HPLC-UV and GC-MS Methods for Determination of Chlorambucil and Valproic Acid in Plasma for Further Exploring a New Combined Therapy of Chronic Lymphocytic Leukemia.

High performance liquid chromatography with ultra-violet detection (HPLC-UV) and gas chromatography-mass spectrometry (GC-MS) methods were developed and validated for the determination of chlorambucil (CLB) and valproic acid (VPA) in plasma, as a part of experiments on their anticancer activity in chronic lymphocytic leukemia (CLL). CLB was extracted from 250 µL of plasma with methanol, using simple protein precipitation and filtration. Chromatography was carried out on a LiChrospher 100 RP-18 end-capped column using a mobile phase consisting of acetonitrile, water and formic acid, and detection at 258 nm. The lowest limit of detection LLOQ was found to be 0.075 µg/mL, showing sufficient sensitivity in relation to therapeutic concentrations of CLB in plasma. The accuracy was from 94.13% to 101.12%, while the intra- and inter-batch precision was ≤9.46%. For quantitation of VPA, a sensitive GC-MS method was developed involving simple pre-column esterification with methanol and extraction with hexane. Chromatography was achieved on an HP-5MSUI column and monitored by MS with an electron impact ionization and selective ion monitoring mode. Using 250 µL of plasma, the LLOQ was found to be 0.075 µg/mL. The accuracy was from 94.96% to 109.12%, while the intra- and inter-batch precision was ≤6.69%. Thus, both methods fulfilled the requirements of FDA guidelines for the determination of drugs in biological materials.

ACTN3 Genotype in Professional Sport Climbers.

Ginszt, M, Michalak-Wojnowska, M, Gawda, P, Wojcierowska-Litwin, M, Korszen-Pilecka, I, Kusztelak, M, Muda, R, Filip, AA, and Majcher, P. ACTN3 genotype in professional sport climbers. J Strength Cond Res 32(5): 1311-1315, 2018-The functional RR genotype of the alpha-actinin-3 (ACTN3) gene has been reported to be associated with elite sprint/power athlete status. Although large and rapidly increasing number of studies have investigated the associations between the ACTN3 genotypes and athletic performance in various sport disciplines, there is a lack of studies on the genetic predisposition in sport climbing, which was selected to be part of the next Summer Olympic Games in Tokyo 2020 with three subdisciplines ("lead climbing," "speed climbing," and "bouldering"). The aim of the study is to determine the frequency distribution of ACTN3 genotypes and alleles in professional lead climbers and boulderers. 100 professional sport climbers from Poland, Russia, and Austria were divided into 2 equal groups: professional boulderers and professional lead climbers were involved in the study. ACTN3 allele frequencies and genotypes were compared with 100 sedentary controls. Genotypes were determined using polymerase chain reaction-restriction fragment length polymorphism method. The percent distribution of RR genotype in the boulderers was significantly higher than in lead climbers and controls (62 vs. 26%; 33%, respectively; χ = 17.230, p = 0.0017). The frequencies of ACTN3 R allele in boulderers differed significantly from lead climbers and controls (77 vs. 51%; 58%, respectively; χ = 15.721, p = 0.0004). The proportion of the ACTN3 RR genotype is significantly higher in boulderers than in lead climbers and may be related to the specific type of predisposition to this subdiscipline.

Expression of circulating miRNAs associated with lymphocyte differentiation and activation in CLL-another piece in the puzzle.

Expression of microRNAs is altered in cancer. Circulating miRNA level assessed in body fluids commonly reflects their expression in tumor cells. In leukemias, however, both leukemic and nonleukemic cells compose circulating miRNA expression profile of peripheral blood. The latter contribution to extracellular miRNA pool may result in specific microenvironmental signaling, which promotes proliferation and survival. In our study, we used qT-PCR to assay peripheral blood serum of 22 chronic lymphocytic leukemia (CLL) patients for the expression of 84 miRNAs associated with activation and differentiation of B and T lymphocytes. Results were analyzed regarding the most important prognostic factors. We have found that the general expression of examined miRNAs in CLL patients was lower as compared to healthy volunteers. Only miR-34a-5p, miR31-5p, miR-155-5p, miR-150-5p, miR-15a-3p, and miR-29a-3p were expressed on a higher level. Alterations of expression observed in CLL patients involved miRNAs associated both with B and T lymphocyte differentiation and activation. The most important discriminating factors for all functional miRNA groups were trisomy 12, CD38 expression, B2M level, WBC, and NOTCH1 gene mutation. Correlation of expression of miRNAs related to T lymphocytes with prognostic factors proves their supportive function in a leukemic microenvironment. Further studies utilizing a larger test group of patients may warrant the identification of circulating miRNAs that are key players in intercellular interactions and should be considered in the design of microenvironment-targeted therapies.

