We show the results of treatment with percutaneous transluminal angioplasty on 38 occasions for vascular access for hemodialysis. Our study includes 22 patients with A-V internal fistulas (Cimino type), 1 autologous saphenous vein shunt and 7 synthetic polytetrafluoro-ethylene shunts. Twenty patients have had a follow-up period over 24 months. Four patients required a second and 2 of them a third transluminal dilatation. Only on two occasions were complications related to the technique. The microscopic findings of the venous wall after percutaneous transluminal angioplasty are shown.
Background: According to WHO-HAEM5, the majority of precursor B-cell acute lymphoblastic leukemia (B-ALL) cases are classified based on cytogenetic testing according to ploidy changes or well-known chromosomal rearrangements. Approximately 30% of patients with B-ALL, however, display none of the major chromosomal abnormalities. This population is classified together as B-other-ALL, a highly diverse subgroup with heterogenous genetic profiles, clinical characteristics, and outcomes. Recent advances in gene expression profiling and sequencing allowed to identify multiple genetic drivers that confer distinct clinical and prognostic features in B-other ALL resulting in better understanding of this subgroup. Among them, B-ALL with specific gene expression signatures (i.e. BCR-ABL1-like and ETV6-RUNX1-like) have been described and included in the classification. However, due to the complex mechanisms underlying the phenotype, standard molecular and cytogenetic workup is not sufficient to make a diagnosis and there is still no universally accepted definition or diagnostic algorithm. Therefore, there is a need to improve our understanding of genetic aberrations driving B-other ALL, screen for unknown lesions, and associate these findings with clinical picture. Aims: Genetic characteristics of pediatric B-other ALL cohort treated in a single center in Poland to evaluate population frequencies of known ALL subtypes, genetic aberrations, and evaluate their associations with outcome and clinical and demographic parameters. Methods: The study included 54 consecutive children (aged 1-18 years) diagnosed in 2014-2022 with B-other ALL, i.e. excluding hyperdiploidy, hypodiploidy, ETV6-RUNX1, BCR-ABL1, TCF3-PBX1, and KMT2A-fusions. In all samples, targeted RNA sequencing using FusionPlex Acute Lymphoblastic Leukemia library preparation kit (ArcherDx) was performed. The samples were sequenced on Illumina MiSeq platform. The data were analyzed using Archer Analysis software v7.0 and aimed to identify fusions, point mutations and expression levels in 81 genes, including fusion transcripts with unknown partners. All novel in-frame fusion transcripts were confirmed with Sanger sequencing. Clinical data was obtained retrospectively from medical records. Results: The cohort included 35% of BCR-ABL1-like cases, 2% ETV6-RUNX-like, and 5,5% PAX5 rearranged (1 fusion, 2 mutations). Mutations activating RAS/RAF-MAPK signaling were observed in 64% of cases and spread across all subtypes. JAK-STAT class aberrations were observed in 37% cases whereas ABL-class aberrations – in 7,2%. Three novel in-frame fusion transcripts were detected, including two lesions affecting cytokine/kinase signaling related to BCR-ABL1-like phenotype (TCOF3/PDGFRB and CD74/PDGFRB) and one affecting DNA repair mechanisms (SSBP2/CHD1). Novel aberrations were associated with poor prognosis. In addition, HOOK3/FGFR1 fusion transcript previously described for other hematological malignancies, but not in ALL, was detected in a single subject. Summary/Conclusion: Our study shows population-based frequencies of novel ALL subtypes, including both recurrent and novel genetic aberrations. This data widens our knowledge on the interplay between molecular aberrations and clinical course of the disease and provides clues for diagnostics optimization. Keywords: B cell acute lymphoblastic leukemia, Gene fusion, Mutation analysis
Background.In recent years, many novel myositis-specific autoantibodies (MSAs) have been identified.However, their links with the pathogenesis and clinical manifestations of inflammatory myopathies remain uncertain. Objectives.To characterize the population of adult dermatomyositis (DM) and polymyositis (PM) patients treated at our center for autoimmune diseases using clinical and laboratory measures. Materials and methods.According to the Bohan and Peter criteria, we retrospectively analyzed patients who fulfilled diagnostic criteria for DM or PM.Myositis-specific autoantibodies and myositis-associated autoantibodies (MAAs) were identified using immunoblot assays.Results.Fifty-one PM (71% women) and 36 DM (67% women) Caucasian patients with a median age of 58 (range: 21-88) years who met the definite or probable diagnostic criteria for myositis were included in the study.Myositis-specific autoantibodies were identified in 63 (72%) patients, whereas MAAs were observed in 43 (49%) of them.Interstitial lung disease (ILD) was characteristic of PM patients (67%, χ 2 with Yates's correction (χ c 2 ) = 13.8078,df = 1, p = 0.0002), being associated with anti-Jo-1 or anti-PL-12 antibodies (fraction comparison test (FCT) 6.4878, p < 0.0001, 6.8354, p = 0.0003, respectively).Interestingly, among patients with anti-MDA5 antibodies (n = 8, 9.2%), all but one had an amyopathic form, with more frequent ILD, skin changes and arthralgias than observed in other patients (FCT 4.7029, p = 0.0228 and p = 7.7986, p = 0.0357, p = 4.7029 and p = 0.0228, respectively).Anti-signal recognition particle (SRP) was strongly associated with the Raynaud's phenomenon (FCT 4. 1144, p = 0.0289) and the highest muscle injury markers (Mann-Whitney U test, z = 2.5293, p = 0.0114).Malignancy was recorded in 14 (16%) patients and was equally common in those with PM and DM.The anti-TIF-1γ was the most frequently related to cancer χ 2 = 14.7691, df = 1, p < 0.0001).The anti-Mi-2α, similarly prevalent in DM and PM, was typically accompanied by skin changes (FCT 7.7986, p = 0.0357) but not ILD (FCT 8.7339, p = 0.0026). Conclusions.Identification of MSAs might help to predict the clinical course of the autoimmune myopathy and malignancy risk.However, these antibodies were absent in about 30% of patients with typical PM or DM manifestations, which encourages further research in this area.
