RESUMO
Oftentimes ignored or infrequently expressed, some transgender persons harbor a desire for parenthood. Given the evolution of medical techniques and the enacting of legislative reforms, it is henceforth possible to propose fertility preservation strategies in the overall context of gender transidentity. During the "female to male" (FtM) transition pathway, androgen therapy has an impact on gonadic function, generally inducing blockage of the ovarian function, with amenorrhea. Even though these events may be reversed on cessation of treatment, the possible long-term effects on future fertility and on the health of children yet to be born are little known. Moreover, transition surgeries definitively compromise the possibility of pregnancy insofar as they involve bilateral adnexectomy and/or hysterectomy. Options for fertility preservation in the framework of FtM transition are premised on cryopreservation of oocytes and/or ovarian tissue. In a comparable manner, even though relevant documentation is lacking, hormonal treatments for persons transitioning from male to female (MtF) can have an impact on future fertility. In the event of surgery involving bilateral orchidectomy in which spermatozoid cryopreservation has not been carried out, fertility is definitively impossible. In both cases and under present-day legislation, numerous legal and regulatory barriers render highly problematic the reutilization of cryopreserved gametes. Given these different constraints, it is indispensable to closely supervise these types of treatment by proposing psychological support.
Assuntos
Preservação da Fertilidade , Pessoas Transgênero , Criança , Humanos , Masculino , Feminino , Preservação da Fertilidade/métodos , Preservação da Fertilidade/psicologia , Criopreservação/métodos , Pessoas Transgênero/psicologiaRESUMO
OBJECTIVES: The aim of this study is to understand the changes within families during confinement motivated by the COVID-19 pandemic and to explore the psycho-emotional experiences of children and their parents in this new situation. Confinement necessarily induced significant changes in daily family routines, particularly for work, education, leisure and social activities. In the more vulnerable pediatric population, several authors have warned of the need to consider the impact of lockdown measures during COVID-19 on the psychological impact and well-being. METHOD: This is an anonymous online survey with methodology combining quantitative and qualitative analyses. The questions targeted several themes such as life context, emotional experience and the impact on daily habits in children and adolescents, as perceived by parents. Participants are adults and parents of at least one child. They were recruited through social media and email. RESULTS: A total of 439 parents responded to the questionnaire. The families generally stayed in their usual place of residence and managed to adapt well. On average, the children's level of worry (as estimated by parents) was lower than the level of worry parents attributed to themselves. For the majority, the parents did not observe any change, the psychological state of the children and adolescents was generally stable, but for those who experienced more negative emotions than usual, it was an increase in boredom, irritability and anger. A decrease in the quality of sleep was also observed by a third of the respondents. On the other hand, an increase in autonomy was noted. Regarding the quality of family cohabitation, an important result showed that confinement had improved family relationships for 41% parents but at the expense of usual social ties inducing a feeling of deprivation. Indeed, the participants evoke a lack of "social link" and "social contact with friends". Lack became synonymous with absence, a feeling of loneliness and separation. CONCLUSION: Our results confirm European and international data collected in children in countries where strict lockdown measures have been applied. Despite the negative emotions felt in some children, confinement has helped develop new resources in most families. Families seem to have been successful in maintaining a stable and secure routine which has certainly been a protective factor against anxiety. Some reported factors, such as bonding, could be protective factors and constitute good leads in interventions to be offered to children and their families.
Assuntos
COVID-19 , Adolescente , Adulto , Criança , Humanos , Pandemias , Controle de Doenças Transmissíveis , Pais/psicologia , FamíliaRESUMO
Medical device made to measure by 3D printing are now emerging in hospital. In order to improve the precision of surgery and facilitate the treatment of complicated cases, patient specific surgical guides for dental implantology are made by stereolithography in our facial surgical unit. This new activity requires to ensure the safety of patients and health personnel by validating the various step of the manufacturing circuit. In this context, the goal of this work was to study the quality of autoclave sterilisation of the patient specific surgical guide made to measure in our hospital. A protocol of sterility test was designed and validated. Sterility of implantology guides 0, 7, 14 and 28 days after sterilisation was checked. The impact of the autoclave sterilisation on the medical device structure was evaluated by visual check and during surgeries. The sterility of the implantology guides up to 28 days after sterilisation was also validated. The protocol of sterility test executed can be extended to other hospitals interested in validating a sterility test. No deformation was observed by surgeons during the dental implant process. Future studies may be necessary to check the accurate impact of sterilisation on surgical guide structure.
