cases. Although no statistical significance was found between the detected mutation and clinicopathological data, it was concluded that PDGFRA mutation might be associated with advanced-stage disease development. Conclusion: This study's findings regarding different molecular types of UCS and information on oncogenesis of UCS can provide inferences for targeted therapies in the future by identifying targetable mutations representing early oncogenic events and thereby contribute toward further studies on this subject.

Síndrome de Cohen: reporte de caso y revisión de la literatura / Cohen syndrome: case report and literature review

Resumen Introducción: el Síndrome de Cohen es una enfermedad genética monogénica autosómica recesiva, que se origina a partir de mutaciones en el gen VPS13B (COH1). Se caracteriza por obesidad, retraso psicomotor, microcefalia, hipotonía, miopía progresiva, distrofia retiniana, neutropenia intermitente y rasgos faciales particulares. Objetivo: presentar el segundo caso reportado en Colombia, que fue confirmado mediante estudio molecular. También se presenta una breve revisión de la literatura médica más reciente sobre esta patología. Caso clínico: adolescente de 14 años con microcefalia, trastorno cognitivo, malformaciones menores asociadas, neutropenia y obesidad, con mutación homocigota del gen VPS13B. Conclusión: a pesar de ser un síndrome poco común, con importante variabilidad fenotípica, debe sospecharse con base en los criterios clínicos y en las patologías asociadas.

Abstract Introduction: cohen's syndrome is an autosomal recessive monogenic genetic disease, which originates from mutations in the VPS13B (COH1) gene. It is characterized by obesity, psychomotor retardation, microcephaly, hypotonia, progressive myopia, retinal dystrophy, intermittent neutropenia, and classic facial features. Objective: to present the second case reported in Colombia, which was confirmed by molecular study. A brief review of the most recent medical literature on this pathology is also presented. Clinical case: a 14-year-old adolescent with microcephaly, cognitive disorder, minor associated malformations, neutropenia, and obesity, with a homozygous VPS13B gene mutation. Conclusion: despite being a rare syndrome, with significant phenotypic variability, it should be suspected based on clinical criteria and associated pathologies.

Neoechinorhynchus macrospinosus (Acanthocephala: Neoechinorhynchidae) in Rabbit fish Siganus rivulatus (Siganidae): morphology and phylogeny / Neoechinorhynchus macrospinosus (Acanthocephala: Neoechinorhynchidae) em peixes-coelho Siganus rivulatus (Siganidae): morfologia e filogenia

Siganids are the most important marine fish distributed along the African coast. Therefore, the current study aimed to investigate parasite fauna infects one of the most important mariculture fish species in the Red Sea, the Rabbit fish Siganus rivulatus. One acanthocephalan species has been isolated from the posterior region of fish intestine, belonging to the Neoechinorhynchidae family, and named as Neoechinorhynchus macrospinosus Amin & Nahhas, 1994 based on its morphological and morphometric features. In order to determine the accurate taxonomic position of this acanthocephalan species, molecular phylogenetic analysis was carried out based on the partial sequences of 18S rDNA gene region. The obtained data revealed that this species was associated with a close identity ˃71% for other species belonging to the Neoechinorhynchidae family. In addition, the recovered species deeply embedded in the Neoechinorhynchus genus, closely related to the previously described Neoechinorhynchus sp., N. mexicoensis, and N. golvani with identity percent of 95.14, 93.59, 93.59%, respectively. Therefore, the present study provide a better understanding about the taxonomic status of N. macrospinosus based on 18S rDNA that can be useful for achieving a proper assessment of biodiversity.

Os siganídeos são os peixes marinhos mais importantes distribuídos ao longo da costa africana. Portanto, o presente estudo teve como objetivo investigar a fauna de parasitas infectando uma das espécies mais importantes de peixes para maricultura no Mar Vermelho, o peixe-coelho Siganus rivulatus. Uma espécie de acantocéfalo foi isolada da região posterior do intestino de peixes pertencentes à família Neoechinorhynchidae, e denominadas Neoechinorhynchus macrospinosus Amin & Nahhas, 1994, com base em suas características morfológicas e morfométricas. A fim de determinar a posição taxonômica precisa dessa espécie de acantocéfalo, a análise filogenética molecular foi realizada com base nas sequências parciais da região do gene 18S rDNA e revelou que essa espécie estava associada a uma identidade próxima de até 71% para outras espécies pertencentes a família Neoechinorhynchidae e profundamente enraizada no gênero Neoechinorhynchus, intimamente relacionada a Neoechinorhynchus sp., N. mexicoensis, and N. golvani descrito anteriormente com percentual de identidade de 95,14, 93,59, 93,59%, respectivamente. Portanto, o presente estudo fornece uma melhor compreensão sobre o status taxonômico de N. macrospinosus com base no 18S rDNA que pode ser útil para obter uma avaliação adequada da biodiversidade.

