Clinical features predict responsiveness to imatinib in platelet-derived growth factor receptor-alpha-negative hypereosinophilic syndrome.

BACKGROUND: With the exception of the presence of the FIP1L1-PDGFRA fusion gene, little is known about predictors of imatinib response in clinically-defined hypereosinophilic syndrome (HES). METHODS: Subjects with FIP1L1-PDGFRA-myeloid neoplasm (FP; n =12), PDGFRA-negative HES with ≥4 criteria suggestive of a myeloid neoplasm (MHES; n =10), or steroid-refractory PDGFRA-negative HES with <4 myeloid criteria (SR; n = 5) were enrolled in a prospective study of imatinib therapy (NCT00044304: registered at clinicaltrials.gov). The primary outcome was an eosinophil count <1.5 × 109/L at one month and improvement of clinical symptoms. Clinical, molecular, and bone marrow responses to imatinib were assessed. A retrospective cohort of 18 subjects with clinically-defined HES who received imatinib (300-400 mg daily ≥ 1 month) were classified according to the criteria used in the prospective study. RESULTS: Overall, imatinib response rates were 100% in the FP group (n = 16), 54% in the MHES group (n = 13) and 0% in the SR group (n = 16). The presence of ≥ 4 myeloid features was the sole predictor of response. After ≥ 18 months in complete remission, imatinib was tapered and discontinued in 8 FP and 1 MHES subjects. Seven subjects (6 FP, 1 MHES) remain in remission off therapy for a median of 29 months (range 14-36). CONCLUSIONS: Clinical features of MHES predict imatinib response in PDGFRA-negative HES.

The cellular immune system in myelomagenesis: NK cells and T cells in the development of myeloma [corrected] and their uses in immunotherapies.

As vast strides are being made in the management and treatment of multiple myeloma (MM), recent interests are increasingly focusing on understanding the development of the disease. The knowledge that MM develops exclusively from a protracted phase of monoclonal gammopathy of undetermined significance provides an opportunity to study tumor evolution in this process. Although the immune system has been implicated in the development of MM, the scientific literature on the role and status of various immune components in this process is broad and sometimes contradictory. Accordingly, we present a review of cellular immune subsets in myelomagenesis. We summarize the current literature on the quantitative and functional profiles of natural killer cells and T-cells, including conventional T-cells, natural killer T-cells, γδ T-cells and regulatory T-cells, in myelomagenesis. Our goal is to provide an overview of the status and function of these immune cells in both the peripheral blood and the bone marrow during myelomagenesis. This provides a better understanding of the nature of the immune system in tumor evolution, the knowledge of which is especially significant considering that immunotherapies are increasingly being explored in the treatment of both MM and its precursor conditions.

Ibrutinib-induced lymphocytosis in patients with chronic lymphocytic leukemia: correlative analyses from a phase II study.

Ibrutinib and other targeted inhibitors of B-cell receptor signaling achieve impressive clinical results for patients with chronic lymphocytic leukemia (CLL). A treatment-induced rise in absolute lymphocyte count (ALC) has emerged as a class effect of kinase inhibitors in CLL and warrants further investigation. Here we report correlative studies in 64 patients with CLL treated with ibrutinib. We quantified tumor burden in blood, lymph nodes (LNs), spleen and bone marrow, assessed phenotypic changes of circulating cells and measured whole-blood viscosity. With just one dose of ibrutinib, the average increase in ALC was 66%, and in>40% of patients the ALC peaked within 24 h of initiating treatment. Circulating CLL cells on day 2 showed increased Ki67 and CD38 expression, indicating an efflux of tumor cells from the tissue compartments into the blood. The kinetics and degree of the treatment-induced lymphocytosis was highly variable; interestingly, in patients with a high baseline ALC the relative increase was mild and resolution rapid. After two cycles of treatment the disease burden in the LN, bone marrow and spleen decreased irrespective of the relative change in ALC. Whole-blood viscosity was dependent on both ALC and hemoglobin. No adverse events were attributed to the lymphocytosis.

Mutation frequencies of GNAQ, GNA11, BAP1, SF3B1, EIF1AX and TERT in uveal melanoma: detection of an activating mutation in the TERT gene promoter in a single case of uveal melanoma.