Assuntos
Infertilidade , Cirurgia Assistida por Computador , Desenho Assistido por Computador , Hospitais , Humanos , EsterilizaçãoRESUMO
AIMS: This study aimed to evaluate in vitro efficacy of essential oils (EOs) and their compounds (EOCs) alone or in combination against Aeromonas salmonicida subsp. salmonicida, the causative agent of furunculosis in salmonid fish. METHODS AND RESULTS: Antimicrobial activity of 13 EOs and 16 EOCs was investigated for four A. salmonicida subsp. salmonicida strains using broth microdilution. The checkerboard assay was used to evaluate a putative synergy between the most efficient EOs and EOCs against the tested strains. Cinnamon bark, oregano, clove, and thyme oils and their major compounds cinnamaldehyde, eugenol, carvacrol and thymol showed the lower minimum inhibitory concentration and minimum bactericidal concentration values. The association of cinnamaldehyde and eugenol (V/V: 30%/70%) showed a synergistic activity against three tested strains. The combinations of cinnamon with oregano, clove or thyme EOs showed a neutral or additive activity against all the tested strains. CONCLUSIONS: Cinnamon, oregano, clove and thyme oils and their major phytochemical compounds showed strong activities against A. salmonicida subsp. salmonicida strains. SIGNIFICANCE AND IMPACT OF THE STUDY: To reduce the use of antibiotics in aquaculture, phytochemicals such as cinnamaldehyde and eugenol can be tested alone or in combination in in vivo studies as functional feed alternatives.
Assuntos
Aeromonas salmonicida/efeitos dos fármacos , Antibacterianos/farmacologia , Óleos Voláteis/farmacologia , Compostos Fitoquímicos/farmacologia , Animais , Sinergismo Farmacológico , Furunculose/microbiologia , Testes de Sensibilidade Microbiana , Óleos Voláteis/química , Óleos de Plantas/química , Óleos de Plantas/farmacologia , Salmonidae/microbiologiaRESUMO
The goal of this retrospective study is to assess if diabetic patients are more likely to develop urinary tract infection (UTI) within the context of ureteral obstruction. 804 patients that had received an emergency treatment by double J placement for ureteral stone were selected between January 2004 and December 2014 at the Clinique Saint Pierre d'Ottignies, Ottignies-Louvain-La-Neuve,Belgium. They were divided in two groups : patients with UTI associated and the control group with the non infected ones. In the group of infected patients, 82 were diabetic whereas they were 46 in the control group. There was a significant difference regarding the presence of diabetes between the group of patients with UTI and the control group (p inferior to 0.001). This study demonstrates that diabetic patients are at higher risk of urinary tract infection in case of ureteral obstruction, thus an invasive treatment could be considered faster.
Cette étude rétrospective a pour but de déterminer si les patients diabétiques sont plus à risque de développer une infection urinaire en cas d'obstruction urétérale. 804 patients ayant bénéficié de la mise en place d'une sonde JJ en urgence pour une lithiase urétérale ont été sélectionnés entre le 1er janvier 2004 et le 31 décembre 2014 à la Clinique Saint Pierre d'Ottignies, à Ottignies Louvain la Neuve, Belgique. Ces patients ont été répartis en 2 groupes suivant qu'ils étaient ou non suspects d'infection urinaire associée. Dans le groupe des patients infectés, 82 patients étaient diabétiques alors qu'ils étaient 46 dans le groupe des patients sans infection urinaire. Les résultats montrent qu'il existe une différence significative entre les deux groupes en présence de diabète (p inf�rieur a 0,001). Cette étude montre un risque accru d'infection urinaire en cas d'obstruction urétérale chez les patients diabétiques. Un traitement invasif pourrait donc être envisagé plus rapidement.
