Vitellogenin genes are transcribed in Culex quinquefasciatus ovary

BACKGROUND Culex quinquefasciatus, a cosmopolitan, domestic, and highly anthropophilic mosquito, is a vector of pathogenic arboviruses such as West Nile virus and Rift Valley virus, as well as lymphatic filariasis. The current knowledge on its reproductive physiology regarding vitellogenin expression in different tissues is still limited. OBJECTIVES In this study, we analysed the transcriptional profiles of vitellogenin genes in the fat body and ovaries of C. quinquefasciatus females during the first gonotrophic cycle. METHODS C. quinquefasciatus ovaries and/or fat bodies were dissected in different times during the first gonotrophic cycle and total RNA was extracted and used for reverse transcription polymerase chain reaction, quantitative real time-PCR, and in situ hybridisation. FINDINGS We confirmed the classical descriptions of the vitellogenic process in mosquitoes by verifying that vitellogenin genes are transcribed in the fat bodies of C. quinquefasciatus females. Using RNA in situ hybridisation approach, we showed that vitellogenin genes are also transcribed in developing ovaries, specifically by the follicle cells. MAIN CONCLUSIONS This is the first time that vitellogenin transcripts are observed in mosquito ovaries. Studies to determine if Vg transcripts are translated into proteins and their contribution to the reproductive success of the mosquito need to be further investigated.

Understanding professional stratification and hybridisation in the medical profession: exploratory evidence from Brazilian federal university hospitals / Comprendiendo la estratificación e hibridación profesional en la profesión médica: evidencia exploratoria de los hospitales universitarios federales de Brasil / Compreendendo a estratificação profissional e hibridização na profissão médica: evidências exploratórias de hospitais universitários federais brasileiros

Abstract This paper explores drivers, implications, and trends of professional stratification and hybridisation in the medical profession employed in Brazilian substituir por: federal university hospitals (HUFs). Drawing on exploratory findings, we examine some repercussions of the migration of university hospitals to EBSERH, a public company established by the federal government to manage and organise HUFs integrated into the Unified Health System (SUS). Our research shows that transferring hospital administration to EBSERH has led to further internal stratification of the medical workforce. The shift from the logic of medical-academic professionalism to the new logic of business-like healthcare, with the adoption of distinct job contracts and more managerial logics of work and control, may well be changing subjective and formal links established between professionals, universities, and hospitals. We identify and discuss trends towards hybridisation and dehybridisation. These findings are relevant because this shift can have profound implications for the academic nature of HUFs and for the future of professionalism within these health and teaching organisations.

Resumen Este artículo explora impulsores, implicaciones y tendencias de la estratificación y la hibridación profesional en la profesión médica empleada en los hospitales universitarios federales brasileños (HUFs). A partir de hallazgos exploratorios, examinamos algunas de las repercusiones de la migración de los hospitales universitarios a la EBSERH, una empresa pública creada por el gobierno federal para administrar y organizar los HUFs integrados en el Sistema Único de Salud (SUS). Nuestra investigación muestra que la transferencia de la administración hospitalaria a la EBSERH ha llevado a una mayor estratificación interna de la fuerza laboral médica. El cambio de la lógica del profesionalismo médico-académico a una lógica empresarial en salud, con la adopción de diferentes regímenes de empleo y modos de trabajo y control más gerenciales, puede estar alterando los vínculos subjetivos y formales que se establecen entre los profesionales, las universidades y los hospitales involucrados. Se identifican y discuten tendencias hacia la hibridación y deshibridación. Estos hallazgos son relevantes porque tales cambios pueden tener profundas implicaciones para la naturaleza académica de los HUFs, así como para el futuro del profesionalismo dentro de estas organizaciones de salud y educación.

Resumo: Este artigo explora motivadores, implicações e tendências de estratificação e hibridização profissional na profissão médica atuante em hospitais universitários federais brasileiros (HUFs). Com base em achados exploratórios, examinamos algumas das repercussões da migração dos hospitais universitários para a EBSERH, uma empresa pública criada pelo governo federal para gerenciar e organizar os HUFs vinculados ao Sistema Único de Saúde (SUS). A pesquisa mostra que a transferência da administração hospitalar para a EBSERH tem acarretado maior estratificação interna da força de trabalho médica. A mudança da lógica de profissionalismo médico-acadêmico para uma nova lógica empresarial de saúde, com a adoção de distintos regimes empregatícios e formas de trabalho e controle mais gerenciais, podem estar alterando vínculos subjetivos e formais estabelecidos entre os profissionais, as universidades e os hospitais envolvidos. Tendências em direção à hibridização e desibridização foram identificadas e discutidas. Esses achados são relevantes tendo em vista que tais movimentos podem representar implicações profundas para a natureza acadêmica dos HUFs e para o futuro do profissionalismo nessas organizações de saúde e ensino.

