Molecular genetics of BCR-ABL1 negative myeloproliferative neoplasms in India

Introduction: Over the past decade, we have moved on from a predominantly morphological and clinical classification of myeloproliferative neoplasms (MPN) to a more evolved classification that accounts for the molecular heterogeneity that is unique to this subgroup of hematological malignancies. This usually incorporates mutations in Janus kinase 2 (JAK2), MPL, and calreticulin (CALR) genes. In this manuscript, we report the frequency of these mutations in a cohort of Indian patients at a tertiary cancer center. Materials and Methods: One hundred and thirty cases of MPN were included in this study. These cases were diagnosed and classified based on the World Health Organization 2008 criteria. JAK2 and MPL mutations were detected using high sensitivity allele-specific polymerase chain reaction using fluorescent labeled primers followed by capillary electrophoresis. A subset of JAK2 and CALR mutations were assessed using a fragment length assay. Results: Among the MPN, we had 20 cases of polycythemia vera (PV), 34 cases of essential thrombocythemia (ET), and 59 of myelofibrosis (MF). JAK2, MPL, and CALR mutations were mutually exclusive of each other. Seventeen cases were categorized as MPN unclassifiable (MPN-U). JAK2p.V617F and MPL mutations were present in 60% (78 of 130) and 5.3% (7 of 130) of all MPN. All the PV cases harbored the JAK2 p.V617F mutation. A total of 23.8% (31 of 130) of patients harbored CALR mutations. CALR exon 9 mutations were detected in 60.8% (14 of 23) and 50% (5 of 10) of JAK2 and MPL negative MF and ET cases, respectively. MPN-U cases included three JAK2 p.V617F positive, two MPL p.W515 L, and 12 CALR positive cases. Ten different types of CALR indels (8 deletions and 2 insertions) were detected of which Type I and Type II mutations were the most common, occurring at a frequency of 45.1% (14 of 31) and 22.5% (7 of 31), respectively. Discussion and Conclusion: We report frequencies of JAK2 p. V617F, MPL exon 10 and CALR mutations in 130 patients similar to those reported in western literature. These mutations carry not only diagnostic but also prognostic relevance.

Patterns and inferred processes associated with sea turtle strandings in Paraíba State, Northeast Brazil / Padrões e inferências associadas com encalhes de tartarugas marinhas no Estado da Paraíba, Nordeste do Brasil

This study analysed sea turtle strandings on the coast of Paraíba State, Northeastern Brazil, from August 2009 to July 2010. A total of 124 strandings were recorded in this period: green turtle Chelonia mydas (n = 106), hawksbill Eretmochelys imbricata (n = 15), olive ridley Lepidochelys olivacea (n = 2) and loggerhead Caretta caretta (n = 1). Of all turtles for which the Curved Carapace Length (CCL) was measured (n = 122), only 12 individuals (9.7%) were adults. Twenty individuals had synthetic anthropogenic debris in the gastrointestinal tract. Other traces of human interactions were observed in 43 individuals, such as injuries caused by entanglement in fishing lines or nets, collisions with vessels, direct contact with oil spills and lesions caused by sharp or spiked objects. Moreover, in 28.5% of the stranded turtles, the presence of external tumors was noticed, suggestive of fibropapillomatosis and in 9.7%, shark bite marks were observed. Of the 107 individuals that were sexed, 76 were females and 31 were males. Most turtles (72.6%) became stranded during the spring/summer (between October and March). We found evidence of human interactions (injuries) in half of the strandings, but in most cases it was not possible to determine if such interactions were the cause of death. A logistic regression found a significant relationship between CCL, ingestion of debris and lesions caused by sharks or spiked objects. Systematic data collection from stranded sea turtles can provide useful biological information, such as seasonal and spatial patterns in their occurrence and mortality, age structure, sex ratio and diet, as well as possible mortality causes.