Assuntos
Complicações do Diabetes , Diabetes Mellitus , Obstrução Ureteral , Infecções Urinárias , Bélgica , Humanos , Estudos Retrospectivos , Fatores de Risco , Obstrução Ureteral/complicações , Infecções Urinárias/etiologiaRESUMO
It is generally believed that neural damage that occurs early in development is associated with greater adaptive capacity relative to similar damage in an older individual. However, few studies have surveyed whole brain changes following early focal damage. In this report, we employed multimodal magnetic resonance imaging analyses of adult rhesus macaque monkeys who had previously undergone bilateral, neurotoxic lesions of the amygdala at about 2 weeks of age. A deformation-based morphometric approach demonstrated reduction of the volumes of the anterior temporal lobe, anterior commissure, basal ganglia, and pulvinar in animals with early amygdala lesions compared to controls. In contrast, animals with early amygdala lesions had an enlarged cingulate cortex, medial superior frontal gyrus, and medial parietal cortex. Diffusion-weighted imaging tractography and network analysis were also used to compare connectivity patterns and higher-level measures of communication across the brain. Using the communicability metric, which integrates direct and indirect paths between regions, lesioned animals showed extensive degradation of network integrity in the temporal and orbitofrontal cortices. This work demonstrates both degenerative as well as progressive large-scale neural changes following long-term recovery from neonatal focal brain damage.
Assuntos
Tonsila do Cerebelo/patologia , Lesões Encefálicas/fisiopatologia , Vias Neurais/fisiologia , Tonsila do Cerebelo/diagnóstico por imagem , Animais , Animais Recém-Nascidos , Lesões Encefálicas/induzido quimicamente , Lesões Encefálicas/diagnóstico por imagem , Lesões Encefálicas/patologia , Conectoma , Imagem de Difusão por Ressonância Magnética , Agonistas de Aminoácidos Excitatórios/toxicidade , Feminino , Ácido Ibotênico/toxicidade , Processamento de Imagem Assistida por Computador , Estudos Longitudinais , Macaca mulatta , MasculinoRESUMO
Babesiosis is an important veterinary and zoonotic tick borne disease caused by the hemoprotozoan Babesia spp. which infects red blood cell of its vertebrate host. In order to control the infection, vaccination that targets molecules involved in the invasion process of red blood cells could provide a good alternative to chemotherapy. Among these molecules, Apical Membrane Antigen-1 (AMA-1) has been described as an excellent vaccine candidate in Plasmodium spp. In this paper, we have investigated AMA-1 of Babesia divergens (BdAMA-1) as vaccine candidate by evaluating its polymorphism and by studying the humoral response against BdAMA-1 of sheep experimentally infected with B. divergens. Polymorphism of BdAMA-1 was investigated by sequencing the corresponding gene of 9 B. divergens isolates from different geographical areas in France. Two Bdama-1 haplotypes (A and B) could be defined based on 2 non-synonymous point mutations. In silico prediction of linear epitopes revealed that the antigenicity of the 2 haplotypes is very similar. Antibody production against the extracellular domain of BdAMA-1 is weak and late, between 1 and 5 months after the inoculation of parasites. Both IgG1 and IgG2 are components of the anti-BdAMA-1 response. These results indicate that while BdAMA-1 may not be an immuno-dominant antigen, it could induce a mixed type 1 and type 2 immune response. In light of these results, the potential of BdAMA-1 as vaccine candidate is discussed.