[RETRACTED ARTICLE] Neoechinorhynchus macrospinosus (Acanthocephala: Neoechinorhynchidae) in Rabbit fish Siganus rivulatus (Siganidae): morphology and phylogeny / [ARTIGO RETRATADO] Neoechinorhynchus macrospinosus (Acanthocephala: Neoechinorhynchidae) em peixes-coelho Siganus rivulatus (Siganidae): morfologia e filogenia

Abstract Siganids are the most important marine fish distributed along the African coast. Therefore, the current study aimed to investigate parasite fauna infects one of the most important mariculture fish species in the Red Sea, the Rabbit fish Siganus rivulatus. One acanthocephalan species has been isolated from the posterior region of fish intestine, belonging to the Neoechinorhynchidae family, and named as Neoechinorhynchus macrospinosus Amin & Nahhas, 1994 based on its morphological and morphometric features. In order to determine the accurate taxonomic position of this acanthocephalan species, molecular phylogenetic analysis was carried out based on the partial sequences of 18S rDNA gene region. The obtained data revealed that this species was associated with a close identity ˃71% for other species belonging to the Neoechinorhynchidae family. In addition, the recovered species deeply embedded in the Neoechinorhynchus genus, closely related to the previously described Neoechinorhynchus sp., N. mexicoensis, and N. golvani with identity percent of 95.14, 93.59, 93.59%, respectively. Therefore, the present study provide a better understanding about the taxonomic status of N. macrospinosus based on 18S rDNA that can be useful for achieving a proper assessment of biodiversity.

Resumo Os siganídeos são os peixes marinhos mais importantes distribuídos ao longo da costa africana. Portanto, o presente estudo teve como objetivo investigar a fauna de parasitas infectando uma das espécies mais importantes de peixes para maricultura no Mar Vermelho, o peixe-coelho Siganus rivulatus. Uma espécie de acantocéfalo foi isolada da região posterior do intestino de peixes pertencentes à família Neoechinorhynchidae, e denominadas Neoechinorhynchus macrospinosus Amin & Nahhas, 1994, com base em suas características morfológicas e morfométricas. A fim de determinar a posição taxonômica precisa dessa espécie de acantocéfalo, a análise filogenética molecular foi realizada com base nas sequências parciais da região do gene 18S rDNA e revelou que essa espécie estava associada a uma identidade próxima de até 71% para outras espécies pertencentes a família Neoechinorhynchidae e profundamente enraizada no gênero Neoechinorhynchus, intimamente relacionada a Neoechinorhynchus sp., N. mexicoensis, and N. golvani descrito anteriormente com percentual de identidade de 95,14, 93,59, 93,59%, respectivamente. Portanto, o presente estudo fornece uma melhor compreensão sobre o status taxonômico de N. macrospinosus com base no 18S rDNA que pode ser útil para obter uma avaliação adequada da biodiversidade.

First record of Pseudoterranova decipiens (Nematoda, Anisakidae) infecting the Red spot emperor Lethrinus lentjan in the Red Sea / Primeiro registro de Pseudoterranova decipiens (Nematoda, Anisakidae) infectando o imperador da mancha vermelha Lethrinus lentjan no Mar Vermelho

Abstract The current parasitological study was carried out to investigate helminth parasites infecting the Red spot emperor Lethrinus lentjan inhabiting Hurghada City at the Gulf of Suez, Red Sea, Egypt. Third-stage larvae of nematode parasite was isolated from the intestine as well as body cavity of the examined fish. Light and scanning electron microscopy revealed that this parasite belonged to Anisakidae family within the genus Pseudoterranova. The present species is named Pseudoterranova decipiens based on the presence of triangular mouth aperture with prominent boring teeth and soft swellings of the cuticle, long muscular esophagus, ventrally excretory pore, and narrow transverse slit of anal opening followed by a short mucron. The morphological characteristics of this species were confirmed by molecular analysis of 18S rDNA gene region of the present parasite. It demonstrated a close identity ≥89% with taxa under family Anisakidae, 85% with Raphidascarididae, and 79-84% with Toxocaridae. A preliminary genetic comparison between gene sequence of the present parasite and other oxyurid species placeed it as a putative sister taxon to other Pseudoterranova decipiens described previously. This study demonstrated that the 18S rDNA gene region of Pseudoterranova decipiens yielded a unique sequence that confirmed its taxonomic position in Anisakidae.

