We studied five patients with hairycellleukemia (HCL) diagnosed within the last ten years at the Department of Hematology of Universidade Federal de São Paulo - Escola Paulista de Medicina. Our purpose was to analyze the value of transmission electron microscopy (TEM) by comparing this method with the conventional ones. At diagnosis, patients presented weight loss, spleen enlargement and hairy cells (HC) in peripheral blood and bone marrow slides. HC was characterized by morphology and tartrate test resistance in the acid phosphatase reaction (TRAP). At the evaluation time, the amount of HC ranged from 1% to 85% of WBC count. All patients, except two, had phenotype B. In these last two, TRAP as well as phenotype B could not be documented due to low HC numbers in their exams. Cytoplasmatic projections and the absence of lamellar ribosomic complex were the most frequent ultrastructural findings, even in those patients with the lowest HC numbers. Based on these features, TEM is an efficient method for searching for HC at HCL diagnosis and during the course of the disease.
Approximately 20 percent of patients with Sweet syndrome have an associated cancer . Hematologic malignancies, most commonly acute myelogenous leukemia, account for 85 percent of the associated malignancies [1, 2, 3]. The most common solid tumors are those of the genitourinary tract . We report the case of a patient with Sweet syndrome as the presenting symptom of hairycellleukemia.
When the patient returned to the hospital of the Uni- versidade Federal de Santa Catarina (HU-UFSC) at the end of treatment, the myelogram was hypercellular for his age, but this alteration has no clinical relevance to his condition. The presence of 1.1% of cells with a phenotype similar to that found at diagnosis, as evidenced by immunophenotyping of a bone marrow aspirate, indicates the presence of mini- mal residual disease (MRD). MRD is present in many patients treated for hairycellleukemia; however, these patients may have long-term survival. Treatment with Rituximab is used to achieve complete remission, although the response is depend- ent on how much bone marrow is involved and on the characteristics of the individual. 8
Other entities that might have formed diferential diagno- ses were hairy-cellleukemia and hairy-cellleukemia variant. However, the clinical data in association with morphological and immunophenotyping data made it possible to safely rule out diagnoses of hairy-cellleukemia (absence of pancytopenia, no cells with the classical morphology of “hairycell” and immuno- phenotyping not characterized by strong expression of CD11c, CD25, CD103 and CD123 antigens) or hairy-cellleukemia vari- ant (no lymphocytes with morphology characterized by the pres- ence of central vesicular nucleolus, cytoplasm similar to hairycell and absence of strong expression of CD11c and CD20 antigens). hus, integration of all these indings, i.e. lymphoprolifera- tive malignancy of small B cells, difuse iniltration of the spleen and primary from splenic red pulp, with exclusion criteria for “hairycell” and its variant form, enabled the deinitive accurate diagnosis of splenic difuse red-pulp small B-cell lymphoma in both cases reported here.
In addition, PTB presented a potent cytostatic effect on the leukemiacell line (K-562, GI 50 = 0.91 g/ml) (Table 2), which is in agreement with the results described by Smolarz et al. (2006). PTB also inhibited endothelial cell proliferation, probably by promoting membrane cell depolarization, suggesting a potential antiangio- genic effect (Siekmann et al., 2013). Due to its lipolyphilicity (Log P = 3.1), PTB in rats showed high absorption with a maximum concentration time of about 2 h. A large volume of distribution has been observed, and the metabolism appears to be essen- tially hepatic. Based on clearance values this compound is mainly excreted via non-renal, with serum half-life of approximately 7 h (Sayre et al., 2012, 2015).
