Mixed-phenotype acute leukemia characteristics: first report from Iran

Mixed-phenotype acute leukemia (MPAL) is the infrequent type of acute leukemia characterized by immunophenotypic and/or cytochemical features of both lineages, but the diagnosis of this disease still is a challenge. In this study, we analyzed immunophenotyping, cytochemistry and frequency of MPAL patients to better diagnosis of MPAL characteristics according to WHO 2016 criteria for the first time in Iran. In this retrospective study, 27 patients were diagnosed as MPAL based on WHO 2016 criteria during 2014�2017. Flow cytometric immunophenotyping was performed on PB and BM samples evaluation of different CD marker expressions in MPAL subsets. RT-PCR was performed for the analyses of BCR/ABL1 fusion in MPAL subsets. Among 27 cases, (70.4) 19 cases were B + My, (22.22) 6 cases were T + My, and 2 cases (7.40) were B + T + My. CD34, CD19, HLA-DR, TdT, CD22, iMPO were positive in majority of B + My cases. CD45, iMPO, iCD3, CD7, CD2 and CD5 were positive in majority of T + My cases. HLA-DR, TdT, CD10, CD22, iCD79a, iMPO, CD45, iCD3, CD7, CD3, CD2, CD5 were positive in majority of B + T + My cases. BCR/ABL1 fusion was positive for 3 cases (11.1) of p190 fusion and 2 cases (7.4) of p210 fusion in B + My cases. WHO 2016 criteria are the current standard for diagnosing MPAL. Also, evaluation of TdT, CD2, CD5, CD7 expressions by flow cytometry in EGIL criteria is useful for the better diagnosis of MPAL subsets. In addition, evaluation of BCR/ABL1 and MLL rearrangements in patients should be part of standard work-up in MPAL. © 2018 Springer Nature Switzerland A

Cerebellar Microstructural Abnormalities in Parkinson’s Disease: a Systematic Review of Diffusion Tensor Imaging Studies

Diffusion tensor imaging (DTI) is now having a strong momentum in research to evaluate the neural fibers of the CNS. This technique can study white matter (WM) microstructure in neurodegenerative disorders, including Parkinson’s disease (PD). Previous neuroimaging studies have suggested cerebellar involvement in the pathogenesis of PD, and these cerebellum alterations can correlate with PD symptoms and stages. Using the PRISMA 2020 framework, PubMed and EMBASE were searched to retrieve relevant articles. Our search revealed 472 articles. After screening titles and abstracts, and full-text review, and implementing the inclusion criteria, 68 papers were selected for synthesis. Reviewing the selected studies revealed that the patterns of reduction in cerebellum WM integrity, assessed by fractional anisotropy, mean diffusivity, radial diffusivity, and axial diffusivity measures can differ symptoms and stages of PD. Cerebellar diffusion tensor imaging (DTI) changes in PD patients with “postural instability and gait difficulty” are significantly different from “tremor dominant” PD patients. Freezing of the gate is strongly related to cerebellar involvement depicted by DTI. The “reduced cognition,” “visual disturbances,” “sleep disorders,” “depression,” and “olfactory dysfunction” are not related to cerebellum microstructural changes on DTI, while “impulsive-compulsive behavior” can be linked to cerebellar WM alteration. Finally, higher PD stages and longer disease duration are associated with cerebellum white matter alteration depicted by DTI. Depiction of cerebellar white matter involvement in PD is feasible by DTI. There is an association with disease duration and severity and several clinical presentations with DTI findings. This clinical-imaging association may eventually improve disease management

Bright light therapy and early morning attention, mathematical performance, electroencephalography and brain connectivity in adolescents with morning sleepiness.