Resumo O presente estudo parasitológico foi realizado para investigar os helmintos parasitos que infectam o peixe imperador Lethrinus lentjan, que habita a cidade de Hurghada no Golfo de Suez, Mar Vermelho, no Egito. Larvas de terceiro estágio de parasitos nematoides foram isoladas do intestino e da cavidade do corpo do peixe examinado. Microscopia eletrônica de luz e de varredura revelou que este parasita pertence à família Anisakidae dentro do gênero Pseudoterranova. A espécie atual é denominada Pseudoterranova decipiens baseada na presença de abertura triangular da boca com dentes proeminentes chatos e inchaços moles da cutícula, esôfago muscular longo, poro ventralmente excretor e fenda transversal estreita da abertura anal seguida por um mucron curto. As características morfológicas desta espécie foram confirmadas pela análise molecular da região do gene 18S rDNA do presente parasito. Demonstrou uma identidade próxima ≥89% com taxa sob família Anisakidae, 85% com Raphidascarididae, e 79-84% com Toxocaridae. Uma comparação genética preliminar entre a sequência genética do presente parasito e outras espécies de oxiurídeos coloca-o como um taxon irmão putativo para outros Pseudoterranova descritos anteriormente. Este estudo demonstra que a região do gene 18S rDNA de Pseudoterranova decipiens produz uma sequência única que confirma sua posição taxonômica em Anisakidae.

Neoplasias quísticas pancreáticas: enfoque diagnóstico / Diagnostic Approach to Cystic Pancreatic Neoplasms

Resumen Los quistes pancreáticos comprenden una amplia variedad de lesiones cada vez más frecuentemente diagnosticadas debido tanto al empleo creciente de técnicas de imagen como al envejecimiento de la población. Entre ellas, las neoplasias quísticas mucinosas son especialmente relevantes por su potencial de malignización. Aunque la ecografía abdominal, la tomografía axial computarizada y la resonancia magnética suelen ser las pruebas diagnósticas de imagen iniciales, muchas veces los hallazgos morfológicos no son suficientes para su diferenciación. La ecoendoscopia se ha convertido en la mejor prueba para su caracterización ya que permite realizar estudio morfológico y también del líquido obtenido mediante punción, aunque su precisión diagnóstica para la detección de quistes de estirpe mucinosa y de malignidad sigue siendo baja. La importancia de la adecuada caracterización radica tanto en la detección precoz de las lesiones preneoplásicas y malignas como en evitar cirugías innecesarias. La indicación de ecoendoscopia, de tratamiento quirúrgico y de seguimiento varía entre las distintas guías de práctica clínica estando actualmente en duda especialmente el tratamiento y seguimiento de las neoplasias quísticas mucinosas papilares intraductales de rama lateral por el menor riesgo de degeneración y su asociación con el cáncer de páncreas.

Abstract Cystic pancreatic lesions comprise a wide variety of lesions that are being increasingly diagnosed due to the more frequent use of imaging techniques and the aging of the population. Among these lesions, mucinous cystic neoplasms are especially relevant because of their malignant potential. Although abdominal ultrasound, computerized tomography and magnetic resonance imaging are usually the initial diagnostic imaging tests, morphological findings are often not enough for their differentiation. Endoscopic ultrasound has become the best test for their characterization because it allows morphological study and fluid analysis obtained by puncture of the lesion, although its diagnostic accuracy for the detection of mucinous and malignant cysts remains low. The importance of proper characterization is the early detection of preneoplastic as well as malignant lesions and to avoid unnecessary surgery. Clinical practice guidelines differ about the indications for endoscopic ultrasound, surgical treatment and follow-up of these lesions. Questions specially remains in the management of side-branch intraductal papillary neoplasm because of their lower risk of degeneration and their association with pancreatic cancer.