Assuntos
Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/imunologia , Babesia/imunologia , Babesiose/imunologia , Proteínas de Protozoários/imunologia , Vacinas Protozoárias , Animais , Anticorpos Antiprotozoários/biossíntese , Babesia/genética , Babesiose/parasitologia , Babesiose/prevenção & controle , Bovinos , DNA de Protozoário/química , DNA de Protozoário/isolamento & purificação , Humanos , Reação em Cadeia da Polimerase , Polimorfismo Genético , Vacinas Protozoárias/imunologia , Vacinas Protozoárias/normas , Proteínas Recombinantes/imunologia , Alinhamento de Sequência , OvinosRESUMO
DNA microarrays were rapidly scaled up from 256 to 6.5 million targets, and although antibody microarrays were proposed earlier, sensitive multiplex sandwich assays have only been scaled up to a few tens of targets. Cross-reactivity, arising because detection antibodies are mixed, is a known weakness of multiplex sandwich assays that is mitigated by lengthy optimization. Here, we introduce (1) vulnerability as a metric for assays. The vulnerability of multiplex sandwich assays to cross-reactivity increases quadratically with the number of targets, and together with experimental results, substantiates that scaling up of multiplex sandwich assays is unfeasible. We propose (2) a novel concept for multiplexing without mixing named antibody colocalization microarray (ACM). In ACMs, both capture and detection antibodies are physically colocalized by spotting to the same two-dimensional coordinate. Following spotting of the capture antibodies, the chip is removed from the arrayer, incubated with the sample, placed back onto the arrayer and then spotted with the detection antibodies. ACMs with up to 50 targets were produced, along with a binding curve for each protein. The ACM was validated by comparing it to ELISA and to a small-scale, conventional multiplex sandwich assay (MSA). Using ACMs, proteins in the serum of breast cancer patients and healthy controls were quantified, and six candidate biomarkers identified. Our results indicate that ACMs are sensitive, robust, and scalable.
Assuntos
Anticorpos/análise , Biomarcadores Tumorais/sangue , Neoplasias da Mama/sangue , Proteínas de Neoplasias/sangue , Análise Serial de Proteínas/métodos , Adulto , Idoso , Proteínas Sanguíneas/análise , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Pessoa de Meia-Idade , Reprodutibilidade dos TestesRESUMO
Scholars have argued for centuries that affective states involve interoception, or representations of the state of the body. Yet, we lack a mechanistic understanding of how signals from the body are transduced, transmitted, compressed, and integrated by the brains of humans to produce affective states. We suggest that to understand how the body contributes to affect, we first need to understand information flow through the nervous system's interoceptive pathways. We outline such a model and discuss how unique anatomical and physiological aspects of interoceptive pathways may give rise to the qualities of affective experiences in general and valence and arousal in particular. We conclude by considering implications and future directions for research on interoception, affect, emotions, and human mental experiences.
Assuntos
Encéfalo , Interocepção , Humanos , Interocepção/fisiologia , Encéfalo/fisiologia , Afeto/fisiologia , Neurobiologia , Emoções/fisiologiaRESUMO
The evolution of medical care for transgender people currently makes it possible to propose fertility preservation. Fertility preservation should be initiated before the start of hormonal treatments and/or surgical procedures. The "reproductive" aspect and the desire for parenthood among transgender people have long been ignored. However, these aspects are important to consider and fertility preservation should be discussed before the start of the physical transition. The aim of this review is to assess the literature on fertility preservation for transgender women ("male to female"). Many uncertainties remain regarding the impact of hormonal treatments on the reproductive functions of transgender women and their reversibility. However, the significant increase in the number of recently published articles is evidence of the improvement in the conditions of access to these procedures for women starting a transition process. Nevertheless, there are still a number of barriers that can prevent or delay this process.