50 patients, 31 male and 19 female with mean age of 45.1 +/- 9.4 years afflicted with blood hypertension (BH) were studied. RDA was performed on all of them as part of the etiological study. The variables evaluated were: systolic blood pressure (SBP), diastolic blood pressure (DBP), therapy index (TI), evolution time of its BH and type of BH (refractory BH, severe BH, mild-moderate BH). The organ affliction was also evaluated (renal function, ECG, presence of cardiomyopathy, vascular disease, and retinopathy. RDA alteration appeared in 16 cases, 1 (10%) in refractory BH group, 8 (28.6%) in severe BH group, and 7 (58.3%) in mild-moderate BH group. A shorter BH evolution period having been observed in patients with altered RDA than in those with normal RDA (3.49 +/- 3.96 years vs 6.93 +/- 4.68 years p = 0.01). We conclude that only this variable is a feature for suspicion of renovascular BH, without an apparent difference between the results of RDA and those of I.V. urography, obtained during the diagnosis screening. The significant differences observed between the mild-moderate BH and the other groups suggests that the clinical suspicion, and not the severity of the BH, is the point which should determine the patients to be renovascularly explored.
Asthma therapy with monoclonal antibodies is a promising and effective approach for those with a severe and refractory type of disease. Although such a targeted therapy is considered to be safe, unusual complications may occur. We present a case of a 45 year-old female patient with severe allergic asthma and chronic spontaneous urticaria, who developed autoimmune polyendocrine syndrome type 2 (APS-2) after 26 months of omalizumab administration. The patient was diagnosed with primary adrenal insufficiency (Addison's disease) and Hashimoto's thyroiditis accompanied by autoimmune atrophic gastritis. According to our knowledge this is the first description of APS-2 that developed in conjunction with omalizumab treatment, although we have no evidence that the observed phenomenon indicated a cause-effect relationship to omalizumab.
Topic: 1. Acute lymphoblastic leukemia - Biology & Translational Research Background: Acute lymphoblastic leukemia (ALL) comprises multiple entities with distinct molecular and clinical features. The latest WHO classification distinguishes several new ALL subtypes, including Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL) - a high-risk subtype characterized by a kinase-activated gene expression profile resulting from multiple diverse genetic alterations. Although Ph-like ALL has been described more than a decade ago and multiple clinical trials have shown that targeted therapy may overcome adverse risk, the identification of Ph-like ALL patients, due to its challenging and not standardized nature, is still not routinely performed outside clinical trials. Due to its rapid turnaround time, cost-effectiveness, and wide availability of both the technology and diagnosticians experienced in immunophenotyping in virtually all hematologic departments, flow cytometry is a perfect candidate for the development of a screening test for Ph-like ALL. The overexpression of cytokine receptor-like factor-2 (CRLF2) has been shown to correlate with CRLF2 rearrangements and has been suggested as a screening tool for the identification of CRLF2-rearranged Ph-like ALL, comprising approximately 50% of Ph-like ALL cases. However, it does not allow to distinguish between CRLF2-overexpressing Ph-like and non–Ph-like ALL, and its use is limited to CRLF2 rearrangements only. To date there is no cytometric method to screen for the remaining half of Ph-like ALL cases. Aims: The study aims to evaluate the expression of a panel of surface and cytoplasmic proteins in ALL blast cells as a surrogate marker of kinase activation enabling routine screening for Ph-like ALL using widely available, multicolor flow cytometry. Methods: Candidate surface and intracellular markers were selected based on the analysis of published expression profiles of ALL patients. The extended panel of surface and intracellular markers included both standard and candidate immunophenotypic markers, i.e. CD45, CD3, CD19, CD34, HLA-DR, phospho-TYR, phospho-STAT3, phospho-STAT5, phospho-CRKL, CRLF2 CD127, CD115, CD25, SOCS1, BLNK, CD97, CD99, CD140A, CD140B, and viability stains (FVS780 and/or 7AAD). For candidate markers, the results were expressed as mean fluorescence intensity (MFI) (for phospho-STAT3, -STAT5 or -CRKL MFI) or as a ratio between respective MFI observed in CD19+ blasts and CD3+ cells that served as internal control (other markers). Clinical data, including classification