Genetic and molecular aspects of androgenetic alopecia

Androgenetic alopecia is the most common form of progressive hair loss in humans. A genetic predisposition and hormonal status are considered as major risk factors for this condition. Several recent advances in molecular biology and genetics have increased our understanding of the mechanisms of hair loss in androgenetic alopecia. We review these advances and examine the trends in the genetic and molecular aspects of androgenetic alopecia.

Occurrence of Brown Blight Caused by Waitea circinata var. zeae on Cool Season Turfgrass in Korea

In 2010, disease symptoms, including necrotic lesions on stems and leaves with circular yellow-brown or irregular brown color patches, were observed on cool-season turfgrass at golf courses (OHCC) and the Daegu University research farm in Gyeongbuk, Korea. We isolated the causal agent and identified it as Waitea circinata var. zeae by morphological characterization and molecular analysis. To the best of our knowledge, this is the first report of brown patch caused by W. circinata var. zeae on cool-season turfgrass in Korea.

Mycological and Molecular Analysis of Soil Keratinophilic Fungi Isolated in Summer Beaches of the Korean South Coast / 대한피부과학회지

BACKGROUND: Some species of soil keratinophilic fungi (KPF) are known to be pathogens that may lead to cutaneous infection. People exposed to these species through direct contact with soil on beaches can contract KPF infection. However, there is little literature regarding pathogenic KPF isolated from beaches during summer time. OBJECTIVE: This study aims to evaluate the distribution of soil KPF isolated from beaches in Korea during summer. METHODS: One hundred eighty soil samples from six beaches in the southern coastline of Korea under three different climatic conditions were collected. The KPF species were isolated using the hair-baiting technique. Then, molecular identification was performed by sequencing the ribosomal RNA (rRNA) internal transcribed spacer (ITS) region to investigate the exact species of the isolated fungi. RESULTS: Among the one hundred eighty soil samples, twenty-nine strains (16.1%) of KPF were recovered. The isolation rate of KPF among the beaches varied from 0 to 34.5%. KPF was most frequently isolated in shaded dry areas (30%), followed by sunny dry areas (18.3%), and sunny wet areas (0%). Molecular identification of the fungi using rRNA ITS analysis helped in their classification. Microsporum gypseum/Arthroderma incurvatum (69.0%), Microsporum gypseum/Arthroderma gypseum (3.4%), Trichophyton ajelloi/Arthroderma uncinatum (13.8%), Microsporum cookei/Arthroderma cajetani (10.3%), and Chrysosporium indicum/Aphanoascus terreus (3.4%) were identified. Single nucleotide polymorphism (SNP) was observed at position 180 of the rRNA ITS2 in the 20 strains of Microsporum gypseum/Arthroderma incurvatum, and the species was divided into Types 1 (14 strains) and 2 (6 strains) depending on the base present at the SNP position. The geographic distribution of these two types differed. CONCLUSION: Our results show that the beach is a possible source of keratinophilic fungal infection in humans. People should be aware of pathogenic fungi on the soil of beaches during summer and take measures to prevent possible superficial fungal infections.

Beta talasemia intermedia: características clínicas y estudio molecular. Serie de casos clínicos / Beta thalassemia intermedia: clinical characteristics and molecular analysis. Case series

La beta talasemia intermedia es una hemoglobinopatía de amplio espectro clínico, que surge de la presencia de una o dos mutaciones en el gen HBB, asociada a modificadores genéticos secundarios y/o terciarios. Analizamos las características clínicas y de laboratorio de 29 pacientes con beta talasemia intermedia, evaluados en un período de 23 años. La edad mediana fue de 10,8 años (rango: 0,34-60,4). El 100% de los pacientes mostró anemia microcítica hipocrómica, y solo el 17,2% presentó esplenomegalia y requerimiento transfusional esporádico. El análisis molecular de los pacientes detectó 3 con los dos genes HBB afectados; 2 con un gen HBB afectado y genes alfa cuadriplicados/triplicados; 23 con un gen HBB afectado y genes alfα triplicados; y 1 con dos genes HBB afectados y polimorfismos de genes gama. La correcta identificación de estos pacientes aseguró un adecuado consejo genético y la implementación de controles clínicos regulares.

Beta thalassemiaintermediaisaquantitative haemoglobinopathy covering a broad clinical spectrum, that results from the presence of one or two HBB gene mutations associated with secondary and/or tertiary genetic modifiers. We analyze the clinical and laboratory features of 29 patients with beta thalassemia intermedia, assessed over a period of 23 years. Median age was 10.8 years (range: 0.34-60.4). Hypochromic microcytic anemia was seen in 100% of the patients, while only 17.2% had splenomegaly and occasional transfusion requirement. The molecular analysis of patients detected: 3 with two HBB affected genes; 2 with one HBB affected gene and alpha quadruplicate/triplicate genes; 23 with one HBBaffected gene and alpha triplicate genes and 1 with two HBB affected genes and polymorphisms of gamma genes. The adequate identification of these patients enables us to give appropriate genetic counseling and implementation of regular clinical follow up

Implicações de estratégias molares/moleculares na análise comportamental da cultura / Implications of molar/molecular strategies for behavioral analysis of culture / Implicaciones de estrategias molares/moleculares en análisis del comportamiento de la cultura

Desde as contribuições seminais de Glenn, a compreensão de fenômenos socioculturais na perspectiva da Análise do Comportamento tem sido fortemente influenciada pela premissa da necessidade de um novo nível de análise. Esta premissa tem favorecido análises molares dos fenômenos socioculturais, cuja ênfase recai sobre o grupo ou as unidades sociais e culturas identificadas como objeto de seleção. No entanto, por meio de análises que incidirão principalmente sobre os conceitos e exemplos propostos por Glenn, argumentamos que, em contrapartida, tais análises molares podem obscurecer variáveis relevantes do ponto de vista molecular. Além disso, avaliamos as implicações em termos de estratégias de investigação dos fenômenos socioculturais que a adoção quase exclusiva de análises molares tem tido para a construção de uma teoria comportamentalista da cultura.

Since the seminal contributions of Glenn, analysis of sociocultural phenomena in Behavior Analysis has been strongly influenced by assumption of need of a new analysis level. This assumption has favored a molar perspective towards sociocultural phenomena, whose emphasis is on group or social/cultural units identified as object of selection. However, through analyzes that focus on concepts and examples proposed by Glenn, we argue that, on the other hand, those analyzes obscure some relevant variables from the standpoint of a molecular perspective. Furthermore, this article discussed implications that adoption almost exclusive of molar perspectives have had to construction of a behavioral theory of culture.

Desde las contribuciones de Glenn, la comprensión de los fenómenos socio-culturales en Análisis del Comportamiento ha sido fuertemente influenciada por la premisa de la necesidad de un nuevo nível de análisis. Esta premisa ha favorecido el análisis molar de los fenómenos socio-culturales, cuyo énfasis está en el grupo o unidades sociales y culturales identificadas como un objeto de selección. Sin embargo, a través del análisis que se centrará principalmente en los conceptos y ejemplos propuestos por Glenn, argumentamos que, em cambio, estos análisis molares pueden oscurecer variables importantes del punto de vista molecular. Además, evaluamos las implicaciones en términos de estrategias de investigación de los fenómenos socioculturares que la adopción casi exclusiva de perspectivas molares ha tenido para la construcción de una teoría comportamental de la cultura.

Fast and robust protocol for prenatal diagnosis of mucopolysaccharidosis type II

Introduction: Mucopolysaccharidosis type II (MPSII) is an X-linked lysosomal disorder caused by deficiency of iduronate-2-sulfatase (IDS). In this study, we proposed a new protocol for prenatal diagnosis, using DNA obtained from amniotic fluid cells that did not attach to the bottom of the culture flask after the first medium change. Methods: Four pregnant MPS II carriers were referred to the Medical Genetics Service of Hospital de Clinicas de Porto Alegre for a prenatal diagnosis and identification of the disease, which were performed by polymerase chain reaction (PCR) amplification, restriction fragment length polymorphism, and sequencing according to the mutation previously found in the family. Results: The analysis indicated the absence of mutation in three fetal materials and the presence of mutation in one case. Concomitantly, cytogenetic and biochemical analyses were performed after 12 days of cell culture, and only one case showed absence of enzyme activity, confirming the molecular analysis. Conclusions: This diagnostic protocol designed to provide more robust results and safer genetic counseling suggests that DNA obtained from floating amniotic fluid cells can be used as a source of fetal material to allow a faster alternative for prenatal care through molecular analysis. Determination of IDS gene mutation in fetal amniotic fluid cells together with IDS enzyme activity testing is a rapid, sensitive and accurate method for prenatal diagnosis of MPS II for high-risk pregnant women.

Molecular characteristic of alpha thalassaemia among patients diagnosed in UKM Medical Centre

Alpha (α) thalassaemia is the most common inherited disorder in Malaysia. The clinical severity is dependant on the number of α genes involved. Full blood count (FBC) and haemoglobin (Hb) analysis using either gel electrophoresis, high performance liquid chromatography (HPLC) or capillary zone electrophoresis (CE) are unable to detect definitively alpha thalassaemia carriers. Definitive diagnosis of α-thalassaemias requires molecular analysis and methods of detecting both common deletional and non-deletional molecular abnormailities are easily performed in any laboratory involved in molecular diagnostics. We carried out a retrospective analysis of 1623 cases referred to our laboratory in Universiti Kebangsaan Malaysia Medical Centre (UKMMC) for the diagnosis of α-thalassaemia during the period October 2001 to December 2012. We examined the frequency of different types of alpha gene abnormalities and their haematologic features. Molecular diagnosis was made using a combination of multiplex polymerase reaction (PCR) and real time PCR to detect deletional and non-deletional alpha genes relevant to southeast Asian population. Genetic analysis confirmed the diagnosis of α-thalassaemias in 736 cases. Majority of the cases were Chinese (53.1%) followed by Malays (44.2%), and Indians (2.7%). The most common gene abnormality was αα/--SEA (64.0%) followed by αα/-α3.7 (19.8%), -α3.7 /--SEA (6.9%), αα/ααCS (3.0%), --SEA/--SEA (1.2%), -α3.7/-α3.7 (1.1%), αα/-α4.2 (0.7%), -α4.2/--SEA (0.7%), -α3.7/-α4.2 (0.5%), ααCS/-- SEA (0.4%), ααCS/ααCd59 (0.4%), ααCS/ααCS (0.4%), -α3.7/ααCd59 (0.3%), αα/ααCd59 (0.1%), αα Cd59/ ααIVS I-1 (0.1%), -α3.7/ααCS (0.1%) and --SEA /ααCd59 (0.1%). This data indicates that the molecular abnormalities of α-thalassaemia in the Malaysian population is heterogenous. Although α-gene deletion is the most common cause, non-deletional α-gene abnormalities are not uncommon and at least 3 different mutations exist. Establishment of rapid and easy molecular techniques is important for definitive diagnosis of alpha thalassaemia, an important prerequisite for genetic counselling to prevent its deleterious complications.

Isolation,identification and full-length genome sequence of EMCV f rom domesticated boar / 中国人兽共患病学报

Encephalomyocarditis virus (EMCV) ,named JZ1202 ,was isolated from domesticated boar in Henan ,China . We performed the full-length genome sequencing and molecular characteristic analysis of the isolated strain .Results showed that the full-genome sequence of EMCV JZ1202 generated a sequence of 7 735 bp in length ,and had 81 .2%-99 .9% nucleotide identity with other reference strains from different animals ,but 99 .4% with Chinese reference from pig .The phylogenetic tree was constructed based on the full-length genome;ORF and VP1 gene sequences identified EMCV was divided into G1 ,G2 and G3 groups ;the strain JZ1202 belongs to G1 with other Chinese reference strains .Results identified that the EMCV infection could cause severe clinical symptoms in domesticated boar .Big regional differences exist in EMCV and the transmission is limit-ed in a range of area .However ,cross infection and prevalence of EMCV disease between domesticated boar and mice might ex-ist .Mutation of some amino acid may occurred in EMCV infected domesticated boar .

Dumb-bell shaped poorly differentiated pelvic synovial sarcoma with molecular confirmation: A rare presentation of an uncommon disease entity.

Pelvic localization of synovial sarcoma is a rare phenomenon and to the best of our knowledge its presentation as a large "dumb-bell"-shaped abdomino-pelvic mass showing extension to the thigh has never been reported in the literature. We report a case of a young adult presenting with retention of urine and was found to have a large abdomino-pelvic mass causing bony destruction and compression of pelvic viscera. A biopsy revealed a cellular tumor composed of spindle to oval cells arranged in a hemangiopericytomatous pattern. Histopathology was suggestive of poorly differentiated synovial sarcoma. Immunohistochemistry (IHC) was positive for vimentin, CD 99, Bcl2, Mic2 and focally for EMA and negative for CD 34, CK, desmin, synaptophysin, and WT1. Due to equivocal IHC findings molecular analysis was done which confirmed the diagnosis as synovial sarcoma.