The incidence of breast cancer in population of young women from Podkarpackie province in 2002-2011.

INTRODUCTION: Breast cancer (BC) in young women of Podkarpackie province accounts for approximately 11.0% of all diagnosed breast tumors. AIM OF THE STUDY: Aim of the study was to assess the trends in incidence of BC among women younger than 44 from Podkarpackie in the years 2002-2011. MATERIAL AND METHODS: 614 cases of malignant BC and 26 cases of cancer in situ were analyzed. The crude and the standardized incidence ratios were estimated; the percentage of histopathologically confirmed cancer cases and the percentage share of registered malignant breast tumors were calculated. The analysis of incidence in individual counties was also presented, as well as the stages of clinical advancement at diagnosis and the methods of treatment. RESULTS: The number of registered cases at 2011 was 73, and it was 37.7% higher as compared to 2002. During the period analyzed, the increase in the crude and the standardized cancer incidence ratios was observed. The percentage share of BC in the examined group increased of 5.4% in 2011. Significant variation in incidence among different counties was observed. The incidence ratios ranged from 65.8 to 93.1/100 000. BC in young women most commonly was diagnosed as locally advanced and over 70% of patients were radically treated. CONCLUSIONS: Even though the progress in diagnostics and treatment has been made, BC in young women is diagnosed later than it should be and at considerably advanced stage. It is relevant to propagate the knowledge among women and health professionals to emphasize that BC may affect young women.

[Epidemiology, prevention and risk morbidity factors for lung cancer]. / Epidemiologia, profilaktyka i czynniki ryzyka zachorowania na raka pluca.

Lung cancer incidence kept increasing dynamically in male population until the late 90s and then there was a sudden drop in the cases and this tendency has been maintained up till now. What seems upsetting, however, is the fact that for female population there is a constant growth in the lung cancer morbidity. Needless to say, Poland still belongs to the countries with high lung cancer incidence and lung cancer mortality. In 2011 the standardized morbidity rate in Poland accounted for 50,0/100 000 in male population and 17,3/100 000 in female population. In Podkarpacie Voivodeship it was 43,6/100 000 for males and 11,8/100 000 for females respectively. Lung cancer incidence and lung cancer mortality seem to increase together with age, and for people 65 and more this type of cancer accounts for approximately 50% of all cancer cases and cancer caused deaths. In spite of various research conducted and great medical progress little can be done to cure lung cancer. The percentage of 5-year survivals increased for males from 10,8% in years 2000-2002 to 11,9% in years 2003-2005, and for females from 15,7% to 16,9%. The main cause of lung cancer is certainly active and passive smoking. It is highly possible that environmental factors are also responsible for lung cancer cases. Among the most devastating are such factors as asbestos, arsenic, aromatic hydrocarbons, individual lifestyle and nutrition, genetic predisposition and finally the pollution, particularly of the air.

Circulating microenvironment of CLL: are nurse-like cells related to tumor-associated macrophages?

B-cell chronic lymphocytic leukemia (B-CLL) is one of the most common hematologic malignancies in Western countries. Accumulation of leukemic lymphocytes in peripheral blood, bone marrow and secondary lymphatic organs of CLL patients is due to decreased apoptosis rather than to increased proliferation. The former is driven by signals from a specific microenvironment, created by stromal cells of mesenchymal origin, follicular dendritic cells, T lymphocytes and others. Nurse-like cells (NLCs) were first described to differentiate from peripheral blood mononuclear cells of CLL patients in vitro, then they have been also found in proliferation centers of their lymphatic tissues. Like tumor-associated macrophages (TAMs) in solid tumors, nurse-like cells promote survival of CLL lymphocytes. NLC gene expression patterns suggest their similarity to TAMs and differ between patients depending on ZAP70 protein expression status. NLC number in vitro corresponds with CD14 expressing cell count and beta-2-microglobulin serum level, and positively correlates with leukemic lymphocyte viability. As NLCs strongly express genes for adhesion molecules and secrete chemokines of antiapoptotic activity, they should be considered as a target for anti-microenvironment therapy of this incurable disease.

[Biogenesis of microRNAs and their role in the development and course of selected hematologic disorders]. / Biogeneza czasteczek mikroRNA oraz ich znaczenie w powstawaniu i przebiegu wybranych zaburzen hematologicznych.