Assuntos
Preservação da Fertilidade , Pessoas Transgênero , Criopreservação , Feminino , Humanos , Masculino , Exame FísicoRESUMO
Ovine babesiosis is an important disease in China and responsible for economic losses. Several Babesia strains are involved, but Babesia sp. BQ1 (Lintan) and Babesia sp. BQ1 (Ningxian) are particularly prevalent in the Guansu region. Babesia divergens, in contrast, can experimentally infect spleen-intact sheep, but does not induce clinical signs. The immune response of spleen-intact sheep to Babesia sp. BQ1 (Lintan) and to B. divergens was therefore compared to identify the immune mechanisms involved in pathogenicity. The greater pathogenicity of Babesia sp. BQ1 (Lintan) than that of B. divergens was confirmed: sheep infected with Babesia sp. BQ1 (Lintan), but not with B. divergens, developed hyperthermia and showed patent parasitaemia in Giemsa-stained blood smears from the ear vein. Furthermore, more parasites were also detected in the blood from the jugular vein of Babesia sp. BQ1 (Lintan)-infected sheep. Pathogenicity of Babesia spp. involved cellular responses, but not humoral responses. Interferon-gamma was produced only by specifically activated PBMC from B. divergens-infected sheep and interleukin-10 only by specifically activated PBMC from Babesia sp. BQ1 (Lintan)-infected sheep. The role of these cytokines in the course of infection by Babesia spp. is discussed.
Assuntos
Babesia/patogenicidade , Babesiose/veterinária , Interferon gama/imunologia , Interleucina-10/imunologia , Doenças dos Ovinos/patologia , Doenças dos Ovinos/parasitologia , Animais , Babesia/imunologia , Babesiose/imunologia , Babesiose/patologia , Células Cultivadas , China , Febre , Humanos , Interferon gama/toxicidade , Interleucina-10/toxicidade , Leucócitos Mononucleares/imunologia , Parasitemia , Doenças dos Ovinos/imunologiaRESUMO
Babesia sp. BQ1 (Lintan) is a Babesia isolated from sheep infested with Haemaphysalis qinghaiensis in China, and is closely related to B. motasi based on the 18S rRNA gene sequence. In the present study, an ELISA was developed with merozoite antigens of Babesia sp. BQ1 (Lintan) (BQMA) purified from in vitro culture. When the positive threshold was chosen as 30% of the antibodies rate, evaluated with 198 negative sera, the specificity was 95.5%. Except for Babesia sp. Tianzhu, there was no cross-reaction between BQMA and positive sera from Babesia sp. BQ1 (Ningxian)-, Babesia sp. Hebei-, Babesia sp. Xinjiang-, Theileria luwenshuni-, T. uilenbergi-, or Anaplasma ovis-infected sheep, which are the dominant haemoparasites of small ruminants in China. Specific antibodies against Babesia sp. BQ1 (Lintan) were produced 1 or 2 weeks post-infection and a high level of antibodies persisted for more than 8 months in experimentally infected sheep. This ELISA was tested on 974 sera collected from field-grazing sheep in 3 counties of Gansu province, northwestern China to evaluate the seroprevalence of Babesia sp. BQ1 (Lintan) infection and the average positive rate was 66.84%. The feasibility of increasing the specificity of this BQMA-based ELISA, by using some BQMA antigens for serodiagnosis is discussed.
Assuntos
Babesia/metabolismo , Merozoítos/metabolismo , Proteínas de Protozoários/metabolismo , Animais , Anticorpos Antiprotozoários/sangue , Babesia/genética , Babesiose/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Regulação da Expressão Gênica/fisiologia , Proteínas de Protozoários/genética , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Testes Sorológicos/veterinária , Ovinos , Doenças dos Ovinos/sangue , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/parasitologiaRESUMO
The digital revolution has led to many recent developments in implantology that have considerably facilitated implant planning and the creation of surgical guides. The purpose of this article is to explain how we set up a digital workflow in a large city hospital and how we met the requirements of the European regulations on the production of custom-made devices in a medical establishment. The internal manufacture of a surgical guide complied with European regulation EU/2017/45 concerning medical devices. This regulation allowed the hospital to create these medical devices locally without CE marking. However, the hospital must be declared as a manufacturer of medical devices and comply with the general requirements in terms of safety and performance related to the manufacture and use of medical devices. In addition, hospitals are large structures involving many different actors. Each step of the digital workflow, which included both the patient course and the creation of the surgical guide, was thus adapted to European regulations by considering local constraints.