BACKGROUND: Uveal melanoma is the most frequent primary tumour of the eye. It is molecularly clearly distinct from cutaneous melanoma and shows a different pattern of driver mutations. The influence of sunlight ultraviolet (UV) exposure on the aetiology of uveal melanoma is a matter of debate. The recent identification of driver mutations in the promoter of the telomerase reverse transcriptase (TERT) gene with UV-induced cytidine-to-thymidine transitions in cutaneous melanoma prompted us to investigate whether these mutations also occur in uveal melanoma. METHODS: We analysed 50 cases of uveal melanoma obtained from enucleation surgery for mutations in the genes GNAQ, GNA11, BAP1, SF3B1, EIFAX1 and TERT, measured gene expression using microarrays and analysed gene copy numbers by SNP arrays. RESULTS: We detected a TERT mutation in only one case of a 57-year-old white male patient with clinical and histopathological features typical for uveal melanoma. The tumour showed mutations in GNA11 and EIF1AX that are typical for uveal melanoma and absent from cutaneous melanoma. No mutations were detected in GNAQ, BAP1 and SF3B1 that are frequently mutated in uveal melanoma. Both copies of chromosome 3 were retained. Several tumours among which the one carrying the TERT promoter mutation showed elevated TERT expression. Consistent with previous reports, GNAQ is inversely associated with chromosome 3 monosomy and metastasis. BAP1 mutations are significantly associated with chromosome 3 monosomy but not with relapse. CONCLUSION: These data indicate that TERT mutations are rare in uveal melanoma. No conclusion can be drawn on their potential influence on tumour progression.

Expression of receptor activator of nuclear factor-kappa B ligand in leukocytes during acute kidney rejection after transplantation in rats.

BACKGROUND: Acute cellular rejection of the transplanted kidney is an important cause of impaired graft function. One of the basic characteristics of acute cellular rejection according to the latest Banff classification of renal allograft pathology is the presence of a large number of T lymphocytes in the allografted tissue. Osteoprotegerin, receptor activator of nuclear factor-kappa B (RANK) and RANK ligand (RANKL), three relatively novel members of the tumor necrosis factor superfamily, have crucial roles not only in physiologic and pathologic bone metabolism but also in immunologic processes. The aim of our study was to determine the expression of RANKL and RANK by T lymphocytes and macrophages in acute cellular kidney allograft rejection in rats. METHODS: The study included 15 male Wistar rats of 3 months old and 250-300 g as recipients and 15 male DA rats donors of 3 months old; and weight 250-300 g. When animals were sacrificed at 3 weeks to extract the transplanted kidney for pathohistologic analysis and immunoflorescence. all samples showed acute cellular rejection. Kidney sections were examined by dual-labeled immunofluorescence to detect CD4, CD8, or CD68 (red) and RANK or RANKL (green) with coexpressing cells as orange. RESULTS: RANKL-positive expression colocalized with CD4(+) and CD8(+) T lymphocytes in acutely rejected kidney tissue. There was no association between CD4(+) and CD8(+) T cells with RANK expression, which was evident by infiltrating CD68-positive macrophages in the kidney tissue interstitium. CONCLUSION: RANK and RANKL were expressed by T lymphocytes and macrophages in acute cellular kidney rejection after transplantation in rats.

Targeting the KIT activating switch control pocket: a novel mechanism to inhibit neoplastic mast cell proliferation and mast cell activation.

Activating mutations in the receptor tyrosine kinase KIT, most notably KIT D816V, are commonly observed in patients with systemic mastocytosis. Thus, inhibition of KIT has been a major focus for treatment of this disorder. Here we investigated a novel approach to such inhibition. Utilizing rational drug design, we targeted the switch pocket (SP) of KIT, which regulates its catalytic conformation. Two SP inhibitors thus identified, DP-2976 and DP-4851, were examined for effects on neoplastic mast cell proliferation and mast cell activation. Autophosphorylation of both wild-type and, where also examined, KIT D816V activation was blocked by these compounds in transfected 293T cells, HMC 1.1 and 1.2 human mast cell lines, and in CD34(+)-derived human mast cells activated by stem cell factor (SCF). Both inhibitors induced apoptosis in the neoplastic mast cell lines and reduced survival of primary bone marrow mast cells from patients with mastocytosis. Moreover, the SP inhibitors more selectively blocked SCF potentiation of FcɛRI-mediated degranulation. Overall, SP inhibitors represent an innovative mechanism of KIT inhibition whose dual suppression of KIT D816V neoplastic mast cell proliferation and SCF-enhanced mast cell activation may provide significant therapeutic benefits.

Today Balkan endemic nephropathy is a disease of the elderly with a good prognosis.