FISHing for 1p19q codel in oligodendroglioma

@#Together with isocitrate dehydrogenase (IDH) mutation, co-deletion of 1p19q (1p19q codel) is a prerequisite for diagnosis of oligodendroglioma, making it imperative that histopathology laboratories introduce testing for 1p19q codel. To date there is still no consensus reference range and cut-offs that confirm deletion of 1p or 19q. We embarked on determining our reference range in 11 formalinfixed, paraffin-embedded non-neoplastic brain tissue using fluorescence in situ hybridisation (FISH) with the Vysis 1p36/1q25 and 19q13/19p13 FISH Probe Kit (Abbott Molecular Inc., USA). At same time we attempted to validate our methodology in 13 histologically-confirmed IDH-mutant oligodendrogliomas. For 1p, percentage cells with deletion (range=8-23%; mean±SD = 15.73±5.50%) and target: control (1p36:1q25) ratio (range = 0.89-0.96; mean±SD = 0.92±0.03) in non-neoplastic brain, differed significantly (p<0.000) from oligodendroglioma (percentage cells with deletion: range = 49-100%; mean±SD = 82.46±15.21%; target:control ratio range:0.50-0.76; mean±SD = 0.59±0.08). For 19q, percentage cells with deletion (range = 7-20%; mean±SD = 12.00±3.49%) and target:control (19q13/19p13) ratio (range:0.90-0.97; mean±SD = 0.94±0.02) in non-neoplastic brain also differed significantly from oligodendroglioma (percentage cells with deletion: range = 45-100%; mean±SD = 82.62±18.13%; target:control ratio range:0.50-0.78; mean±SD = 0.59±0.09). Using recommended calculation method, for diagnosis of 1p deletion, percentage of cells showing deletion should be >32-33% and/or target:control ratio <0.83. For 19q, percentage of cells showing deletion should be >22% and target:control ratio <0.88. Using these cut-offs all 13 oligodendroglioma demonstrated 1p19q codel.

Early relapse after complete remission of primary plasma cell leukaemia manifesting clonal evolution: A case report

@#Introduction: Plasma cell leukaemia (PCL) is a rare variant of multiple myeloma. We report a case of PCL to demonstrate the clonal evolution, resulting in disease relapse after achieving complete remission, and its aggressive nature of the disease, leading to poor clinical outcome. Case Report: A 69-year-old man presented with a three-day-history of worsening generalized body weakness, poor oral intake, nausea, significant loss of weight and lower back pain. He was diagnosed as primary PCL, based on hypercalcaemia, renal insufficiency, anaemia, thrombocytopenia, lytic bone lesions, 24% abnormal plasma cells in peripheral blood, immunophenotype of clonal plasma cells which were positive for CD38, CD138 and CD56 markers with kappa light chain restriction, 49% abnormal plasma cells in bone marrow, monoclonal paraprotein (IgG kappa) in serum and urine, and positive IGH rearrangement (Fluorescence in-situ hybridisation, FISH). He achieved complete remission after four cycles of Bortezomib-based therapy. There was a plan for high-dose therapy plus autologous haematopoietic cell transplantation. A month later, the disease relapsed, as evidenced by 94% abnormal plasma cells in his bone marrow aspirate, complex karyotype and abnormal FISH results. He passed away a few days later, from severe septicaemia. Time-to-progression of disease was 1 month and overall survival was 5 months. Discussion: This case report illustrates the clonal evolution and aggressive nature of primary PCL with older age at presentation, leading to a shorter duration of remission and overall survival.