Assuntos
Implantação Dentária Endóssea , Cirurgia Assistida por Computador , Desenho Assistido por Computador , HumanosRESUMO
Polymerase chain reaction (PCR) assessment of clonal T-cell receptor (TCR) and immunoglobulin (Ig) gene rearrangements is an important diagnostic tool in mature T-cell neoplasms. However, lack of standardized primers and PCR protocols has hampered comparability of data in previous clonality studies. To obtain reference values for Ig/TCR rearrangement patterns, 19 European laboratories investigated 188 T-cell malignancies belonging to five World Health Organization-defined entities. The TCR/Ig spectrum of each sample was analyzed in duplicate in two different laboratories using the standardized BIOMED-2 PCR multiplex tubes accompanied by international pathology panel review. TCR clonality was detected in 99% (143/145) of all definite cases of T-cell prolymphocytic leukemia, T-cell large granular lymphocytic leukemia, peripheral T-cell lymphoma (unspecified) and angioimmunoblastic T-cell lymphoma (AILT), whereas nine of 43 anaplastic large cell lymphomas did not show clonal TCR rearrangements. Combined use of TCRB and TCRG genes revealed two or more clonal signals in 95% of all TCR clonal cases. Ig clonality was mostly restricted to AILT. Our study indicates that the BIOMED-2 multiplex PCR tubes provide a powerful strategy for clonality assessment in T-cell malignancies assisting the firm diagnosis of T-cell neoplasms. The detected TCR gene rearrangements can also be used as PCR targets for monitoring of minimal residual disease.
Assuntos
Genes de Imunoglobulinas , Leucemia de Células T/genética , Linfoma de Células T/genética , Reação em Cadeia da Polimerase/métodos , Receptores de Antígenos de Linfócitos T/genética , Amplificação de Genes , Rearranjo Gênico , Genótipo , Humanos , Imuno-Histoquímica , Leucemia Prolinfocítica/genética , Leucemia Prolinfocítica/imunologia , Leucemia Prolinfocítica/patologia , Leucemia de Células T/imunologia , Leucemia de Células T/patologia , Linfoma Difuso de Grandes Células B/genética , Linfoma Difuso de Grandes Células B/imunologia , Linfoma Difuso de Grandes Células B/patologia , Linfoma de Células T/imunologia , Linfoma de Células T/patologia , Linfócitos T/imunologiaAssuntos
Lipopolissacarídeos , Tuberculose , Adulto , Humanos , Lipopolissacarídeos/urina , Tuberculose/urinaRESUMO
In this paper, a complete analytical resolution of the one dimensional Burgers equation with the elastic forcing term -k2x + f(t), k is an element of R is presented. Two methods existing for the case K=0 are adapted and generalized using variable and functional transformations, valid for all values of space and time. The emergence of a Fokker-Planck equation in the method allows the establishment of a connection between the Burgers equation and the Ornstein-Uhlenbeck process.
RESUMO
Humoral and cellular immune responses to Fasciola gigantica experimental infection in buffaloes were studied. The results showed that 33.4+/-9.1% of the infection dose was recovered as adult flukes from infected animals at necropsy. Significant differences of weight gain between infected and non-infected buffaloes was observed at 4 MPI (months post-infection). Anti F. gigantica excretory-secretory products (FgESP)-IgG levels increased significantly from 3 WPI (weeks post-infection) and displayed a peak at 13 WPI. Western blot indicated that in FgESP six major bands of 11.5, 19.0, 23.4, 29.8, 47.5 and 53.2kDa were recognized by F. gigantica-infected buffaloes sera after 0 WPI. Eosinophil numbers increased significantly from 3 WPI in F. gigantica-infected buffaloes and displayed a peak at 8 WPI. Peripheral blood mononuclear cells (PBMC) proliferation induced by FgESP increased from 2 WPI with a peak at 5 WPI. IFNgamma secretion by FgESP-stimulated PBMC appeared early from 1 WPI with three peaks at 2, 5 and 8 WPI, respectively. IL-10 production was observed from 2 WPI with two peaks at 4 and 9 WPI, respectively. Our results suggested that buffaloes were highly susceptible to F. gigantica infection, and this susceptibility could be associated with the late and weak cellular immune response in the early phase of infection and the Th0-like response throughout the infection.