BACKGROUND: The outcome for Balkan endemic nephropathy (BEN) patients diagnosed in 1992 was analyzed in 2006 with the aim of detecting factors associated with disease progression and patient outcome. METHODS: In 1992 BEN was detected in 119 patients (53 males, 56.9 +/- 13.8 years) from the village of Sopic. Changes in creatinine clearance as well as outcome (death or onset of regular hemodialysis) were analyzed retrospectively in 2006. RESULTS: During the 14-year period 47 patients deceased (5 on hemodialysis) at the mean age of 72.2 +/- 8.2 years, while no data were available for 13 cases. Out of 59 remaining patients 3 were on hemodialysis in 2006 and 56 participated in the control examination. Of these 12 had creatinine clearance at least 50% lower than in 1992 and 44 had unchanged creatinine clearance. Logistic regression revealed age and proteinuria, but linear regression only age as significant prognostic factors for changes in creatinine clearance. The all-cause mortality rate varied between 1.1 and 5.3% per year and was similar to the mortality rate of the general population. The main cause of death was cardiovascular disease (40.5%) followed by malignant diseases (17%), most frequently (11%) due to upper urothelial tumors. Urine protein and age were found to be a significant independent predictor of all-cause mortality. CONCLUSION: In the village of Sopic BEN was commonly detected in patients in their fifties, progressed slowly, but most patients died from other causes at old age before end-stage renal disease occurred. Kidney function remained stable over the decade in three quarters of the surviving patients. Age and proteinuria were found to be prognostic factors for both disease progression and patient mortality.

Efficacy of hyperphosphatemia control in the progression of chronic renal failure and the prevalence of cardiovascular calcification.

BACKGROUND AND AIMS: Chronic kidney disease mineral- and bone disorder (CKD-MBD) has been studied more often in dialysis than in predialysis CKD patients. The association between efficacy of hyperphosphatemia control and chronic renal failure (CRF) progression, prevalence of bone disease and cardiovascular calcification was the objective of the present investigation. MATERIAL AND METHODS: 42 patients with CKD in Stage 5, regularly monitored for 5 years, were divided into Group 1 of 20 patients with normal serum phosphate (sPO4) levels and Group 2 of 22 patients with hyperphosphatemia registered at the majority of checks. Serum urea, creatinine, calcium (sCa) and sPO4 levels were regularly determined in the retrospective 5-year period. At the end of this period iPTH, bone alkaline phosphatase-BAP and inflammation markers (CRP, fetuin-A) were measured, valvular and arterial calcifications were detected by B mode echocardiogram and soft-tissue native radiograms of the pelvis and the wrist. RESULTS: Progression of CRF (1/sCr over time) was faster in Group 2 than in Group 1 (b = -0.0577 vs. -0.0288, p = 0.003) during the study period. Average BAP and iPTH values were similar in both groups and 23/42 patients had PTH > 300 pg/ml. Arterial and valvular calcifications were found in 5/23 patients from Group 1 and 14/22 patients from Group 2 (p = 0.011). Linear regression analysis revealed sPO4 as a predictor for total calcification number, inflammatory diseases as a predictor for valvular calcifications, while sPO4 and iPTH were predictors for arterial calcifications. CONCLUSIONS: More than half the patients with Stage 5 CKD not yet on dialysis exhibited elevated PTH. Faster CRF progression and frequent arterial and valvular calcifications were seen in patients with poor phosphate control and sPO4 was selected as an independent predictor of total calcification score.

[Massive GIST of the stomach--case report].

Gastrointestinal stromal tumors (GIST) are rare mesenchimal neoplasmas of the gastrointestinal tract. The diagnosis of this tumors are oftenly very difficult. Patients with this tumor are ussualy addmited to the hospital cause of the gastrointestinal bleeding, abdominal pain, abdominal distension, disphagia, obstructive jaundice and bowel obsstruction. In this case report, we present a 86 year old patient with massive GIST of the stomach which was not preoperatively diagnosed.

Stereotypical physiological properties emerge during early neuronal and glial lineage development in the embryonic rat neocortex.

Surface immunolabeling was used together with membrane potential and/or Ca(2+) indicator dyes to characterize physiological properties emerging among precursors, neuroglial progenitors and differentiating neurons during neurogenesis of embryonic rat neocortex. Cells were immunoidentified with tetanus toxin (TnTx), which binds to gangliosides expressed by neurons, and anti-A2B5, which reacts with gangliosides expressed by neuroglial progenitors. Microdissection of the neocortex into ventricular/subventricular zone (VZ/SVZ) and cortical plate/subplate (CP/SP) regions further resolved the TnTx/A2B5-immunoidentified cells into pre- and post-migratory subpopulations. Quantitative immunocytochemistry revealed mainly proliferative (BrdU(+)) and immature (nestin(+)) elements among TnTx(-)A2B5(-) precursors and TnTx(-)A2B5(+) progenitors in the VZ/SVZ, and the appearance of neuron-specific antigens among post-mitotic TnTx(+) subpopulations of the CP/SP. Flow cytometry of acutely prepared cells in suspension and dual-imaging of cells in culture revealed that ionotropic amino acid receptors and metabotropic acetylcholine receptors closely paralleled the emergence of voltage-dependent Na(+) and Ca(2+) channels and Na(+)-Ca(2+) exchange activity among TnTx(+) neuronal progenitors migrating from VZ/SVZ to CP/SP. During this period, TnTx(-)A2B5(-) precursors and TnTx(-)A2B5(+) neuroglial progenitors from VZ/SVZ predominantly exhibited Ca(2+) responses to ATP. Thus, stereotypical and contrasting physiologies emerge among embryonic cortical cells in vivo as they initially progress from proliferating precursors and progenitors along neuronal and glial cell lineages.