to ALL subtype, were retrospectively obtained from medical records. For all B-other cases, panel RNA sequencing was performed to allow for classification of new ALL subtypes including Ph-like ALL. Results: Development cohort consisted of 158 bone marrow samples obtained at diagnosis from adult and pediatric ALL patients, including various molecular subtypes of the disease. Except for CRLF2, none of the single markers alone allowed to distinguish new subtypes of ALL. However, multiparameter statistical analysis of the data allowed to build a Ph-like ALL predictor for the identification of Ph-like ALL cases. Prospective analysis of validation cohort patients is ongoing. Summary/Conclusion: We propose a cytometric panel coupled with a statistical model that is capable to identify Ph-like ALL cases at diagnosis in routine clinical settings. Keywords: ALL, Flow cytometry, Acute lymphoblastic leukemia
Patients with systemic sclerosis experience endothelial dysfunction and damage even in the absence of clinical manifestations.To evaluate various methods for assessing the endothelial function for their applicability to clinical practice.Forty-two patients (7 men and 35 women) with systemic sclerosis and 36 controls (11 men and 25 women) matched for age, sex, body mass index, smoking habit, and comorbidities were enrolled in the study. We assessed each participant for typical risk factors for cardiovascular diseases and measured serum levels of vascular cell adhesion molecule-1 (VCAM-1) and thrombomodulin together with flow-mediated dilatation (FMD) of the brachial artery and intima-media thickness (IMT) of the common carotid artery using ultrasonography.Patients with systemic sclerosis did not differ from controls in serum levels of VCAM-1 and thrombomodulin, however, the statistical analysis with adjustment for potential confounders revealed increased levels of thrombomodulin in the patients (p = 0.03). They also had a 45% lower relative increase of FMD (FMD%), and 13% higher IMT (p < 0.01, both, also after adjustment for potential confounders). In a simple regression model, lower FMD% was determined by age (β = -0.57, 95% confidence interval (CI): -0.72 to -0.43) and C-reactive protein levels (β = -0.38, 95% CI: -0.55 to -0.22). Thicker IMT was related to age (β = 0.64, 95% CI: 0.52-0.67), glomerular filtration rate (β = -0.34, 95% CI: -0.5 to -0.18), and blood thrombomodulin levels (β = 0.45, 95% CI: 0.13-0.76).Patients with systemic sclerosis present with endothelial dysfunction which may be detected using ultrasonographic methods. The exact mechanism of observed abnormalities is unknown, but it is possibly related to the chronic inflammation and ischemia-reperfusion injury.
Background: Clinical success of tyrosine kinase inhibitors (TKIs) has transformed chronic myeloid leukemia (CML) into treatable but not curable, truly chronic disease and established treatment-free remission as a new goal for CML therapy. TKI discontinuation is feasible in approximately half of CML patients with stable deep molecular response, but due to the persistence of quiescent leukemic stem cells (LSC) the risk of relapse remains. The observation that peroxisome proliferator-activated receptor γ (PPARγ) agonist, thiazolidinedione (TZD) family, pioglitazone, targets CML-LSCs in vitro has initiated several clinical trials that showed the potential of the combined treatment to allow safe discontinuation of imatinib after concomitant use of pioglitazone. The treatment is well tolerated, its efficacy, however, remains ambiguous. Since it was shown that histone deacetylase inhibitor (HDACi) treatment may also target CML-LSCs in patients receiving tyrosine kinase inhibitors, there is a strong rationale to combine PPARγ agonist and HDACi activities to effectively deplete CML-LSCs and allow for safe treatment discontinuation. To achieve that goal dual PPARγ agonist and HDACi has been designed and synthetized. Here, we present in vitro data on its potential to treat CML. Aims: Evaluation of anti-leukaemic properties of PPARγ agonist/ HDACi dual TZD in combination with TKIs for CML treatment. Methods: A series of double activity TZDs has been designed and synthetized. MTS and XTT cytotoxicity assays in K-562 cell line were used to determine IC50 of all compounds and select a compound with the highest anti-leukaemic activity and the highest potential for combination therapy with TKIs (imatinib and dasatinib). All studies were performed using the selected novel compound (7l) in comparison to pioglitazone. PPARγ activation was measured using Transcription Factor Assay Kit (Abcam), while HDAC activity using Western blot for total