Adolescents typically sleep too little and feel drowsy during morning classes. We assessed whether morning use of an LED bright light device could increase alertness in school students. Twenty-six (8M/18F) healthy, unmedicated participants, ages 13-18 years, (mean 17.1±1.4) were recruited following screenings to exclude psychopathology. Baseline assessments were made of actigraph-assessed sleep, attention, math solving ability, electroencephalography and structural and functional MRI (N = 10-11, pre-post). Participants nonrandomly received 3-4 weeks of bright light therapy (BLT) for 30 minutes each morning and used blue light blocking glasses for 2 hours before bedtime. BLT devices were modified to surreptitiously record degree of use so that the hypothesis tested was whether there was a significant relationship between degree of use and outcome. They were used 57±18% (range 23%-90%) of recommended time. There was a significant association between degree of use and: (1) increased beta spectral power in frontal EEG leads (primary measure); (2) greater post-test improvement in math performance and reduction in errors of omission on attention test; (3) reduced day-to-day variability in bed times, sleep onset, and sleep duration during school days; (4) increased dentate gyrus volume and (5) enhanced frontal connectivity with temporal, occipital and cerebellar regions during Go/No-Go task performance. BLT was associated with improvement in sleep cycle consistency, arousal, attention and functional connectivity, but not sleep onset or duration (primary measures). Although this was an open study, it suggests that use of bright morning light and blue light blocking glasses before bed may benefit adolescents experiencing daytime sleepiness. Clinical trial registration: Clinicaltrials.gov ID-NCT05383690

Estimating Young's modulus of zona pellucida by micropipette aspiration in combination with theoretical models of ovum

The zona pellucida (ZP) is the spherical layer that surrounds the mammalian oocyte. The physical hardness of this layer plays a crucial role in fertilization and is largely unknown because of the lack of appropriate measuring and modelling methods. The aim of this study is to measure the biomechanical properties of the ZP of human/mouse ovum and to test the hypothesis that Young's modulus of the ZP varies with fertilization. Young's moduli of ZP are determined before and after fertilization by using the micropipette aspiration technique, coupled with theoretical models of the oocyte as an elastic incompressible half-space (half-space model), an elastic compressible bilayer (layered model) or an elastic compressible shell (shell model). Comparison of the models shows that incorporation of the layered geometry of the ovum and the compressibility of the ZP in the layered and shell models may provide a means of more accurately characterizing ZP elasticity. Evaluation of results shows that although the results of the models are different, all confirm that the hardening of ZP will increase following fertilization. As can be seen, different choices of models and experimental parameters can affect the interpretation of experimental data and lead to differing mechanical properties

in Ph+BCR-ABL1 acute lymphoblastic leukemia the e13a2 (B2A2) transcript is prevalent