Assuntos
Búfalos/imunologia , Búfalos/parasitologia , Fasciola/imunologia , Fasciolíase/imunologia , Fasciolíase/veterinária , Animais , Anticorpos Anti-Helmínticos/sangue , Formação de Anticorpos/imunologia , Antígenos de Helmintos/imunologia , Western Blotting/veterinária , Búfalos/sangue , Proliferação de Células , Ensaio de Imunoadsorção Enzimática/veterinária , Fasciolíase/sangue , Fasciolíase/parasitologia , Imunidade Celular/imunologia , Interferon gama/imunologia , Interleucina-10/imunologia , Cinética , Contagem de Leucócitos/veterinária , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/imunologia , Fígado/parasitologia , Estatísticas não Paramétricas , Aumento de PesoRESUMO
INTRODUCTION: Standardization of BCR-ABL1 messenger RNA quantification by real-time PCR on the International Scale (IS) is critical for monitoring therapy response in chronic myelogenous leukaemia. Since 2006, BCR-ABL1 IS standardization is propagated along reference laboratories by calculating a laboratory-specific conversion factor (CF), co-ordinated in Europe through the European Treatment and Outcome Study project. Although this process has proven successful to some extent, it has not been achievable for all laboratories due to the complexity of the process and the stringent requirements in terms of numbers of samples to be exchanged. In addition, several BCR-ABL1 IS quantification methods and secondary reference materials became commercially available. However, it was observed that different IS methods generate consistently different results. METHODS: To overcome these difficulties, we have developed an alternative and simple approach of CF calculation, based on the retrospective analysis of existing external quality assessment (EQA) data. Our approach does not depend on the exchange of samples and is solely based on the mathematical CF calculation using EQA results. RESULTS AND CONCLUSION: We have demonstrated by thorough statistical validation that this approach performs well in converting BCR-ABL1 measurements to improve IS estimation. In expectation of a true golden standard method for BCR-ABL1 IS quantification, the proposed method is a valuable alternative.
Assuntos
Proteínas de Fusão bcr-abl/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , RNA Mensageiro/análise , Testes Genéticos , Cooperação Internacional , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Métodos , Variações Dependentes do Observador , Padrões de Referência , Estudos RetrospectivosRESUMO
Southern blot analysis of T cell receptor (TCR) gene rearrangements has proven to be a helpful tool to establish clonality in T cell leukemias and lymphomas. To improve the detection of clonal TCR gamma (TCRG) gene rearrangements by Southern blot analysis, we designed four new Jgamma probes and determined the most optimal restriction enzymes to be used with these probes. Based on detailed analysis of the sequences as well as on hybridization experiments with the TCRGJ21 probe, the Jgamma1.2 and Jgamma2.1 downstream areas were found to be highly homologous, suggesting that during evolution the duplication of the Jgamma region was followed by deletion of the tentative Jgamma2.2 gene segment. Southern blot analysis of 51 T cell acute lymphoblastic leukemias (T-ALL) revealed that all TCRG gene rearrangements can be detected by use of the TCRGJ13 probe in EcoRI digests and the TCRGJ21 probe in PstI digests. Additional probes and digests allow a more precise identification of the exact type of TCRG gene rearrangements in the majority of cases. Almost 90% of the TCRG gene rearrangements in T-ALL involved the Jgamma2 region (16% Jgamma2.1 and 72% Jgamma2.3), whereas Jgamma1 region rearrangements were particularly found in TCRgammadelta+ T-ALL. This information has implications for design of primer sets for PCR analysis at diagnosis and for PCR target choice in detection of minimal residual disease during follow-up of T-ALL patients.