Variations of the great arteries in the carotid triangle.

The variations of the common carotid artery, as well as of the external and internal carotid arteries, are described. During anatomic dissection on adult cadavers, we investigated the variability of appearance of 40 carotid arterial systems. Special consideration was given to the topographic relations such as the level of the bifurcation of the common carotid artery, the relationship between the external and internal carotid arteries, and the origin of the great collateral branches. Special attention was paid to the origin of the superior thyroid artery. In this article the practical importance of these variations is stressed.

Increased TUNEL staining in brains of autoimmune Fas-deficient mice.

Profound changes in brain morphology and behavior coincide with the spontaneous development of systemic autoimmune/inflammatory disease in Fas-deficient MRL-lpr mice. The dendrites atrophy, the density of hippocampal and cortical neurons decreases, and an anxious/depressive-like behavior emerges while lymphoid cells infiltrate into the choroid plexus of MRL-lpr mice. We hypothesized that the inherited lack of the Fas-dependent anti-inflammatory mechanism would lead to unsuppressed immune activity, characterized by reduced apoptosis in the MRL-lpr brain. Using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeled (TUNEL) method as an indicator of apoptosis, a surprisingly high incidence of TUNEL-positive cells was observed in the hippocampus, choroid plexus and periventricular regions of MRL-lpr mice, 5-10-fold higher than that found in the MRL +/+ control brain. Immunostaining with anti-CD3, CD4 and CD8 monoclonal antibodies showed limited overlap between CD-positive and TUNEL-positive cells, suggesting that the dying cells are for the most part (approximately 70%) not T-lymphocytes. Although further characterization of the phenotype of the dying cells and the mechanism of cell death are required, the present results suggest the involvement of a Fas-independent apoptotic process in neurodegeneration induced by systemic autoimmune disease.

Buoyant density gradient fractionation and flow cytometric analysis of embryonic rat cortical neurons and progenitor cells.

We have used the property of natural cell buoyant density to selectively fractionate embryonic rat neocortical cells into 20 subpopulations ranging in phenotype from proliferatively active progenitors to terminally postmitotic neurons. Immunocytochemical and cell cycle analysis of the cellular fractions with flow cytometry revealed an inverse relationship between cell buoyant density and neuronal differentiation. The most buoyant fractions contained predominantly terminally postmitotic, tubulin betaIII-positive, tetanus toxin-positive, and nestin-negative differentiating neurons, while immature, bromodeoxyuridine-positive and nestin-positive proliferating cells were more prevalent in less buoyant fractions. Double loading of isolated cells with voltage- and Ca2+-sensitive fluorescent indicator dyes followed by simultaneous recordings of membrane potential and cytoplasmic [Ca2+] ([Ca2+]c]) using flow cytometry revealed that >50% of the least buoyant cells produced functional responses to veratridine, a Na+ channel agonist, and muscimol, a GABA(A) receptor agonist, but <10% responded to kainic acid, an agonist of a subset of glutamate receptors. As cells became more buoyant the percentage of cells that depolarized and produced a rise in [Ca2+]c to each ligand increased, particularly in response to kainic acid. Short-term culture of select fractions revealed a marked enrichment for cells with morphologies and epitopes characteristic of neuronal and progenitor cell subpopulations. The results show that embryonic cortical cells exhibit a range of naturally occurring buoyant densities that can be used to expeditiously fractionate cortical cells according to their pre- or postmitotic status, thus providing ready access for cellular and molecular studies of proliferation and differentiation.

[Urinary system tumors in patients with endemic nephropathy on chronic hemodialysis]. / Tumori urinarnog sistema kod bolesnika s endemskom nefropatijom i na hronicnoj hemodijalizi.

We studied 161 patients who started dialysis from January 1st 1979 to December 31st 1993. Out of 79 males, 26 (32.91%) had the diagnosis of urothelial tumors, and out of these 22 patients (84.61%) had tumors of the upper urothelium. Twenty one (80.76%) of the patients were operated. The predialysis period starting with tumor diagnosing was 61.92 months averaging. The average survival of these hemodialysed patients was 46.07 months. Out of 82 females, 22 (26.82) had tumors of the urinary tract, and out of these 21 patients (95.45%) had tumors of the upper urothelium. Eighteen (81.81%) of them were operated. The predialysis period starting with tumor diagnosing was 84 months, on the average. The average survival was 37.39 months. The results confirm high frequency of tumors of the upper urothelium in patients with endemic nephropathy.