Philadelphia-chromosome positive (Ph+), BCR-ABL1+ acute lymphoblastic leukemia (ALL) is a distinct entity that is characterized by specific genomic alterations, low sensitivity to chemotherapy, unstable responsiveness to tyrosine kinase inhibitors (TKIs), and a poor prognosis [1–3]. The frequency of Ph+ ALL varies with age, ranging from <10% in children to about 50% in the elderly [3, 4]. Ph+ ALL is driven by a reciprocal translocation between chromosome 9 and chromosome 22, leading to the formation of hybrid fusion genes, that are all leukemogenic but can vary depending on the site of the breakpoints [5, 6]. The most common gene, that accounts for about 70–80% of all cases, results from the fusion of BCR exon 1 on chromosome 22 with ABL1 exon 2 on chromosome 9, much more rarely with ABL1 exon 3. The resulting e1a2 (or e1a3) fusion gene codes for a leukemogenic protein of 185–190 kd (P190). In 20–30% of cases, the breakpoint on chromosome 22 is downstream to BCR exon 1, resulting from the fusion of ABL1 exon 2 on chromosome 9 with either BCR exon 13 or BCR exon 14 on chromosome 22. The resulting fusion transcripts are named e13a2 (also known as b2a2), and e14a2 (also known as B3A2), respectively. The fusion e13a2 can give rise to only one transcript (e13a2) and one leukemogenic protein of 210 kd (P210). The fusion e14a2 usually give rise to one transcript (e14a2) that is longer, and codes for a leukemogenic protein that is also called P210, but has 25 additional amino acids and different secondary structure elements [5, 6]. Sometimes the e14a2 fusion gene, that retains BCR exon 13, can also generate a minor amount of e13a2 transcript, because of alternative splicing mechanisms. Therefore, the BCR-ABL1P210+ leukemias have two different molecular signatures:e13a2, and e14a2 (+_e13a2). The frequency of the two signatures has never been systematically investigated. In a paper from the MD Anderson Cancer Center it was reported that of 17 patients with Ph+, BCR-ABL1P210+ 76% expressed e13a2 [7]. In a previous international study of the frequency of the two transcripts in 45,503 patients with newly diagnosed BCR-ABL1P210+ chronic myeloid leukemia (CML), it was found that 17,216 patients (37.9%) expressed only e13a2, with a proportion that varied with age, from 39.6% in children and adolescents down to 31.6% in the elderly, and with sex, from 36.5%in females up to 39.2%inmales [8]. To assess the frequency of e13a2 in BCR-ABL1P210+ ALL, we collected the molecular data from 849 patients with Ph+, BCR-ABL1P210+ ALL, from 39 Institutions, as a first quick step of a project that was designed to evaluate the clinical and biological characteristics and the outcomes of BCR-ABL1+ ALL according to the different transcripts. Completing the project will take more time and more resources, and we report here the first data set: 497 patients (58.5%) expressed only e13a2, a proportion that was significantly higher than the proportion of e13a2 that was found in newly diagnosed chronic phase CML (37.9%) (Fisher’s exact test, two-sided, p value < 0.0001). This difference cannot be explained by age; indeed the frequency of Ph+ ALL increases with age and is highest in the elderly, where the frequency of e13a2 in CML is the lowest [3, 4, 8]. The finding that the distribution of the different breakpoints in BCR-ABL1 fusion gene is not by chance, being significantly different between CML and Ph+ ALL is of interest. While we are collecting data to compare the characteristics, the response to therapy and the outcomes according to transcript, other important questions, are unanswered. First, why is the distribution different, is the cause related to the cell where the translocation occurs (a multipotent stem cell versus a committed lymphoid progenitor)? Is it due to a different chromatine structure that may influence the position of the breakpoint and the formation of the hybrid gene [9], or to an aberrant activity of the machinery for V(D)J or class switch recombination, or to different DNA-repair mechanisms, or to a microenvironment with a different oxygen tension? Second, what are the biological and clinical consequences of the difference in transcript types, the e13a2 gene being possibly associated with a minor sensitivity to TKIs in CML [8], and at the same time being prevalent in an aggressive leukemia like ALL, that is only temporarily sensitive to TKIs [10–12]? Third, is the transcription efficacy of the two fusion genes identical, or is the turnover of the two mRNAs different, resulting in different amounts of transcript? Fourth, have the two proteins the same leukemogenic potency, have they the same turnover, is the cellular level identical? Fifth, is the sensitivity of the two chimeric proteins to TKIs different? Sixth, is the immunogenicity of the two chimeric transcripts and of the two resulting proteins identical? Of course all these questions need to be addressed also for the more common form of Ph+, BCR-ABL1P190+ ALL (e1a2 or e1a3) [13–15], not forgetting that P190 can also cause CML, although rarely [8]. The answer to these questions may help to optimize the management of Ph+ ALL patients, and also to improve our knowledge on the relationships of genes and gene/protein expression with leukemia. Acknowledgements This work was supported in part by the European Leukemia Net Foundation and by AIRC 5×1000 (21198). The technical assistance of Mrs Chiara Ferri is kindly acknowledged. Compliance with ethical standards Conflict of interest The authors declare that they have no conflict of interest. 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