flavopiridol alone resulted in 10% and 17% subG1 cells, respectively (Figure 6C). In contrast, seliciclib-resistant ARH77 cells did not respond to low concentrations of seliciclib and flavopiridol, demonstrating only a mild increase in subG1 population following combined treatment (2.7% in control, 6% in seliciclib- and 5.5% in flavopiridol-treated cells vs. 11% in the combined protocol). Notably, elevated dose of flavopiridol (500 ng/ml) increased the apoptosis of ARH77 cells, yielding 21% of subG1 population when applied alone and 35% of subG1 cells in combination with 10 m M of seliciclib (data not show). Moreover, by combining flavopiridol with seliciclib we were able to show effective decrease in CCND1 and CCNE1 expression, evaluated by immunoblotting (Figure 6D). The most prominent effect of the combined treatment on CCND1 expression was detected in ARH77 and NCI H929 cells. Importantly, flavopiridol treatment efficiently decreased the basal and seliciclib-induced levels of CCNE1 in MM cells. These results may explain the mechanism of the observed increased toxicity induced by the combination of seliciclib with flavopiridol. All together, these data Figure 4. Seliciclib downregulates MCL1 expression in hMMCLs. (A) The indicated multiple myeloma cell lines in logarithmic growth phase were extracted and subjected to immunoblotting, utilizing MCL1 antibodies. In all experiments CDK2 expression serves as an internal loading control. Experiments were performed at least 3 times and one representative result is presented. (B–C) MCL1 downregulation by seliciclib. Cells were extracted and subjected to immunoblotting. (B) Various hMMCLs were incubated in the presence of seliciclib 50 mM or DMSO for 6 hours and the level of MCL1 expression was analyzed. (C) RPMI8226, CAG and NCI H929 cells were incubated in the absence or presence of seliciclib 50 mM for the indicated time points, or in the presence of increasing concentrations of seliciclib for 8 hours. (D) Reduction in MCL1 phosphorylation by seliciclib. Cells were treated with 50 mM or DMSO for 8 hours and the levels of total and phosphorylated MCL1 were analyzed by immunoblotting using specific antibodies. b-actin was used to confirm equal protein loading. (E) CAG cells were incubated in the absence or presence of seliciclib or MG132 (10 mM) exclusively or combined. MCL1 level of expression was verified by immunoblotting. b-actin was used to confirm equal protein loading.
3-(Thiophen-2-yl)-4,5-dihydro-1H-pyrazole-1-carboximidamides were efficiently prepared through a cyclocondensation of thiophenylchalcones with aminoguanidine hydrochloride under ultrasonic conditions in the presence of KOH and ethanol as a green solvent in short reaction times (15-35 min) and good yields (62-95%). All compounds produced were evaluated against the human Jurkat and RS4;11 acute lymphoblastic leukemiacell lines of T- and B-cell origin, respectively, and the K562 myelogenous leukemiacell line. Six compounds presented half maximal inhibitory concentration (IC 50 ) values around 15 µmol L -1 and five compounds presented IC 50 values around
Cumulative reports reveal the protean nature of NOTCH signaling in the maintenance of normal and malignant hemato- poiesis [13,24–27,37,38]. While Notch2 signaling regulates re- generation of mouse long-term HSC, ligand-driven NOTCH1 activation induces human hematopoietic progenitor expansion and differentiation [28,39]. Ligand binding to the NOTCH1 extracellular domain activates ADAM family metalloprotease and c-secretase complex-mediated cleavage and intracellular release of the NOTCH1 intracellular domain (ICN1). Subsequently, nuclear translocation of ICN1 followed by engagement of transcriptional activators such as CBF1/Su(H)/Lag2 (CSL) and mastermind-like (MAML) sets the stage for NOTCH1 target gene transcription. Conversely, activation of NOTCH1 signaling through gain-of- function mutations in NOTCH1, first described in T-ALL , or loss-of-function mutations in NOTCH1 regulators, such as FBXW7 and NUMB, has been linked to therapeutic recalcitrance of hematologic malignancies [40,41]. Chronic antagonism of both NOTCH1 and NOTCH2 processing with small molecule inhibitors of the c-secretase complex has been associated with loss of intestinal crypt progenitor cells, thereby providing the impetus for development of selective NOTCH1 inhibitors . Recent pre-clinical studies demonstrate that inhibition of NOTCH1 using synthetic stapled peptides or monoclonal antibody-mediated strategies effectively decreases T-ALL cell line growth [22,23]. However, the consequences of selective NOTCH1 inhibition for normal hematopoietic progenitor and patient- derived T-ALL LIC survival and self-renewal have been unclear.