Mycobacterium tuberculosis promotes genomic instability in macrophages

BACKGROUND Mycobacterium tuberculosis is an intracellular pathogen, which may either block cellular defensive mechanisms and survive inside the host cell or induce cell death. Several studies are still exploring the mechanisms involved in these processes. OBJECTIVES To evaluate the genomic instability of M. tuberculosis-infected macrophages and compare it with that of uninfected macrophages. METHODS We analysed the possible variations in the genomic instability of Mycobacterium-infected macrophages using the DNA breakage detection fluorescence in situ hybridisation (DBD-FISH) technique with a whole human genome DNA probe. FINDINGS Quantitative image analyses showed a significant increase in DNA damage in infected macrophages as compared with uninfected cells. DNA breaks were localised in nuclear membrane blebs, as confirmed with DNA fragmentation assay. Furthermore, a significant increase in micronuclei and nuclear abnormalities were observed in infected macrophages versus uninfected cells. MAIN CONCLUSIONS Genomic instability occurs during mycobacterial infection and these data may be seminal for future research on host cell DNA damage in M. tuberculosis infection.

Epstein–Barr virus infection in B-cell Non-Hodgkin’s Lymphomas of the Oral and Maxillofacial Region: Is there any evidence?

Introduction: Epstein–Barr virus (EBV) might be an aetiological agent involved in the pathogenesis of certain Non-Hodgkin’s Lymphomas (NHLs). EBV infection has been diagnosed by serologic testing within the tumour biopsies of patients with NHL. However, the association between EBV and NHL is inconsistent with a preference for certain anatomic sites, histologic subtypes and immunosuppressed patients. The objective of this study was to characterise the B-cell NHLs of the oral cavity and maxillofacial region using histological and immunophenotypical techniques and to determine its association with EBV infection. Materials and Methods: This was a descriptive cross-sectional study that included 14 cases of B-cell NHLs of the oral cavity and maxillofacial region. The haematopoietic and lymphoid tissue tumours classification of WHO was used to categorize the cases. In-situ hybridisation for EBV–encoded RNA was performed to confirm the EBV infection. Results: The average age of the patients included in the study was found to be 48.8 ± 23 years with a higher female to male ratio (1.3:1). Our study suggested that diffuse large B-cell lymphomas (DLBCLs) and Burkitt’s lymphomas (BLs) constitute the predominant subtypes of lymphomas affecting the oral cavity and maxillofacial regions. Conclusion: The findings from our study support the view that at least a relatively smaller proportion of B-cell NHLs that occur in the oral cavity and maxillofacial region do not have a pathogenic association with EBV.

Use of fluorescent oligonucleotide probes for differentiation between Paracoccidioides brasiliensis and Paracoccidioides lutzii in yeast and mycelial phase

BACKGROUND Fluorescence in situ hybridisation (FISH) associated with Tyramide Signal Amplification (TSA) using oligonucleotides labeled with non-radioactive fluorophores is a promising technique for detection and differentiation of fungal species in environmental or clinical samples, being suitable for microorganisms which are difficult or even impossible to culture. OBJECTIVE In this study, we aimed to standardise an in situ hybridisation technique for the differentiation between the pathogenic species Paracoccidioides brasiliensis and Paracoccidioides lutzii, by using species-specific DNA probes targeting the internal transcribed spacer-1 (ITS-1) of the rRNA gene. METHODS Yeast and mycelial phase of each Paracoccidioides species, were tested by two different detection/differentiation techniques: TSA-FISH for P. brasiliensis with HRP (Horseradish Peroxidase) linked to the probe 5’ end; and FISH for P. lutzii with the fluorophore TEXAS RED-X® also linked to the probe 5’ end. After testing different protocols, the optimised procedure for both techniques was accomplished without cross-positivity with other pathogenic fungi. FINDINGS The in silico and in vitro tests show no reaction with controls, like Candida and Cryptococcus (in silico) and Histoplasma capsulatum and Aspergillus spp. (in vitro). For both phases (mycelial and yeast) the in situ hybridisation showed dots of hybridisation, with no cross-reaction between them, with a lower signal for Texas Red probe than HRP-TSA probe. The dots of hybridisation was confirmed with genetic material marked with 4’,6-diamidino-2-phenylindole (DAPI), visualised in a different filter (WU) on fluorescent microscopic. MAIN CONCLUSION Our results indicated that TSA-FISH and/or FISH are suitable for in situ detection and differentiation of Paracoccidioides species. This approach has the potential for future application in clinical samples for the improvement of paracoccidioidomycosis patients prognosis.