H3ac. To elucidate the mechanism of action of the new compound cell cycle analysis, apoptosis assay, growth curve, and clonogenic assay were performed. In addition, high-throughput functional kinase assay using PamGene technology was performed to evaluate kinase activation profile after 7l administration. The results were confirmed with phosphoflow. Results: Addition of novel, double-acting TZD 7l to TKIs significantly decreased clonogenic potential of K-562 cells affecting not only the number but also the size and morphology of the colonies suggesting strong effect on stem-like cells. Similar increase in antileukemic efficacy of TKIs was observed in cytotoxic assays. Moreover, 7l showed stronger synergistic effect with TKIs compared to pioglitazone. Cell cycle arrest in G1/G0 phase was observed, that is concordant with the effects of pioglitazone treatment. Apoptosis assays and growth analysis showed that 7l inhibits proliferation of CML cells without significant increase in cell death. High-throughput kinase activity analysis showed mild inhibition of BCR-ABL signaling in 7l-treated cells (not observed in pioglitazone-treated cells) and inhibition of MAPK, Ras and PI3K-Akt signaling pathways. Summary/Conclusion: Double acting thiazolidinedione 7l with PPARγ ligand and HDACi properties in combination with TKIs significantly reduces clonogenic potential of CML cells and exerts higher antileukemic potential than pioglitazone. Further studies are warranted. Keywords: HDAC inhibitor, Leukemic stem cell, Tyrosine kinase inhibitor, Chronic myeloid leukemia
The aim of the study was to evaluate the clinical phenotypes of glucokinase-maturity-onset diabetes of the young (GCK-MODY) pediatric patients from Southwest Poland and to search for phenotype-genotype correlations.We conducted a retrospective analysis of data on 37 CGK-MODY patients consisting of 21 girls and 16 boys of ages 1.9-20.1 (mean 12.5±5.2) years, treated in our centre in the time period between 2002 and 2013.GCK-MODY carriers were found in a frequency of 3% among 1043 diabetes mellitus (DM) patients and constituted the second most numerous group of DM patients, following type 1 DM, in our centre. The mean age of GCK-MODY diagnosis was 10.4±4.5 years. The findings leading to the diagnosis were impaired fasting glucose (IFG) (15/37), symptoms of hyperglycemia (4/37), and a GCK-MODY family history (18/37). Mean fasting blood glucose level was 6.67±1.64 mmol/L. In the sample, there were patients with normal values (4/37), those with DM (10/37), and IFG (23/37). In OGTT, 120 min glucose level was normal in 8, diabetic in 2, and characteristic for glucose intolerance in 27 of the 37 cases. Twelve of the 37 cases (32%) were identified as GCK-MODY carriers. In the total group, mean C-peptide level was 2.13±0.65 ng/mL and HbA1c was 6.26±0.45% (44.9±-18 mmol/mol). Thirty-two patients had a family history of DM. DM autoantibodies were detected in two patients. The most common mutations were p.Gly318Arg (11/37) and p.Val302Leu (8/37). There was no correlation between type of mutations and plasma glucose levels.The phenotype of GCK-MODY patients may vary from those characteristic for other DM types to an asymptomatic state with normal FG with no correlation with genotype.
Tumor necrosis factor (TNF)-α is a proinflammatory cytokine that plays an important role in the pathogenesis of autoimmune diseases. The aim of the study was to establish an association between TNF-α promoter variability and systemic sclerosis (SSc). The study included 43 SSc patients and 74 controls. Four single nucleotide polymorphisms (rs361525, rs1800629, rs1799724, and rs1799964) located at the promoter of the TNFA gene were genotyped using commercially available TaqMan allelic discrimination assays with real-time PCR. The rs1799724 allele was associated with an increased SSc susceptibility (p = 0.028). In turn, none of the polymorphisms studied were related to the clinical and laboratory parameters of SSc patients, except for a higher prevalence of anti-Ro52 antibodies in the AG rs1800629 genotype in comparison to GG carriers (p = 0.04). Three of four cancer patients had both CT rs1799964 and AG rs361525 genotypes; thus, both of them were related to the increased risk of cancer, as compared to the TT (p = 0.03) and GG carriers (p = 0.0003), respectively. The TNFA C rs1799724 variant is associated with an increased risk of SSc, while the CT rs1799964 and AG rs361525 genotypes might enhance cancer susceptibility in SSc patients, although large observational and experimental studies are needed to verify the above hypothesis.
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