MicroRNAs (miRNAs) are a group of small molecules, about twenty nucleotides in length. They are involved in the regulation of gene expression mainly at a posttranscriptional level. This function depends on their complementarity to the 3'UTR regions of mRNAs. MicroRNAs are essential for proper development and functioning of the organism. They are so important because of their participation in such processes as angiogenesis, apoptosis, cell cycle control and oncogenesis. Over thirty percent of human genes are controlled by miRNAs. This indicates the great importance of these molecules. Alterations of numerous cellular processes can be caused by the dysregulation of miRNA expression. Such disturbances are observed in cancer cells, and signatures of microRNA expression are specific to particular types of cancer. It is suggested that miRNAs may serve as diagnostic and prognostic factors in oncologic and hematooncologic disorders. The expression of specific miRNAs can indicate benign or aggressive course of disease. The overall survival or time to treatment are also possible to estimate based on the microRNA expression profile. Knowledge about changes in miRNA expression observed in leukemia patients may enable the selection of appropriate individual therapy. Recent reports indicate that various hematooncologic disorders may be well characterized by microRNAs circulating in plasma or serum. It is of great potential importance, considering the availability of material for analysis, simplicity of diagnostic procedures and shortening of time required to conduct them.

Inside the chronic lymphocytic leukemia cell: miRNA and chromosomal aberrations.

Alterations in microRNA (miRNA/miRs) expression are associated with the occurrence and course of human diseases, including chronic lymphocytic leukemia (CLL). Expression of miRNAs may vary among patients with CLL in different cytogenetic risk groups. The present study assessed the expression levels of the following miRNAs in 35 patients with CLL: hsa­miR­15a, ­16­1, ­29a, ­29c, ­34a, ­34b, ­155, ­181a, ­181b, ­221, ­222 and ­223. Fluorescent in situ hybridization (FISH) analysis was performed for 13q14d, 17p13 and 11q22 deletions and chromosome 12 trisomy. Significantly higher expression levels of miR­181a, ­221 and ­223 were observed in the group at low risk of disease progression (stage 0) compared with the group with high risk of CLL progression (P=0.036, P=0.019 and P=0.038, respectively). The present study revealed that the expression levels of miRNA­181b and miRNA­223 were significantly higher in the group of patients without D13S319 deletion (P=0.039 and P=0.037, respectively). Moreover, the expression levels of miR­15a and miRNA­29c were demonstrated to be significantly higher in the group of patients with CLL who had a tumor protein p53 deletion, identified by FISH, compared with patients without this lesion (P=0.047, P=0.03 respectively). Based on receiver operating characteristic curve analysis, the present study revealed that miR­181a, ­221 and ­223 expression was able to distinguish low and high risk of CLL progression in patients. Among the tested miRNAs, miRNA­181a, ­221 and ­223 were indicated to have the greatest diagnostic potential in CLL.

[Molecular and cytogenetic prognostic factors in acute myeloid leukemia (AML)]. / Molekularne i cytogenetyczne czynniki prognostyczne w ostrej bialaczce szpikowej (OBS).

Acute myeloid leukemia (AML) constitutes a group of diseases that are very heterogeneous with regard to clinical course, response to therapy as well as cytogenetic aberrations and gene mutations. Such lesions are of prognostic value. Patients with t(8;21), inv(16)/t(16;16) or t(15;17) have a favorable prognosis. Patients with normal karyotype and aberrations +6, +8 ­Y, t(9;11) and del(12p) are classified in the group of intermediate prognosis. In the case of patients with complex karyotype or with the aberrations inv(3)/t(3;3), t(6;9), ­5, ­7, del(5q), del(7q) and 11q23 rearrangements, the prognosis is poor. Unfavorable molecular factors include C-MYC amplification, MLL amplification and rearrangement, FLT3-ITD, WT1 mutation and overexpression of BAALC, ERG or MN1. The changes in miRNA expression may also be important for AML prognosis. That is why the cases with normal karyotype (CN-AML) and cases with complex aberrations remain to be better characterized at the genetic level. Array technology enables the analysis of genomic DNA and gene expression. This approach may be used in the search for new prognostic and predictive markers in AML.

Anticonvulsant valproic acid and other short-chain fatty acids as novel anticancer therapeutics: Possibilities and challenges.