Performance of a molecular assay to detect Mycobacterium tuberculosis complex DNA in clinical specimens: multicenter study in Brazil

BACKGROUND In high tuberculosis (TB) burden countries, there are few data on the performance of new molecular commercialised assays developed locally. OBJECTIVE To evaluate the performance of a new molecular commercialised assay for TB diagnosis (Detect-TB) in three laboratories. METHODS A total of 302 sputum samples from an equal number of patients with presumptive diagnosis of pulmonary tuberculosis (PTB) were submitted for routine smear microscopy, culture, and Detect-TB assay at three different sites in Brazil (the cities of Caxias do Sul, São Paulo and Canoas). FINDINGS Seventy four (24.7%) TB cases were diagnosed (65 bacteriologically confirmed). When compared to smear microscopy/culture results, the overall sensitivity and specificity of Detect-TB assay was 84.6% (CI 95%; 73.7-91.6) and 93.1% (CI 95%; 89.1-95.8), respectively. When compared to bacteriological and clinical diagnostic criteria, the sensitivity and specificity of Detect-TB assay was 74.3% (CI 95%; 63.3-82.9) and 92.9% (CI 95%; 88.7-95.6), respectively. Among the three sites - Caxias do Sul, São Paulo and Canoas - the sensitivity and specificity were respectively 94.7% and 97.8%; 71.4% and 93.9%, 82.1% and 88.9%. MAIN CONCLUSIONS These findings suggest that the Detect-TB assay could be applied routinely in reference laboratories across different regions in Brazil.

Síndrome de Sotos diagnosticado por hibridación genómica comparativa / Sotos syndrome diagnosed by comparative genomic hybridisation

El síndrome de Sotos (SS) es una enfermedad genética con un patrón de herencia autosómico dominante, causado por haploinsuficiencia del gen NSD1 secundaria a mutaciones puntuales o microdeleciones del locus 5q35 en el que está ubicado el gen. Es un síndrome poco frecuente, presentándose en 7 de cada 100.000 nacimientos. El objetivo de este reporte es presentar el caso de una paciente de 4 años con retardo global del desarrollo, y hallazgos físicos especiales que sugerían un sindrome genético. Caso clínico: Paciente de 4 años, género femenino, cabello ralo, fascie triangular, fisura palpebral alargada, papadar ojival, mandíbula prominente, escápula alada y clinodactilia del quinto dedo de ambas manos. La prueba molecular de hibridación genómica comparativa por microarreglos, mostró microdeleción de la región 5q35.2 q35.3 de 2.082 MB, que incluye el gen NSD1. Conclusión: Proponemos realizar la prueba de hibridación genómica comparativa en pacientes con retraso global del desarrollo y hallazgos fenotípicos menores.

Sotos Syndrome (SS) is a genetic disease with an autosomal dominant pattern caused by haplo-insufficiency of NSD1 gene secondary to point mutations or microdeletion of the 5q35 locus where the gene is located. It is a rare syndrome, occurring in 7 out of every 100,000 births. The objective of this report is to present the case of a 4 year-old patient with a global developmental delay, as well as specific physical findings suggesting a syndrome of genetic origin. Clinical case: Female patient, 4 years of age, thinning hair, triangular facie, long palpebral fissure, arched palate, prominent jaw, winged scapula and clinodactilia of the fifth finger both hands. The molecular test comparative genomic hybridisation test by microarray was subsequently performed, with the result showing 5q35.2 q35.3 region microdeletion of 2,082 MB, including the NSD1 gene. Conclusion: Finally, this article also proposes the performing of comparative genomic hybridisation as the first diagnostic option in cases where clinical findings are suggestive of SS.

A molecular platform for the diagnosis of multidrug-resistant and pre-extensively drug-resistant tuberculosis based on single nucleotide polymorphism mutations present in Colombian isolates of Mycobacterium tuberculosis