Comparison of methods for the detection of biofilm formation by Staphylococcus aureus isolated from bovine subclinical mastitis

Biofilm formation is considered to be a selective advantage for Staphylococcus aureus mastitis isolates by facilitating bacterial persistence in the udder. It requires attachment to mammary epithelium, proliferation and accumulation of cells in multilayers. The objective of this study was to determine the sensitivity and specificity of three techniques for the detection of S. aureus biofilm-positive strains. Two phenotypic tests, including growth on microtitre plates and Congo red agar, were compared with a PCR technique using 94 S. aureus strains obtained from cows with subclinical mastitis from two farms in the state of São Paulo. These strains were characterised by in vitro slime production on Congo red agar, biofilm formation on microtitre plates and the presence of the icaA and icaD genes. The results revealed that 85% of the isolates tested produced slime on the Congo red agar, 98.9% of the isolates produced biofilms in vitro by adhering to sterile 96-well "U" bottom polystyrene tissue culture plates, and 95.7% of the isolates carried the icaA and icaD genes. The results of the phenotypic tests for biofilm formation were compared with those of the molecular analysis, and the sensitivity and specificity of the Congo red agar test were 88.9% and 100%, respectively, while those of the microtitre plate test were 100% and 25%, respectively. When the phenotypic methods for the detection of biofilm producers, namely growth on microtitre plates and Congo red agar, were compared, the sensitivity and specificity were 86% and 100%, respectively. Therefore, growth on Congo red agar and the microtitre plate test are methods that could be used to determine whether an isolate has the potential for biofilm production.

Molecular basis of high-level ciprofloxacin resistance in Neisseria gonorrhoeae strains from Shandong Province, China

In the study, the ciprofloxacin resistance rate was 100%. High-level ciprofloxacin resistance rate was 63.55%. Sixteen different mutation patterns involved in the formation of ciprofloxacin resistance were identified. The most prevalent were patterns P7 (25.2%), P8 (15.0%), P9 (11.2%), P1 (10.3%), and P5 (10.3%). All of the 107 NG isolates analyzed for mutations in the study have demonstrated a change of Ser-91 → Phe in the gyrA gene, and all except one have demonstrated a change in position 95 of the amino acid sequence. All of the 68 high-level QRNG isolates had double mutations in gyrA gene combined with a single or two mutations in parC gene. It is most important that a new mutation site of Ile-97 → Met in gyrA and a new mutation of Leu-106 → Ile in parC were found in the study, both leading to high-level ciprofloxacin resistance (MIC values, 8 µg/mL, 32 µg/mL, respectively). Therefore, we confim that gyrA mutations are necessary for the fluoroquinolone resistance phenotype and parC mutations are correlated intimately with high-level fluoroquinolone resistance. In China fluoroquinolone resistance in Neisseria gonorrhoeae strains is very serious and the new mutation sites in the fluoroquinolone resistance-determining regions emerge more and more quickly. Hence, in China fluoroquinolones, which are used to treat gonorrhoea presently, should be substituted by a new antibiotics.

In vitro growth of Ganoderma boninense isolates on novel palm extract medium and virulence on oil palm (Elaeis guineensis) seedlings

Aims: Basal stem rot (BSR) disease caused by Ganoderma pathogenic fungi, especially Ganoderma boninense is thriving rapidly in both areas with coastal and inland soils. The objectives of this study were to isolate and characterize Ganoderma isolates collected from various locations in Peninsular Malaysia through the comparison of their growth rate in vitroly on conventional and novel palm extract media, and to determine the degree of virulence caused by the isolates in oil palm seedlings. Methodology and results: In this study, 12 Ganoderma isolates were collected from infected oil palm trees, from various locations – Johor, Negeri Sembilan, Kedah, Perak, Pahang, and Kelantan, in Malaysia in year 2011. Twelve Ganoderma isolates were identified using molecular method with primer set that targeted at small-subunit 18S rDNA fragment, and characterized by determining the in vitro growth rate, and degree of virulence in 2-month-old oil palm seedlings in the nursery using both disease incidence (DI) and disease severity index (DSI) as the measurements to quantify the infection. All the Ganoderma isolates were identified as G. boninense and sequences of the respective isolates were deposited in GenBank. In general, all the isolates proliferated faster on oil palm extract medium (OPEM) compared to malt extract agar (MEA). Twelve G. boninense isolates were observed to illustrate different degree of virulence ranging from highly pathogenic to least pathogenic. Conclusion, significance and impact of study: Cultures of 12 G. boninense isolates were observed to show faster growth rate (P < 0.014) on OPEM under in vitro conditions compared to conventional MEA medium, except Bt Lintang G10 and GBA G12 isolates. OPEM medium could provide a better alternative for maintaining and culturing Ganoderma strains. In the current study, both DI and DSI were highly correlated. However, there were low linear relationships (R2 < 0.423) between mycelia growth rate (on MEA and OPEM) and degree of virulence (DI and DSI) at 12-, 14- and 16- weeks after treatments among the G. boninense isolates tested. Furthermore, different degrees of virulence in twelve separate Ganoderma isolates were reported. Therefore, it is crucial to incorporate more than one isolate into any researches on screening for Ganoderma resistance or tolerance planting materials, searching for potential biological control agents, and studying bitrophic or tri-trophic interactions. In addition, this study was aimed to isolate G. boninense strains with various virulence levels for future studies.