Results from numerous pre-clinical studies suggest that a well known anticonvulsant drug valproic acid (VPA) and other short-chain fatty acids (SCFAs) cause significant inhibition of cancer cell proliferation by modulating multiple signaling pathways. First of all, they act as histone deacetylase (HDAC) inhibitors (HDIs), being involved in the epigenetic regulation of gene expression. Afterward, VPA is shown to induce apoptosis and cell differentiation, as well as regulate Notch signaling. Moreover, it up-regulates the expression of certain G protein-coupled receptors (GPCRs), which are involved in various signaling pathways associated with cancer. As a consequence, some pre-clinical and clinical trials were carried out to estimate anticancer effectiveness of VPA, in monotherapy and in new drug combinations, while other SCFAs were tested in pre-clinical studies. The present manuscript summarizes the most important information from the literature about their potent anticancer activities to show some future perspectives related to epigenetic therapy.

Examination of clonal evolution in chronic lymphocytic leukemia.

Chronic lymphocytic leukemia (CLL) is one of the most frequent lymphoproliferative diseases. CLL is characterized by unusual heterogeneity, which probably reflects its biological and genetic lack of homogeneity. Clonal chromosome aberrations belong to the most important prognostic and predictive factors in CLL. This research was aimed at observing clonal evolution in CLL at the chromosomal level, and assessing its clinical significance in relation to selected prognostic factors. The study involved 72 untreated patients with CLL. The preliminary investigations using cytogenetic banding analysis (CBA) and FISH were performed at the time of diagnosis, and again after about 24 months to observe clonal changes (clonal evolution). In addition, other parameters were evaluated, i.e., the expression of ZAP-70 kinase, CD38 antigen, and the mutation statuses of IGVH and NOTCH1 genes. Classic prognostic factors, i.e., categorized ZAP70 and CD38 expressions as well as mutations in IGVH and NOTCH1 genes did not influence the course of clonal evolution in the examined group of patients. Clonal evolution was detected in 45.8% of patients by means of CBA, and in 19.4% patients with FISH. Analysis of chromosomal aberrations in the examined group of patients showed that the incidence of 17p deletions and translocations in karyotypes has a negative impact on overall survival. CE was found to be a risk factor for the occurrence of disease progression (OR = 2.22). Our observations indicate that combined CBA and FISH are the most optimal techniques for monitoring clonal evolution in the course of CLL.

The Association of GSTT1, GSTM1, and TNF-α Polymorphisms With the Risk and Outcome in Multiple Myeloma.

Oxidative stress and systemic inflammation are closely linked with increased risk of cancer development. Tumor necrosis factor alpha (TNF-α) is one of the pro-inflammatory cytokines. Glutathione S-transferases (GSTs) are enzymes involved in oxidative stress handling. Polymorphisms of genes encoding mentioned molecules may potentially influence the risk and the outcome in neoplastic diseases. Multiple myeloma (MM) is a hematological malignancy characterized by clonal, atypical plasma cell proliferation. In the present study we investigated the association of deletion polymorphisms in GSTT1/GSTM1 genes and single nucleotide polymorphisms (SNPs) in the TNF-α gene at positions -308/-238 with the risk and outcome in MM and sensitivity to bortezomib under in vitro conditions. One hundred newly diagnosed MM patients and 100 healthy blood donors were genotyped by means of multiplex PCR (for GSTs) and PCR-RFLP (for TNF-α). In a subgroup of 50 MM patients, bone marrow cells were treated with bortezomib in vitro. Patients with -238GA+AA or GSTT1-null genotypes had 2.0 (p = 0.002) or 2.29 (p = 0.013) fold increased risk of MM. The interaction effects and risk of MM were observed in GSTT1/GSTM1-null (OR = 2.82, p = 0.018), -308/-238GA+AA (OR = 5.63, p < 0.001), as well as in all combinations of -308 with GSTs. The -308/-238GA+AA genotypes in comparison to GG were associated with earlier MM onset-61.14 vs. 66.86 years (p = 0.009) and 61.72 vs. 66.52 years (p = 0.035), respectively. Patients with GSTM1-present had shorter progression-free-survival (15.17 vs. 26.81 months, p = 0.003) and overall-survival (22.79 vs. 34.81 months, p = 0.039) compared with GSTM1-null. We did not observe relationship between response rate and studied polymorphisms. The in vitro study revealed significantly higher number of apoptotic cells at 12 nM of bortezomib in GSTT1-present, GSTM1-null/present, -308GG and -238GG/GA+AA genotypes. Our findings comprise large analysis of studied polymorphisms in MM.

ACE Insertion/Deletion Polymorphism (rs4646994) Is Associated With the Increased Risk of Multiple Myeloma.