Developing a fast, inexpensive, and specific test that reflects the mutations present in Mycobacterium tuberculosis isolates according to geographic region is the main challenge for drug-resistant tuberculosis (TB) control. The objective of this study was to develop a molecular platform to make a rapid diagnosis of multidrug-resistant (MDR) and extensively drug-resistant TB based on single nucleotide polymorphism (SNP) mutations present in therpoB, katG, inhA,ahpC, and gyrA genes from Colombian M. tuberculosis isolates. The amplification and sequencing of each target gene was performed. Capture oligonucleotides, which were tested before being used with isolates to assess the performance, were designed for wild type and mutated codons, and the platform was standardised based on the reverse hybridisation principle. This method was tested on DNA samples extracted from clinical isolates from 160 Colombian patients who were previously phenotypically and genotypically characterised as having susceptible or MDR M. tuberculosis. For our method, the kappa index of the sequencing results was 0,966, 0,825, 0,766, 0,740, and 0,625 forrpoB, katG, inhA,ahpC, and gyrA, respectively. Sensitivity and specificity were ranked between 90-100% compared with those of phenotypic drug susceptibility testing. Our assay helps to pave the way for implementation locally and for specifically adapted methods that can simultaneously detect drug resistance mutations to first and second-line drugs within a few hours.

Cryptic Insertion of the BCR Gene at 9q34 in Philadelphia-Negative Chronic Myelogenous Leukaemia

Chronic myelogenous leukaemia (CML) is a myeloproliferative neoplasm that is almost always characterised by the presence of t(9;22)(q34;q11.2). Approximately 5% to 10% of CML patients lack cytogenetic evidence of t(9;22)(q34;q11.2) but have the breakpoint cluster region (BCR)/ABL1 fusion, as revealed by fl uorescence in situ hybridisation (FISH) or the reverse transcription-polymerase chain reaction (RT-PCR). We present a case of Philadelphia-negative CML with a cryptic insertion of BCR at 9q34. A 22-year-old woman incidentally presented with marked leucocytosis and anaemia. Her complete blood count results were as follows: white blood cells, 238.61x10(9)/L; haemoglobin, 9.6 g/dL; platelets, 395x10(9)/L. A peripheral blood smear showed leucocytosis with neutrophilia, basophilia, left-shifted neutrophils, and circulating blasts comprising 2% of the total leucocytes. The bone marrow showed a striking increase in megakaryocytes and granulocytic precursors. The myeloid/erythroid ratio was 7.4:1, and blasts comprised up to 1.8% of all nucleated cells. Bone marrow sections revealed active megakaryopoiesis and granulopoiesis with 100% cellularity. Chromosomal analysis revealed a normal karyotype. However, interphase FISH using a dual-colour BCR/ABL1 fusion probe showed an atypical pattern consisting of one red, two green, and one fusion (1R2G1F) signal in 97.5% of the 200 analysed cells. Metaphase FISH revealed a single BCR/ABL1 fusion signal on chromosome 9. RT-PCR was positive for BCR/ABL1 (b3a2). Quantitative PCR revealed a normalised copy number of 15.32. The patient started her treatment with imatinib, reached a complete molecular response eight months afterwards, and has been coping well without any adverse events.

In house reverse membrane hybridisation assay versus GenoType MTBDRplus and their performance to detect mutations in the genes rpoB, katG and inhA

Drug-resistant tuberculosis (TB) threatens global TB control and is a major public health concern in several countries. We therefore developed a multiplex assay (LINE-TB/MDR) that is able to identify the most frequent mutations related to rifampicin (RMP) and isoniazid (INH) resistance. The assay is based on multiplex polymerase chain reaction, membrane hybridisation and colorimetric detection targeting of rpoB and katG genes, as well as the inhA promoter, which are all known to carry specific mutations associated with multidrug-resistant TB (MDR-TB). The assay was validated on a reference panel of 108 M. tuberculosis isolates that were characterised by the proportion method and by DNA sequencing of the targets. When comparing the performance of LINE-TB/MDR with DNA sequencing, the sensitivity, specificity and agreement were 100%, 100% and 100%, respectively, for RMP and 77.6%, 90.6% and 88.9%, respectively, for INH. Using drug sensibility testing as a reference standard, the performance of LINE-TB/MDR regarding sensitivity, specificity and agreement was 100%, 100% and 100% (95%), respectively, for RMP and 77%, 100% and 88.7% (82.2-95.1), respectively, for INH. LINE-TB/MDR was compared with GenoType MTBDRplus for 65 isolates, resulting in an agreement of 93.6% (86.7-97.5) for RIF and 87.4% (84.3-96.2) for INH. LINE-TB/MDR warrants further clinical validation and may be an affordable alternative for MDR-TB diagnosis.