Immunohistochemical validation of TLE1, a novel marker, for synovial sarcomas.

Background & objectives: Logistic and financial constraints limit application of several available immunohistochemical (IHC) markers and molecular analysis in every case of synovial sarcoma, diagnosed in our settings. Recently, TLE1 has been recognized as a robust IHC marker for diagnosing a synovial sarcoma. Here, we present IHC features of synovial sarcomas, including TLE1 expression in these cases and in some other tumours. Methods: Conventional sections from 42 synovial sarcomas (30 retrospective & 12 prospectively diagnosed) were subjected to TLE1 IHC staining, including 21 tumours confirmed with molecular testing. TLE1 immunostaining was graded from 0, 1+, 2+, 3+, with 2+ or 3+ grades interpreted as positive staining. Results: Of the 42 tumours, 26 (61.9%) were of monophasic spindle cell type, 13 biphasic type (30.9%), two (4.7%) calcifying type and remaining one (2.3%) was a poorly differentiated synovial sarcoma. On immunohistochemistry (IHC), tumours were positive for epithelial membrane antigen (EMA) (26/34, 76.4%), cytokeratin (CK)7 (6/10, 60%), CK/MNF116 (6/21, 28.6%), B cell lymphoma 2 (BCL2) (36/37, 97.3%), cluster of differentiation molecule 99 (MIC2) (23/31, 74.1%) and transducin-like enhancer of split 1 (TLE1) (40/42, 95.2%), while negative for CD34 in all 21 tumours, wherever performed. TLE1 was also positive in tumour controls, including schwannomas (5/5, 100%), neurofibromas (2/2, 100%), malignant peripheral nerve sheath tumors (2/12, 17%) and Ewing sarcomas (4/10, 40%). TLE1 sensitivity for diagnosis of synovial sarcomas was 95.2 per cent. Its overall specificity was 63.7 per cent, whereas with regards to tumors forming its closest differential diagnoses, its specificity was 72 per cent. Interpretation & conclusions: Although molecular confirmation is the diagnostic gold standard for synovial sarcoma, TLE1, in view of its high sensitivity may be a useful marker within the optimal IHC panel comprising EMA, BCL2, MIC2, CD34 and CK7, especially on small biopsy samples, for substantiating a diagnosis of synovial sarcoma. Awareness of TLE1 expression in other tumours and its correct interpretation are necessary.

Coxiella burnetii, the agent of Q fever in Brazil: its hidden role in seronegative arthritis and the importance of molecular diagnosis based on the repetitive element IS1111 associated with the transposase gene

Coxiella burnetii is the agent of Q fever , an emergent worldwide zoonosis of wide clinical spectrum. Although C. burnetii infection is typically associated with acute infection, atypical pneumonia and flu-like symptoms, endocarditis, osteoarticular manifestations and severe disease are possible, especially when the patient has a suppressed immune system; however, these severe complications are typically neglected. This study reports the sequencing of the repetitive element IS1111 of the transposase gene of C. burnetii from blood and bronchoalveolar lavage (BAL) samples from a patient with severe pneumonia following methotrexate therapy, resulting in the molecular diagnosis of Q fever in a patient who had been diagnosed with active seronegative polyarthritis two years earlier. To the best of our knowledge, this represents the first documented case of the isolation of C. burnetii DNA from a BAL sample.