shown). Therefore we assessed whether FK866 would affect T cell viability and whether resting and activated T lymphocytes would behave differently in terms of susceptibility to this drug. We found that FK866 was toxic to PBLs when these were concomitantly activated with mitogens [phytohematoagglutinin- P (PHA), or concanavalin A (Con A)], while unstimulated cells were mostly unaffected (Figure 1B). Similarly, activation with allogeneic DCs also sensitized PBLs to FK866-induced cell demise while unstimulated PBLs were less affected (Figure 1C, D). As detected after a five-day treatment, FK866-induced cell death in activated PBLs was concentration dependent, with EC50s comprised between 1 and 10 nM, and typically reached a plateau starting from 30 nM (Figure 1B, C). In resting PBLs, FK866 EC50 was never reached in the concentration range we used. In experiments where FK866 treatment (33 nM) was extended for up to 12 days, unstimulated PBLs were still .80% viable indicating that resting T cells are mostly resistant to FK866-induced cell death (data not shown). Similarly, FK866
promoter demethylation (Fig. 4). A detailed analysis of the XAGE1D promoter showed that this promoter is 95% hypermethylated in the untreated HL-60 cell line. A 40% reduction of methylation was induced by both drugs, however 5-Aza-CdR caused approximately 5 times higher expression than 5-Aza-CR at both time points. No significant increase in expression from day 2 to day 8 was seen in this cell line. In T24 cells, 5-Aza-CdR also led to a higher expression level than did 5-Aza-CR. Hardly any expression was induced on day 2 by 5- Aza-CR, and a higher expression level was achieved on day 8 for both drugs (Fig. 4). Similar results were obtained by detailed RT-qPCR analyses of different CTAs (supplementary Fig. S4). Since not all CTAs have a CpG island in their promoters, we speculated whether other factors might have induced the high expression of CTAs. Interestingly, the CTCF paralogue CTCFL/BORIS, that can reactivate the transcription of other CTAs , was also upregulated by both drugs in both cell lines. This gene has CpG islands in 2 of its 3 promoters and 5- Aza-CdR has previously been shown to strongly upregulate transcription by mechanisms both dependent and independent of DNA methylation . In HL-60 cells CTCFL/BORIS showed significant upregulation already on day 2, and its expression coincided with CTA upregulation. CTCFL/BORIS is believed to antagonize CTCF and in this cell line, CTCF was downregulated at each time point by both drugs. Only a minor fraction of the CTAs was upregulated by 5-Aza-CR on day 2 with no upregulation of CTCFL/BORIS in T24 cells. More upregulation of both CTCFL/BORIS and other CTAs was seen by 5-Aza-CdR. On day 8 both drugs showed high upregulation of a large proportion of CTAs including CTCFL/BORIS (Fig. 3).
15. Rossi D, Sozzi E, Puma A, De Paoli L, Rasi S, Spina V, et al. The prognosis of clinical monoclonal B cell lymphocytosis differs from prognosis of Rai 0 chronic lymphocytic leukaemia and is recapitulated by biological risk factors. Br J Haematol. 2009;146(1):64–75.
Human T-cell lymphotropic virus, type I (HTLV-I) is associated to a neoplastic disorder (Adult T-cellLeukemia and Lymphoma-ATLL), to a characteristic uveitis and a neurological chronic disease, Tropical Spastic Paraparesis or HTLV-I Associated Myelopathy (TSP/HAM). This condition is a chronic myelopathy without spontaneous remission, characterized by a slowly progressive paraparesis, affecting mainly the pyramidal tracts and associated to a variable sphincter disturbance and abnormalities of the sensory
In the present study, we demonstrated that when BV and palladium (II) complex were consumed simultan- eously, the combination of 1 μg/mL BV with 0.85 μM Pd (II) complex induces MOLT-4-cell apoptosis in a caspase-3-dependent manner. Orsolic , while investi- gating cytotoxic effects of bee venom applied alone or in combination with the DNA-damaging drug bleomycin on HeLa and V79 cells, found that bleomycin caused a dose-dependent decrease in cell survival. When used with a non-lethal dose of the BV, its lethal effect was po- tentiated. The author inferred that BV, by preventing re- pair of damaged DNA, increases bleomycin lethality and inhibited recovery from bleomycin-induced damage . Because DNA is the main target of palladium metal- based complexes, we may conclude that BV is able to potentiate the lethality effect of [Pd (bpy)(Pi-Pydtc)]NO 3
In addition to a role for hairy in refinement of the terminal cell fate through regulation of bnl expression in muscle cells, we also Figure10. Ectopic expression of bnl in the mesoderm phenocopies hairy mutant phenotype by specifying extra terminal cells. In stage 13 wild-type embryos (A and A’), bnl RNA is expressed in clusters surrounding the migrating trachea (A, arrow) and to a lesser extent in the circular visceral mesoderm (A’, arrowhead). In stage 13 embryos where bnl is overexpressed with twi-GAL4 mef2-GAL4 (B), bnl RNA is expressed in the somatic mesoderm (B, arrowhead). In stage 13 embryos where bnl is expressed with 5053-GAL4 (C and C’), bnl RNA is expressed in clusters close to the tracheal cells (C, arrow) and in longitudinal muscle founder cells of the visceral mesoderm (C’ arrowheads). In stage 16 embryos expressing bnl with 5053-GAL4 (D), bnl RNA is expressed in the ventral oblique lateral body wall muscles (D, arrowhead). In wild-type embryos (E), a single terminal cell (TC) is specified at the tip of each dorsal branch (E, arrow). In embryos overexpressing bnl in the entire mesoderm with twi-GAL4 mef2-GAL4 (F) or in a subset of the mesoderm with 5053-GAL4 (G), extra TCs are specified in the dorsal branches (F and G, arrows). In wild-type embryos (H), a distinct number of TCs are specified in the lateral trunks (H, arrows) and the ganglionic branch (H, arrowhead) whereas in embryos expressing bnl in the mesoderm with either twi-GAL4 mef2-GAL4 (I) or 5053-GAL4 (J) extra TCs in the lateral trunks (I and J, arrows) and ganglionic branches (J, arrowheads) are specified. Embryos in A–D were hybridized in situ with bnl RNA. Embryos in E–J were stained for DSRF (red) and 2A12 (green). All panels shown are lateral views except panel G which is a dorsal-lateral view. Scale bar in E represents 10 mm.