Detección del virus de la tristeza de los cítricos por serología, microscopía e hibridación in situ / In situ serological, microscopy and hybridization detection of CTV

El virus de la tristeza de los cítricos (CTV) es perjudicial para la citricultura y causa la enfermedad llamada tristeza de los cítricos. Infecta las especies del género Citrus ocasionando la muerte de millones de árboles. Los síntomas son decaimiento rápido (QD) y acanalamiento de tallo (SP). En el trabajo se diagnosticó molecular y serológicamente al CTV en aislados provenientes de Citrus aurantifolia o Lima Tahití (LT) y Citrus madurensis (Lour) o Calamondino (Ca), y se realizaron estudios preliminares de detección viral por medio de microscopía óptica e hibridación in situ. Se utilizó IC-RT-PCR e inmunoimpresión de tejido (IMI) expuesto a los anticuerpos monoclonales 3DF1+3CA5, y con el anticuerpo discriminante MCA 13 con técnica de Enzyme Linked Inmunossorbent Assay Doble Sándwich (Elisa-DAS). La detección por microscopía se realizó sobre secciones de pecíolo de LT y C que se tiñeron con Azure A, y con acetato de uranilo y citrato de plomo. Para la hibridación in situ se empleó una sonda marcada con digoxigenina dirigida hacia el gen de la proteína mayor de la cápside. Los resultados de IC-RT-PCR, IMI y Elisa fueron positivos para LT y C, indicando la presencia de variantes virales de tipo severo. Con microscopía de luz se detectaron inclusiones citoplasmáticas en las células acompañantes y del floema, confirmado con IMI y por hibridación in situ. Se visualizaron inclusiones de partículas virales en el tejido vegetal con microscopía electrónica con cambios en la ultraestructura celular como presencia de grandes vacuolas propias de la infección viral. Este trabajo integra distintas técnicas diagnósticas sobre dos especies cítricas exóticas.

Citrus tristeza virus (CTV) is deleterious for citriculture and causes citrus tristeza disease. CTV infects all citrus species thereby causing the death of millions of trees. Its main symptoms are quick decline (QD) and stem pitting (SP). Serological, molecular and microscopy techniques were used in this work for diagnosing CTV in Citrus aurantifolia or Tahiti Lime (Citrus latifolia Tanaka) (TL) and Citrus madurensis (Lour) or Calamondin (Ca) isolates. Petioles were tissue printed (IMI) and exposed to 3DF1+3CA5 monoclonal antibodies; they were then ELISA buffer extracted and exposed to a discriminant MCA 13 monoclonal antibody in a double-antibody sandwich indirect enzyme-linked immunosorbent assay (DASI-ELISA). Immunocapture reverse transcriptase-polymerase chain reaction (IC-RT-PCR) amplification, using specific major coat protein gene (CPG) primers, was used on the ELISA buffer extracts as template. Optical and electron microscopy were used for detection on transversal sections of petiole and stained with azure A, uranyl acetate or lead citrate. Digoxygenin-labelled major CPG CTV probes were used for in situ hybridisation of petioles printing. All IC-RT-PCR, IMI and ELISA results were positive for both LT and C, indicating the presence of severe viral variants. Light microscopy cytoplasm inclusions were detected in the phloem and accompanying cells, confirmed by IMI and in situ hybridisation. Electron microscopy analysis revealed cellular abnormalities with changes in ultrastructure and the presence of big vacuoles which are characteristic of cytoplasmic viral infection. This is the first work integrating all available diagnostic techniques on these two exotic citric species.

Inherited t(9;22) as the cause of DiGeorge syndrome: a case report

DiGeorge syndrome is associated with microdeletion of chromosome 22q11.2. Most cases occur sporadically although vertical transmission has been documented. We report a rare case of DiGeorge syndrome in an 8-year-old girl. Blood sample of the patient was cultured and harvested following standard procedure. All of the 20 cells analysed showed a karyotype of 45,XX,-22,t(9;22)(p23;q11.2). Cytogenetic investigation done on the patient’s mother revealed that she was the carrier for the translocation. Her karyotype was 46,XX,t(9;22)(p23;q11.2). Fluorescence in situ hybridisation (FISH) analysis using TUPLE1 and N25 (Vysis, USA) probes showed deletion of the 22q11.2 region in the patient, confi rming the diagnosis of DiGeorge syndrome. FISH analysis showed no deletion of the region in the mother.