Introduction: The insertion (I allele) deletion (D allele) polymorphism of ACE gene (rs4646994) may influence the etiopathogenesis of multiple myeloma (MM). ACE gene is expressed in bone marrow cells and encodes angiotensin converting enzyme (ACE). It converts angiotensin I to active peptide angiotensin II, which stimulates proliferation of hematopoietic stem cells. This suggests possible association of ACE I/D gene polymorphism with MM. The aim of our study was to check possible impact of this polymorphism on risk of development and outcome of MM, as well as, sensitivity to bortezomib in cell cultures derived from MM patients. Objects and Methods: Genomic DNA from 98 newly diagnosed MM patients and 100 healthy blood donors were analyzed by PCR method. Chromosomal aberrations were detected by use of cIg-FISH. In a subgroup of 40 MM patients nucleated bone marrow cells were treated with bortezomib in vitro. Results: The Hardy-Weinberg equilibrium test showed that genotypic frequencies diverged significantly from the equilibrium. The differences between I and D allele frequencies in control and study population were significant (p = 0.046). We observed the association between DD genotype and more than 2-fold risk of MM - OR = 2.69; p < 0.0001. We did not detect any significant differences among studied genotypes regarding clinical and laboratory parameters. Moreover, we did not observe the association between survival of MM patients and I/D genotypes. Bortezomib increased number of apoptotic and necrotic cells, but the only statistically significant differences were observed in the number of viable cells at 1 nM between ID and DD genotypes (p = 0.026). Conclusion: Presented results confirmed the significant relationship between ACE (I/D) polymorphism and risk of MM development. We did not observe the association of ACE I/D polymorphism with disease outcome and bortezomib in vitro sensitivity.

Chromosome Preparation for Chronic Lymphoid Malignancies.

Conventional cytogenetics is invariably one of the most important methods used in diagnostics of chronic lymphoproliferations. It complements fluorescence in situ hybridization (FISH) and molecular analysis. Presence of particular chromosomal alterations in chronic lymphocytic leukemia enables patients' stratification into appropriate cytogenetic risk groups and influences treatment decisions. In other non-Hodgkin lymphomas cytogenetic analyses are employed also in minimal residual disease assessment.As lymphocytes in chronic lymphoid malignancies are characterized by low proliferation rate in vitro, it is critical to induce their division in the culture properly. Here, we describe methods of lymphocyte isolation from patient's samples, conditions of cell culture, and the most commonly used mitogens for B- and T-lymphocytes in hemato-oncologic analyses.

Detection of chromosomal changes in chronic lymphocytic leukemia using classical cytogenetic methods and FISH: application of rich mitogen mixtures for lymphocyte cultures.

One of the research methods of prognostic value in chronic lymphocytic leukemia (CLL) is cytogenetic analysis. This method requires the presence of appropriate B-cell mitogens in cultures in order to obtain a high mitotic index. The aim of our research was to determine the most effective methods of in vitro B-cell stimulation to maximize the number of metaphases from peripheral blood cells of patients with CLL for classical cytogenetic examination, and then to correlate the results with those obtained using fluorescence in situ hybridization (FISH). The study group involved 50 consecutive patients with CLL. Cell cultures were maintained with the basic composition of culture medium and addition of respective stimulators. We used the following stimulators: Pokeweed Mitogen (PWM), 12-O-tetradecanoylphorbol 13-acetate (TPA), ionophore, lipopolysaccharide (LPS), and CpG-oligonucleotide DSP30. We received the highest mitotic index when using the mixture of PWM+TPA+I+DSP30. With classical cytogenetic tests using banding techniques, numerical and structural aberrations of chromosomes were detected in 46 patients, and no change was found in only four patients. Test results clearly confirmed the legitimacy of using cell cultures enriched with the mixture of cell stimulators and combining classical cytogenetic techniques with the FISH technique in later patient diagnosing.

Examination of the FLT3 and NPM1 mutational status in patients with acute myeloid leukemia from southeastern Poland.

INTRODUCTION: Acute myeloid leukemia (AML) is a genetically heterogeneous disease at both the cytogenetic and molecular levels. In AML cells many chromosomal aberrations are observed, some of them being characteristic of a particular subtype of patients, and others being less significant. Besides chromosomal abnormalities, the leukemic cells can have a variety of mutations involving individual genes. The aim of this work was to investigate the frequencies of molecular alterations with the focus on FLT3-ITD and NPM1 mutations in AML patients of different age groups living in a southeastern region of Poland. MATERIAL AND METHODS: The study group comprised 50 consecutive AML patients. We analyzed bone marrow samples by conventional cytogenetics. Cytogenetic evaluation in selected cases was complemented by the FISH technique. The internal tandem mutation in the FLT3 gene was identified using polymerase chain reaction (PCR), and the NPM1 mutation was assessed by direct nucleotide sequencing. RESULTS: The studies using classical cytogenetics showed chromosomal aberrations in 32 (64%) patients. In 18 cases no changes in the karyotype were found by conventional karyotyping. FLT3-ITD mutation was detected in 4 (8%) patients and mutation of NPM1 in 3 patients with AML (6%). CONCLUSIONS: The incidence of both mutations in our study group was lower than described elsewhere. We have confirmed that FLT3-ITD occurred more commonly in older patients and it was associated with shorter overall survival. By contrast, mutation of exon 12 of the NPM1 gene seems to be a good prognostic factor in AML patients with normal karyotype.

Guilty bystanders: nurse-like cells as a model of microenvironmental support for leukemic lymphocytes.

B-cell chronic lymphocytic leukemia (B-CLL) is one of the most common leukemias among the elderly and, despite many efforts, still stays incurable. Recent studies point to the microenvironment as the critical factor providing leukemic lymphocytes with pro-survival signals. Thus, the neighboring cells appear to be a perfect target for antileukemic therapy. Nurse-like cells (NLCs) largely contribute to CLL microenvironmental support. We developed the CLL lymphocyte/NLC co-culture model for the investigation of microenvironmental interactions. Viability and apoptosis were investigated in CLL lymphocytes treated with dexamethasone (DEX) and chlorambucil (CLB), with and without NLCs' support. For the first time, the capacity of DEX and CLB to affect NLCs viability was also evaluated. Apoptosis-associated gene expression profiles of leukemic lymphocytes ex vivo and cultured with NLCs were assessed by expression arrays. CLL lymphocytes escaped spontaneous apoptosis for several months when cultured with NLCs. The presence of NLCs significantly reduced apoptosis induced with DEX and CLB (p < 0.001; p = 0.012, respectively), and their protective effect was more evident than the effect of recombinant SDF1. Both DEX and CLB also decreased NLCs viability, but to a lesser extent (mean viability in DEX-treated cultures was 37.79% in NLCs compared to 29.24% in lymphocytes). NLCs induced the expression of important anti-apoptotic genes in cultured CLL lymphocytes; median expression of BCL2, SURVIVIN, BCL2A1, and XIAP was significantly higher as compared to ex vivo status. The CLL lymphocyte/NLC co-culture makes up the convenient and close to the natural-state model for studying the relationship between leukemic cells and the microenvironment. Direct cell-to-cell contact with NLCs increases the expression of anti-apoptotic genes in CLL lymphocytes, thus protecting them against induced apoptosis. As the effect of antileukemic drugs is not so apparent in NLCs, the combined therapy targeted at both lymphocytes and the microenvironment should be considered for CLL patients. Simultaneous aiming at the disruption of several different signaling pathways and/or anti-apoptotic proteins may further improve treatment efficiency.

Steatocystoma multiplex with hair shaft abnormalities.

Steatocystoma multiplex (SM) is an unusual benign disorder of the pilosebaceous duct characterized by multiple cysts with little or no nail and hair involvement. We report a 30-year-old woman with multiple cystic nodules located on the neck, axillae and forearms as well as patchy scalp alopecia. Histopathological examination of the lesions was diagnostic of SM. Trichoscopy revealed pili torti and pili canaliculi. This patient represents an unusual clinical presentation of SM because of the presence of hair abnormalities.

New boys in town: prognostic role of SF3B1, NOTCH1 and other cryptic alterations in chronic lymphocytic leukemia and how it works.

B-cell chronic lymphocytic leukemia (B-CLL) is one of the most common leukemias of the elderly. To date, although many prognostic factors are known, none are universal or easily accessible thus allowing for the stratification of patients to slow-go and aggressive-course groups. Recent studies have identified new recurrent mutations in CLL cells, including mutation of the gene encoding one of the spliceosome subunits, SF3B1, mutation or rearrangement of NOTCH1, a gene of well-known tumorigenesis association, and disruption of BIRC3, a member of the inhibitors of apoptosis (IAP) family. This article presents the current state-of-the-art findings concerning the prognostic significance of these new alterations, as well as an explanation of the mechanisms underlying their biological impact on CLL lymphocytes.