Este estudo analisou encalhes de tartarugas marinhas na costa do Estado da Paraíba, Nordeste do Brasil, de agosto de 2009 a julho de 2010. Neste período, 124 encalhes foram registrados: tartaruga-verde Chelonia mydas (n = 106), tartaruga-de-pente Eretmochelys imbricata (n = 15), tartaruga-oliva Lepidochelys olivacea (n = 2) e cabeçuda Caretta caretta (n = 1). Dentre todas as tartarugas que tiveram o Comprimento Curvilíneo da Carapaça (CCC) medido (n = 122), apenas 12 indivíduos (9,7%) foram considerados adultos. Vinte indivíduos tinham detritos antropogênicos sintéticos no trato gastrointestinal. Em 43 indivíduos, outros vestígios de interações humanas foram observados, tais como lesões causadas por emaranhamento em linhas ou redes de pesca, colisões com embarcações, contato direto com derramamentos de óleo e lesões causadas por objetos perfurocortantes. Além disso, em 28,5% das tartarugas encalhadas, foi observada a presença de tumores externos sugestivos de fibropapilomatose e em 9,7%, foram observadas marcas de mordidas de tubarão. Dos 107 indivíduos sexados, 76 eram fêmeas e 31 eram machos. A maioria das tartarugas (72,6%) encalhou durante a primavera/verão (entre outubro e março). Foram encontrados sinais de interações humanas (lesões) em metade dos encalhes, mas na maioria dos casos, não foi possível determinar se tais interações foram a causa da morte. A regressão logística encontrou uma relação significativa entre CCC e ingestão de detritos, lesões causadas por objetos perfuro-cortantes e ataques de tubarões. A coleta sistemática de dados de tartarugas marinhas encalhadas pode fornecer informações biológicas úteis, tais como padrões sazonais e espaciais na sua ocorrência e mortalidade, estrutura etária, razão sexual, dieta, bem como possíveis causas de mortalidade.

Ejaculados individuais e pools de sêmen: diferenças em condições experimentais / Individual ejaculates and pooled semen: differences under experimental conditions

Avaliaram-se ejaculados caninos individuais e pools de sêmen submetidos a dois tratamentos de renovação do meio diluidor. Sêmen de seis cães foi coletado, na forma de ejaculados individuais e pools de sêmen, diluído na proporção de 1:1 em meio Tris-Gema, centrifugado a 500g/10min, e o pellet ressuspendido até concentração final de 50x10(6) espermatozoides/mL. O sêmen foi resfriado a 0,26ºC/min, entre 37 e 16ºC, e 0,08ºC/min, entre 16 a 8ºC, e mantido em geladeira a 5ºC por 14 dias. No Tratamento 1, o meio diluidor foi renovado a cada seis dias, e no Tratamento 2 aos 12 dias. O sêmen foi avaliado, a cada 48 horas, quanto à motilidade espermática, utilizando-se o Sperm Class Analyser® (SCA), e quanto à integridade de membranas pelo teste hiposmótico e coloração com PI/CFDA. A formação de pools de sêmen simplificou sua manipulação, principalmente com relação ao aumento do volume da amostra disponível; no entanto, resultados obtidos a partir de ejaculados individuais mostraram diferenças entre tratamentos, não identificadas nos pools de sêmen.

Individual ejaculates and pooled dog semen submitted to two treatments of medium exchange were evaluated. Semen was collected from six dogs, as individual ejaculates and pooled semen, diluted in a 1:1 ratio in Tris-Yolk medium, centrifuged at 500g/10min and ressuspended to the final concentration of 50x10(6) sptz/mL. The samples were cooled at rates of 0.26 o C/min between 37 and 16 o C, and 0.08ºC/min from 16 to 8ºC, and then kept in a refrigerator for 14 days. In Treatement 1 medium was exchanged every six days and in Treatment 2 after twelve days. The cooled samples were evaluated every 48 hours for sperm motility using the Sperm Calss Analyser® (SCA), and membrane integrity with hiposmotic swelling test and PI/CFDA stain. Pooled semen was easier to handle, mainly considering the decreased work due to volume. When submitted to medium exchange, pooled semen behaved similarly to individual ejaculates; however, results obtained from individual ejaculates showed differences between treatments, which were not apparent in pooled semen results.