A clinical study of chromosome translocations in extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue in Chinese patients / 中华内科杂志

Objective To investigate the genetic aberrations in extranodal marginal zone lymphoma of mueosa-associated lymphoid tissue (MALT) lymphomas from different sites of the body in Chinese patients. Methods Two hundred and seventeen paraffin-embedded MALT lymphoma specimens from 11 major sites were studied with interphase fluorescence in situ hybridization (FISH) to detect t(11; 18) (q21;q21)/API2-MALT1, t(1; 14) (p22; q32)/IGH-BCL10, (14; 18) (q32; q21)/IGH-MALT1 and BCL6 gene involved chromosome translocations. Results These translocations were mutually exclusive and detected in 21% (46/217) of the cases, including t(11;18) (q21;q21) API2-MALT1 13% (29/217), t (1;14)(p22 ;q32) IGH-BCLIO in 1% (3/217), t(14;18) (q32;q21) IGH-MALT1 1% (2/217), BCL6 involved translocation in 2% (4/217) and IGH-unknown translocation partner in 4% (8/217). t(11; 18) (q21;q21)API2-MALT1 was found with the highest frequency in MALT lymphoma from lungs (47% , 8/17) and small intestine (29%, 4/14), followed by salivary gland (17%, 1/6), stomach (14%, 12/84) and ocular adnexae (6% , 4/68). t(1 ;14) (p22;q32) was only detected in lungs (12%, 2/17) and stomach (1%, 1/84). t(14;18) (q32;q21) was mainly detected in lungs (6%, 1/17) and ocular adnexae (2%, 1/68). BCL6 gene involved translocation was detected in salivary gland (17% , 1/6) and stomach (4%, 3/84). Conclusions It is demonstrated that the four translocatidns occur with markedly variable frequencies in MALT lymphoma of different sites in Chinese patients. The distributions of these chromosome translocations in Chinese patients are slightly different from those reported in western patients.

Identification of Y Chromosomal Material in Turner Syndrome by Fluorescence In Situ Hybridisation (FISH)

Turner syndrome is one of the most common chromosomal abnormalities affecting newborn females. More than half of patients with Turner syndrome have a 45X karyotype. The rest of the patients may have structurally abnormal sex chromosomes or are mosaics with normal or abnormal sex chromosomes. Mosaicism with a second X sex chromosome is not usually of clinical significance. However, Turner syndrome patients having a second Y chromosome or Y chromosomal material are at risk of developing gonadoblastoma later in life. The aim of this study is to compare the results of conventional (karyotyping) and molecular cytogenetics (FISH), and discuss the advantages and limitations in the diagnosis of Turner syndrome. We also aim to compare the degree of mosaicism identified using conventional cytogenetics and FISH techniques. Conventional cytogenetics and FISH analyses were performed on eight peripheral blood samples of patients with Turner syndrome collected between 2004 and 2006. From this study, two out of eight patients with Turner syndrome were found to have the sex determining region on the Y chromosome (SRY) gene by FISH analysis. Our results showed that the rate of detection of mosaic cases in Turner syndrome was also increased to 88% after using the FISH technique. We concluded that FISH is more superior to conventional cytogenetics in the detection of the Y chromosomal material. FISH is also a quick and cost effective method in diagnosing Turner syndrome and assessing the degree of mosaicism.

INCREASED EXPRESSION OF c-myc, c-erbB-2 PROTO-ONCOGENES IN HUMAN THYROID CARCINOMA / 中华内分泌代谢杂志

The expression, amplification and rearrangement of c-myc and c-erbB-2 genes in thyroid tumor were studied. 32P-dATP-labelled probes of c-myc, c-erbB-2 and v-sis DNA fragments were used to hybridise with the cellular total RNA. We found that c-myc and c-erbB-2 oncogenes were expressed in all samples. The levels of c-myc RNA were three-to eleven-fold higher in 9 out of 15 cancer samples when compared with that in normal tissues. In 7 of 15 cancer samples, c-erbB-2 gene was overexpressed 10~50 times higher than normal. No v-sis RNA was detected in all the samples. Southern blot hybridisation showed that rearrangement of c-myc oncogene were observed in 4 cancer samples, of which one showed a 150-fold amplification of c-myc gene. No amplification or rearrangement of c-erbB-2 gene was detected. These data indicate that the activat.:on of c-myc and c-erbB-2 oncogenes may contribute to the development and/or maintenance of the malignant phenotype of the thyroid carcinomas.