Six days after discharge, the patient re-presented with worsening of symptoms. He continued to describe nausea, vomiting, decreased PO intake, and progressive fatigue. Since discharge, he had lost another 10 pounds. Admission laboratory data revealed a white blood cell count of 17 320/ cm 3 (56% neutrophils, 17% lymphocytes, 10% reactive lymphocytes, 2% eosinophils), creatinine of 2.6 mg/dL, total bilirubin 2.6 mg/dL, aspartate aminotransferase 225 IU/L, alanine aminotransferase 869 IU/L, and alkaline phos- phatase 191 IU/L. Given the persistent abnormalities in his liver tests, a liver biopsy was performed and revealed mod- est hepatocyte apoptosis and scattered portal and lobular inflammatory cells (primarily lymphocytes) felt to be con- sistent with drug-induced or viral hepatitis (Figure 1). Urine studies revealed a fractional excretion of sodium of 2.2%, suggesting intrinsic renal disease. Urinalysis with micros- copy was benign with no evidence of cellular casts, dysmor- phic red blood cells, white blood cells, or crystals. A kidney biopsy was performed to determine the cause of his acute kidney injury as the renal function did not improve with aggressive volume expansion. Two days after kidney biopsy, he developed pain in his left lower extremity, which on ultrasound was found to be caused by an extensive deep vein thrombosis encompassing the posterior tibial vein extending proximal up to the common femoral vein. Given his apparent hypercoagulable state, weight loss, and periph- eral lymphocytosis, a hematologic malignancy was consid- ered. Uric acid 12.3 mg/dL, lactate dehydrogenase 290 IU/L, phosphorus 5 mg/dL, and potassium 4.3 mmol/L. Peripheral blood flow cytometry revealed lymphoblasts, and bone marrow biopsy revealed 78% blasts with markers consistent with B-cell acute lymphoblastic leukemia (Figure 2). In retrospect, the atypical lymphocytes from the initial peripheral smear were actually lymphoblasts, although the blood smear was not available for re-review after the leuke- mia diagnosis. He was started on allopurinol and transferred to the inpatient malignant hematology service where he was started on induction chemotherapy.
Anyway, apoptosis is not the only mechanism of tubulin inhibitor- induced cell death. Several studies have described a form of cell death called mitotic catastrophe. Mitotic catastrophe is an event in which a cell is destroyed during mitosis. It occurs because of an attempt at ab- errant chromosome segregation in mitosis or as a result of DNA dam- age (Castedo et al., 2004; Mollinedo and Gajate, 2003). It has been reported that tubulin-binding agents induce cell death through a mitotic catastrophe (Kanthou et al., 2004; Nabha et al., 2002). Interestingly, Magalhães et al. (2011b) showed that PHT also is able to induce an accu- mulation of metaphase cells on cultured lymphocytes. Moreover, a considerable increase in the frequency of chromosome aberrations was found in cells exposed to PHT. This genotoxic effect can be important as an alternative strategy for PHT to induce cancer cell death.
showed moderate nucleocytoplasmatic relation, pleomorphic nuclei (tetra-lobed nuclei) and basophilic cytoplasm without granules (Figure 1); (2) negative serology for Chagas disease, human immunodeiciency virus (HIV) 1 and 2, syphilis and hepatitis; (3) tests for antinuclear factor, Ro/SS-A and La/SS-B autoantibodies, anti-deoxyribonucleic acid, rheumatoid factor and antistreptolysin O were also non reactive; (4) negative C reactive protein (CRP); (5) negative direct Coombs; (6) negative lupus anticoagulant. On the same day of collection, other exams were required (hemogram, myelogram and immunophenotyping by low cytometry) in order to exclude the diagnostic possibility of chronic lymphocytic leukemia/ small lymphocytic lymphoma.
Giant Cell Reparative Granuloma (GCRG) of phalanx is uncommon. It is a benign osteolytic lesion but can be locally aggressive. GCRG has certain radiology and histological features that are similar to other giant cell lesions of the bone. We present a case report of a young patient with giant cell reparative granuloma of proximal phalanx of left third toe. The bone lesion was successfully treated surgically. Key Words: