bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic aberrations common in gastric high-grade large B-cell lymphoma
Authors: Starostik Petr , Greiner Axel , Schultz Anja , Zettl Andreas , Peters Katharina , Rosenwald Andreas , Kolve Mariele , Müller-Hermelink Hans Konrad
Published: October 2019

Genetic aberrations associated with the development of extranodal high-grade large B-cell lymphoma originating in the stomach have not been fully identified yet. We analyzed 31 such lymphomas using 73 microsatellite markers for allelic imbalance and microsatellite instability. The highest frequency (42%) of loss of heterozygosity (LOH) was found on the long arm of chromosome 6. We identified 2 LOH hot spots on 6q21-22.1 and 6q23.3-25, flanked by markers D6S246-D6S261 and D6S310-D6S441, respectively, containing putative tumor suppressor genes (TSGs). These 6q aberrations were found to be the sole allelic imbalance in 1 patient only; they were mostly accompanied by additional abnormalities. Several known TSGs, namely, the APC, p15/p16, p53, and DCC genes, were found to suffer frequent LOH during lymphomagenesis. LOH was also detected in regions containing putative TSGs on 7q and 13q14. Frequent amplification of genomic material was found in the 2p, 3q27 at the BCL-6 gene locus, 6p, 7q, 11q23-24 at the MLL gene locus, and 18q regions. Analysis of the pattern of occurrence of these aberrations revealed an association of the amplification of the MLL gene region with LOH at the p53 locus (P = .02). Only low frequency of microsatellite instability (MSI) was detected in these lymphomas and MSI incidence increased with age (P = .01). Karyotypic instability thus plays the main role in the development of gastric high-grade large B-cell lymphoma. Common genetic aberrations responsible for lymphomagenesis are deletions of 6q, loss of p53, and amplification of the 3q27 and the MLL gene regions.
Actin cytoskeletal function is spared, but apoptosis is increased, in WAS patient hematopoietic cells
Authors: Rengan Ramesh , Ochs Hans D. , Sweet Leonard I. , Keil Michael L. , Gunning William T. , Lachant Neil A. , Boxer Laurence A. , Omann Geneva M.
Published: October 2019

Mutations in the Wiskott-Aldrich syndrome protein (WASP) have been hypothesized to cause defective actin cytoskeletal function. This resultant dysfunction of the actin cytoskeleton has been implicated in the pathogenesis of Wiskott-Aldrich syndrome (WAS). In contrast, it was found that stimulated actin polymerization is kinetically normal in the hematopoietic lineages affected in WAS. It was also found that the actin cytoskeleton in WAS platelets is capable of producing the hallmark cytoarchitectural features associated with activation. Further analysis revealed accelerated cell death in WAS lymphocytes as evidenced by increased caspase-3 activity. This increased activity resulted in accelerated apoptosis of these cells. CD95 expression was also increased in these cells, suggesting an up-regulation in the FAS pathway in WAS lymphocytes. Additionally, inhibition of actin polymerization in lymphocytes using cytochalasin B did not accelerate apoptosis in these cells. This suggests that the accelerated apoptosis observed in WAS lymphocytes was not secondary to an underlying defect in actin polymerization caused by mutation of the WAS gene. These data indicate that WASP does not play a universal role in signaling actin polymerization, but does play a role in delaying cell death. Therefore, the principal consequence of mutations in theWAS gene is to accelerate lymphocyte apoptosis, potentially through up-regulation of the FAS-mediated cell death pathway. This accelerated apoptosis may ultimately give rise to the clinical manifestations observed in WAS.
Stimulation of cytotoxic T cells against idiotype immunoglobulin of malignant lymphoma with protein-pulsed or idiotype-transduced dendritic cells
Authors: Osterroth Frank , Garbe Annette , Fisch Paul , Veelken Hendrik
Published: October 2019

Because of their hypervariable regions and somatic mutations, the antigen receptor molecules of lymphomas (idiotypes) are tumor-specific antigens and attractive targets for antilymphoma immunotherapy. For the optimal induction of human idiotype-specific cytotoxic T cells (CTL), idiotype was presented to CD8+ peripheral blood mononuclear cells by monocyte-derived autologous dendritic cells (DC) after the endocytosis of idiotype protein or by idiotype-expressing DC. Recombinant idiotype was obtained as a functionally folded Fab fragment by periplasmic expression in Escherichia coli. Idiotype-expressing DC were generated by transduction with recombinant Semliki forest virus vectors encompassing heavy- or light-chain idiotype genes. Autologous lymphoblastoid cell lines stably transfected with Epstein-Barr virus-based idiotype expression vectors were used as target cells to detect idiotype-specific lysis. CTL stimulated with idiotype-loaded DC showed strong specific, CD8-mediated, and major histocompatibility complex (MHC) class I-restricted cytotoxicity against autologous heavy- and light-chain idiotype. In contrast, stimulation with idiotype-transduced DC resulted in only moderate natural killer cell activity. These data confirm the existence of idiotype-specific CTL in patients with lymphoma, define a “good manufacturing practice”-compatible protocol for the generation of these cells without the requirement of viable lymphoma cells, and favor the processing of exogenous antigen over DC transduction for the induction of MHC I-restricted CTL against idiotypes with unknown antigenicity.
A soluble form of human Delta-like-1 inhibits differentiation of hematopoietic progenitor cells
Authors: Han Wei , Ye Qian , Moore Malcolm A. S.
Published: October 2019

Two Notch ligand families, Delta and Serrate/Jagged, have been identified in vertebrates. Members of the Jagged family have been shown to affect in vitro hematopoiesis. To determine whether members of the Delta family might play a similar role in hematopoiesis, we examined the expression of mouse Delta-like-1 (mDll1). mDll1 protein was detected in whole marrow and in a marrow stromal cell line MS-5. At the RNA level, both mDll1 and Notch1 were seen in marrow precursor, differentiated hematopoietic, marrow stromal, and MS-5 cells. We isolated a cDNA encoding the human homologue of mDll1, designated human Delta-like-1 (hDll1). A soluble form of hDll1, hDll1NDSL, containing the DSL domain and the N-terminal sequences, was expressed and purified from bacteria as a glutathione S-transferase (GST) fusion protein. We observed that hDll1NDSL delayed the acquisition of differentiation markers by murine hematopoietic progenitor cells (Lin−) cultured in vitro with cytokines. In addition, it promoted greater expansion (more than 3 times) of the primitive hematopoietic precursor cell population, measured in high-proliferative potential colony assay and day 12 colony-forming unit spleen (CFU-S) assay, than GST controls. We also observed that the percentage of apoptotic cells decreased and that the number of cells in the S-phase of the cell cycle increased in the cultures of Lin−cells with hDll1NDSL. The effects of hDll1NDSL were blocked by antibody against the mouse counterpart of hDll1NDSL, mDll1NDSL. These observations demonstrate that hDll1 plays a role in mediating cell fate decisions during hematopoiesis.
A monoclonal antibody specific to the granulocyte-derived elastase-fragment D species of human fibrinogen and fibrin: its application to the measurement of granulocyte-derived elastase digests in plasma
Authors: Kohno Isao , Inuzuka Kimiko , Itoh Yumiko , Nakahara Kunihiko , Eguchi Yutaka , Sugo Teruko , Soe Gilbu , Sakata Yoichi , Murayama Hideki , Matsuda Michio
Published: October 2019

When granulocytes are stimulated under certain clinical conditions, elastase is released therefrom and digests fibrin(ogen) independently of the plasmin system, which may also be mobilized simultaneously. Thus, discrimination of these 2 systems becomes urgent for the diagnosis and treatment of the underlying diseases. Using as immunogen a 97-kd granulocyte-elastase digest of human fibrinogen, we raised an antibody IF-123 that specifically recognizes elastase digests of human fibrin(ogen). The 97-kd elastase fragment resembles plasmic fragment D1, and the epitope of this antibody is located on the A (196-204) residue segment. This segment appears to be masked in fibrin(ogen) but exposed when the A Leu 204-Ile 205 peptide bond is cleaved by elastase. Cathepsin G concomitantly released from granulocytes failed to expose the epitope. By an enzyme immunoassay using IF-123 as the capture antibody, the elastase digests of fibrin(ogen) can be measured in plasma samples without interference by abundantly coexisting fibrinogen. Indeed, we found that the elastase digests were mostly elevated in patients with inflammation or malignant tumors, but remained in a normal range in patients with a benign gastrointestinal tract disease such as duodenal ulcer and polyps in the gallbladder or the colon. Like the plasmic D-dimer, the elastase digests predominantly consisted of the DD/E complex and DD/E-containing high-molecular weight derivatives apparently corresponding to the phase-3 plasmic digests of cross-linked fibrin.
Lepirudin blunts endotoxin-induced coagulation activation
Authors: Pernerstorfer T. , Hollenstein U. , Hansen J.-B. , Stohlawetz P. , Eichler H.-G. , Handler S. , Speiser W. , Jilma B.
Published: October 2019

During sepsis, lipopolysaccharide (LPS) triggers the development of disseminated intravascular coagulation (DIC) via the tissue factor-dependent pathway of coagulation resulting in massive thrombin generation and fibrin polymerization. Recently, animal studies demonstrated that hirudin reduced fibrin deposition in liver and kidney and decreased mortality in LPS-induced DIC. Accordingly, the effects of recombinant hirudin (lepirudin) was compared with those caused by placebo on LPS-induced coagulation in humans. Twenty-four healthy male subjects participated in this randomized, double-blind, placebo-controlled, parallel group study. Volunteers received 2 ng/kg LPS intravenously, followed by a bolus-primed continuous infusion of placebo or lepirudin (Refludan, bolus: 0.1 mg/kg, infusion: 0.1 mg/kg/h for 5 hours) to achieve a 2-fold prolongation of the activated partial thromboplastin time (aPTT). LPS infusion enhanced thrombin activity as evidenced by a 20-fold increase of thrombin-antithrombin complexes (TAT), a 6-fold increase of polymerized soluble fibrin, termed thrombus precursor protein (TpP), and a 4-fold increase in D-dimer. In the lepirudin group, TAT increased only 5-fold, TpP increased by only 50%, and D-dimer only slightly exceeded baseline values (P < .01 versus placebo). Concomitantly, lepirudin also blunted thrombin generation evidenced by an attenuated rise in prothrombin fragment levels (F1 + 2,P < .01 versus placebo) and blunted the expression of tissue factor on circulating monocytes. This experimental model proved the anticoagulatory potency of lepirudin in LPS-induced coagulation activation. Results from this trial provide a rationale for a randomized clinical trial on the efficacy of lepirudin in DIC.
FGFR1 is fused to the centrosome-associated proteinCEP110 in the 8p12 stem cell myeloproliferative disorder with t(8;9)(p12;q33)
Authors: Guasch Géraldine , Mack Gary J. , Popovici Cornel , Dastugue Nicole , Birnbaum Daniel , Rattner Jérome B. , Pébusque Marie-Josèphe
Published: October 2019

The hallmark of the 8p12 stem cell myeloproliferative disorder (MPD) is the disruption of the FGFR1 gene, which encodes a tyrosine kinase receptor for members of the fibroblast growth factor family.FGFR1 can be fused to at least 3 partner genes at chromosomal regions 6q27, 9q33, or 13q12. We report here the cloning of the t(8;9)(p12;q33) and the detection of a novel fusion betweenFGFR1 and the CEP110 gene, which codes for a novel centrosome-associated protein with a unique cell-cycle distribution. CEP110 is widely expressed at various levels in different tissues and is predicted to encode a 994-amino acid coiled-coil protein with 4 consensus leucine zippers [L-X(6)-L-X(6)-L-X(6)-L]. Both reciprocal fusion transcripts are expressed in the patient's cells. The CEP110-FGFR1 fusion protein encodes an aberrant tyrosine kinase of circa 150-kd, which retains most of CEP110 with the leucine zipper motifs and the catalytic domain of FGFR1. Transient expression studies show that the CEP110-FGFR1 protein has a constitutive kinase activity and is located within the cell cytoplasm.
Cell adhesion receptors in lymphoma dissemination
Authors: Drillenburg Paul , Pals Steven T.
Published: October 2019

Regulated lymphocyte trafficking is essential for the control and integration of systemic immune responses. This homing process disperses the immunologic repertoire, guides lymphocyte subsets to the specialized microenvironments that control their differentiation and survival, and targets immune effector cells to sites of antigenic insult. This review discusses data indicating that the adhesion receptors regulating the trafficking of normal lymphocytes are also expressed and functionally active in their malignant counterparts, the non-Hodgkin lymphomas. These “homing receptors” appear to mediate the highly tissue-specific dissemination of specific lymphoma subtypes, such as lymphomas of the mucosa-associated lymphoid tissues and lymphomas of the skin. Furthermore, as a result of their capability to enhance lymphoma dissemination and to transduce signals into the cell, promoting cell growth and survival, adhesion receptors may contribute to lymphoma aggressiveness. Taken together, the data offer a framework for understanding the dissemination routes of non-Hodgkin lymphomas and suggest that adhesion receptors, specifically those of the CD44 family, may present useful tools to predict prognosis in patients with lymphomas.
Genetic analysis, phenotypic diagnosis, and risk of venous thrombosis in families with inherited deficiencies of protein S
Authors: Makris Michael , Leach Michael , Beauchamp Nick J. , Daly Martina E. , Cooper Peter C. , Hampton Kingsley K. , Bayliss Pauline , Peake Ian R. , Miller George J. , Preston F. Eric
Published: October 2019

Protein S deficiency is a recognized risk factor for venous thrombosis. Of all the inherited thrombophilic conditions, it remains the most difficult to diagnose because of phenotypic variability, which can lead to inconclusive results. We have overcome this problem by studying a cohort of patients from a single center where the diagnosis was confirmed at the genetic level. Twenty-eight index patients with protein S deficiency and a PROS1 gene defect were studied, together with 109 first-degree relatives. To avoid selection bias, we confined analysis of total and free protein S levels and thrombotic risk to the patients' relatives. In this group of relatives, a low free protein S level was the most reliable predictor of a PROS1gene defect (sensitivity 97.7%, specificity 100%). First-degree relatives with a PROS1 gene defect had a 5.0-fold higher risk of thrombosis (95% confidence interval, 1.5-16.8) than those with a normal PROS1 gene and no other recognized thrombophilic defect. Although pregnancy/puerperium and immobility/trauma were important precipitating factors for thrombosis, almost half of the events were spontaneous. Relatives with splice-site or major structural defects in the PROS1 gene were more likely to have had a thrombotic event and had significantly lower total and free protein S levels than those relatives having missense mutations. We conclude that persons withPROS1 gene defects and protein S deficiency are at increased risk of thrombosis and that free protein S estimation offers the most reliable way of diagnosing the deficiency.
Identification of a family of alternatively spliced mRNA species of angiopoietin-1
Authors: Huang Yao-Qi , Li Jian-Jun , Karpatkin Simon
Published: October 2019

Angiopoietin-1 (Ang-1) is required for developing vessels, and its absence leads to defects in vessel remodeling. Ang-1 has been identified as the ligand for the tyrosine kinase receptor Tie-2, which is expressed specifically on endothelial cells and early hematopoietic cells. In studying the role of Tie-2 and Ang-1 in megakaryocytopoiesis, 3 alternatively spliced species of Ang-1 mRNA (Ang-1.3 kb, Ang-0.9 kb, and Ang-0.7 kb) were identified in addition to the full-length Ang-1 (Ang-1.5 kb), in the megakaryocyte cell line CHRF by reverse transcription–polymerase chain reaction (RT-PCR), and then cloned and sequenced. The expression of 3 alternatively spliced isoforms of Ang-1 was confirmed by RT-PCR using specific primer pairs derived from junction sites and the 3′ end of Ang-1 cDNA, and it was further demonstrated by nuclease protection assay, Northern blotting, and immunoblotting in CHRF cells. Expression of the Ang-1.3 kb isoform was also detected in human primary fibroblast cell line FS4, breast cancer cell line MDAMB-468, and CD34+CD41+ cells of fetal liver and platelets. The function of the 1.5-kb, 1.3-kb, and 0.9-kb isoforms was examined. Recombinant proteins Ang-1.5 and 0.9 kb bind strongly to the recombinant Tie-2 receptor (Tie-2-Fc), whereas the 1.3-kb isoform does not. The Ang-1.3 kb isoform binds to the 1.5-kb isoform. Ang-1.5 kb, but not the 1.3-kb and 0.9-kb isoforms, induces tyrosine phosphorylation of Tie-2 in human umbilical vein endothelial cells. These data suggest that isoforms 1.3 kb and 0.9 kb could serve as dominant negative molecules for the full-length Ang-1. The possible involvement of the newly identified Ang-1 isoforms in angiogenesis and in growth and differentiation of hematopoietic progenitor cells provides a greater complexity to these processes.
A critical role for PI 3-kinase in cytokine-induced Fcα-receptor activation
Authors: Bracke Madelon , Nijhuis Evert , Lammers Jan-Willem J. , Coffer Paul J. , Koenderman Leo
Published: October 2019

Fc-receptors, such as FcR and FcγRII, play an important role in leukocyte activation, and rapid modulation of ligand binding (“activation”) is critical for receptor regulation. We have previously demonstrated that ligand binding to Fc-receptors on human eosinophils is dependent on cytokine stimulation. Utilization of pharmacological inhibitors provided evidence that the phenomenon of interleukin (IL)-5 induced immunoglobulin A (IgA) binding to human eosinophils requires activation of phosphatidylinositol 3-kinase (PI3K). However, eosinophils are refractory to manipulation by molecular techniques such as DNA transfection or viral infection. Here we utilize an IL-3 dependent pre-B cell line to investigate the molecular mechanism of cytokine-mediated ligand binding to FcR. In this system, IgA binding is dependent on IL-3, similarly to the requirement for IL-5 of eosinophils. We show that IL-3-mediated activation of FcR (CD89) requires the activation of PI3K, independent of p21ras activation. Co-expression of dominant negative (▵p85) and active (p110_K227E) forms of PI3K demonstrate that the affinity switch regulating FcR activation requires PI3K. Moreover, overexpression of PI3K is both necessary and sufficient for activation of FcR. Furthermore, we show that IL-3/IL-5/GM-CSF induced inside-out signaling pathways activating FcR require the involvement of protein kinase C downstream of PI3K. Finally, we show that these inside-out signaling pathways responsible for Fc-receptor modulation require CD89, independent of its association with the FcRγ chain.
STAT5 activation contributes to growth and viability in Bcr/Abl-transformed cells
Authors: Sillaber Christian , Gesbert Franck , Frank David A. , Sattler Martin , Griffin James D.
Published: October 2019

The transcription factor STAT5 is constitutively tyrosine phosphorylated and activated after transformation of hematopoietic cells by p210Bcr/Abl. A truncated form of STAT5B (▵STAT5; aa, 1-683) that lacks tyrosine 699 and the transcriptional activation domain was introduced into Ba/F3p210 cells under the control of a tetracycline-inducible promoter. Treatment of these cells with doxycycline, a tetracycline analogue, induced expression of ▵STAT5 and inhibited STAT5-dependent transcription. ▵STAT5 coprecipitated with STAT5 and decreased Bcr/Abl-dependent tyrosine phosphorylation of endogenous STAT5. Induction of ▵STAT5 inhibited growth of Ba/F3p210 cells (26%-52% of control levels at 4 days) but did not cause cell-cycle arrest. ▵STAT5 reduced viability of Ba/F3p210 cells and increased sensitivity of the cells to the cytotoxic drugs hydroxyurea and cytarabine. These results indicate that high-level expression of ▵STAT5, as achieved here by using a tetracycline-inducible promoter, inhibits STAT5 activity, reduces the growth rate of Ba/F3p210 cells by inhibiting viability, and results in increased sensitivity to chemotherapeutic drugs. It is therefore likely that STAT5 activation plays a role in the transformation of hematopoietic cell lines by p210Bcr/Abl.
Granulocyte colony-stimulating factor receptor mutations in severe congenital neutropenia transforming to acute myelogenous leukemia confer resistance to apoptosis and enhance cell survival
Authors: Hunter Melissa G. , Avalos Belinda R.
Published: October 2019

Patients with severe congenital neutropenia (SCN) are at increased risk for the development of acute myelogenous leukemia (AML). In the subset of patients with SCN that progresses to AML, acquired mutations in the receptor for granulocyte colony-stimulating factor (G-CSF) have been detected that result in the expression of truncated forms of the G-CSF receptor (G-CSFR) protein. G-CSFR truncation mutants from these patients trans-duce hyperproliferative growth responses. In this paper, we show that the most frequently isolated mutant G-CSFR form from patients with SCN/AML (▵716) confers resistance to apoptosis and prolongs cell survival through a mechanism involving Akt, a downstream target of PI3-kinase. G-CSF stimulation of cells expressing the G-CSFR truncation mutant induces sustained activation of Akt and prolonged phosphorylation of the pro-apoptotic protein Bad, resulting in enhanced cell survival. Extension of cell survival allowing for sufficient time for the acquisition of additional oncogenic events may represent an important mechanism by which G-CSFR mutations contribute to leukemogenesis. These data provide further insight into the pathophysiologic contribution of G-CSFR mutations to AML.
Inv(2)(p23q35) in anaplastic large-cell lymphoma induces constitutive anaplastic lymphoma kinase (ALK) tyrosine kinase activation by fusion to ATIC, an enzyme involved in purine nucleotide biosynthesis
Authors: Ma Zhigui , Cools Jan , Marynen Peter , Cui Xiaoli , Siebert Reiner , Gesk Stefan , Schlegelberger Brigitte , Peeters Benjamin , De Wolf-Peeters Christiane , Wlodarska Iwona , Morris Stephan W.
Published: October 2019

The non-Hodgkin lymphoma (NHL) subtype anaplastic large-cell lymphoma (ALCL) is frequently associated with a t(2;5)(p23;q35) that results in the fusion of the ubiquitously expressed nucleophosmin (NPM) gene at 5q35 to the anaplastic lymphoma kinase (ALK) gene at 2p23, which is not normally expressed in hematopoietic tissues. Approximately 20% of ALCLs that expressALK do not contain the t(2;5), suggesting that other genetic abnormalities can result in aberrant ALK expression. Here we report the molecular characterization of an alternative genetic means of ALK activation, the inv(2)(p23q35). This recurrent abnormality produces a fusion of the amino-terminus of 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC), a bifunctional homodimeric enzyme that catalyzes the penultimate and final steps of de novo purine nucleotide biosynthesis, with the intracellular portion of the ALK receptor tyrosine kinase. RT-PCR analysis of 5 ALCL tumors that contained the inv(2) revealed identical ATIC-ALK fusion cDNA junctions in all of the cases. Transient expression studies show that theATIC-ALK fusion transcript directs the synthesis of an approximately 87-kd chimeric protein that is localized to the cytoplasm, in contrast to NPM-ALK, which typically exhibits a cytoplasmic and nuclear subcellular distribution. ATIC-ALK was constitutively tyrosine phosphorylated and could convert the IL-3–dependent murine hematopoietic cell line BaF3 to cytokine-independent growth. Our studies demonstrate an alternative mechanism for ALK involvement in the genesis of NHL and suggest that ATIC-ALK activation results from ATIC-mediated homodimerization. In addition, expected decreases in ATIC enzymatic function in ATIC-ALK–containing lymphomas may render these tumors more sensitive to antifolate drugs such as methotrexate.
Improved disease-free–survival after transplantation of peripheral blood stem cells as compared with bone marrow from HLA-identical unrelated donors in patients with first chronic phase chronic myeloid leukemia
Authors: Elmaagacli Ahmet H. , Basoglu Semiha , Peceny Rudolf , Trenschel Rudolf , Ottinger Hellmut , Lollert Andre , Runde Volker , Grosse-Wilde Hans , Beelen Dietrich W. , Schaefer Ulrich W.
Published: October 2019

Outcomes after peripheral blood stem cell transplantation (PBSCT) for chronic phase chronic myeloid leukemia (n = 37) were compared with outcomes after bone marrow transplantation (BMT) (n = 54) in the HLA-compatible unrelated donor setting. Median follow-up was 17 months after PBSCT and 29 months after BMT. Both neutrophil and platelet recovery were faster after PBSCT (P < .05). PBSCT was associated with improved immune reconstitution, with higher peripheral blood naive (CD4+CD45RA+) and memory (CD4+ CD45RO+) helper T cells at 3 months and 12 months after transplantation (P < .03). The cumulative incidence of acute (grades II-IV) and chronic graft-versus-host disease (GVHD) were similar, but BMT was associated with a higher cumulative incidence of severe, acute (grade III-IV) GVHD at 24% as compared with 8% with PBSCT (P < .05). Molecular relapse, defined by 2 consecutive positive polymerase chain reaction assays for bcr-abl within a 4-week interval, occurred in 12 of 45 evaluable patients (27%) after BMT and in 4 of 37 (11%) after PBSCT (not significant). Cytogenetic relapse occurred in 5 of 54 patients after BMT (9%) and in 1 of the 37 (3%) patients after PBSCT (not significant). Seventeen of the 54 patients died after BMT (31%), as compared with 2 of 37 patients after PBSCT (5%). Deaths in the BMT group were associated mainly with infections and severe, acute GVHD. The estimated probability of transplant-related mortality (TRM) and disease-free survival at 1000 days after receiving the transplant were 30% and 64% in the BMT group and 5% and 91% in the PBSCT group (P < .03). Overall survival 1000 days after receiving the transplant was 66% after BMT and 94% after PBSCT (P < .02). In the multivariate analysis, only acute GVHD significantly influenced TRM (P < .01).
Sensitization of B-cell chronic lymphocytic leukemia cells to recombinant immunotoxin by immunostimulatory phosphorothioate oligodeoxynucleotides
Authors: Decker Thomas , Hipp Susanne , Kreitman Robert J. , Pastan Ira , Peschel Christian , Licht Thomas
Published: October 2019

A recombinant anti-CD25 immunotoxin, LMB-2, has shown clinical efficacy in hairy cell leukemia and T-cell neoplasms. Its activity in B-cell chronic lymphocytic leukemia (B-CLL) is inferior but might be improved if B-CLL cells expressed higher numbers of CD25 binding sites. It was recently reported that DSP30, a phosphorothioate CpG-oligodeoxynucleotide (CpG-ODN) induces immunogenicity of B-CLL cells by up-regulation of CD25 and other antigens. The present study investigated the antitumor activity of LMB-2 in the presence of DSP30. To this end, B-CLL cells from peripheral blood of patients were isolated immunomagnetically to more than 98% purity. Incubation with DSP30 for 48 hours augmented CD25 expression in 14 of 15 B-CLL samples, as assessed by flow cytometry. DSP30 increased LMB-2 cytotoxicity dose dependently whereas a control ODN with no CpG motif did not. LMB-2 displayed no antitumor cell activity in the absence of CpG-ODN as determined colorimetrically with an (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. In contrast, B-CLL growth was inhibited in 12 of 13 samples with 50% inhibition concentrations (IC50) in the range of LMB-2 plasma levels achieved in clinical studies. Two samples were not evaluable because of spontaneous B-CLL cell death in the presence of DSP30. Control experiments with an immunotoxin that does not recognize hematopoietic cells, and an anti-CD22 immunotoxin, confirmed that sensitization to LMB-2 was specifically due to up-regulation of CD25. LMB-2 was much less toxic to normal B and T lymphocytes compared with B-CLL cells. In summary, immunostimulatory CpG-ODNs efficiently sensitize B-CLL cells to a recombinant immunotoxin by modulation of its target. This new treatment strategy deserves further attention.
Genomic characterization of Haemophilus influenzae: a focus on the capsule locus
Authors: Potts Caelin C. , Topaz Nadav , Rodriguez-Rivera Lorraine D. , Hu Fang , Chang How-Yi , Whaley Melissa J. , Schmink Susanna , Retchless Adam C. , Chen Alexander , Ramos Edward , Doho Gregory H. , Wang Xin
Published: October 2019

Background

Haemophilus influenzae (Hi) can cause invasive diseases such as meningitis, pneumonia, or sepsis. Typeable Hi includes six serotypes (a through f), each expressing a unique capsular polysaccharide. The capsule, encoded by the genes within the capsule locus, is a major virulence factor of typeable Hi. Non-typeable (NTHi) does not express capsule and is associated with invasive and non-invasive diseases.

Methods

A total of 395 typeable and 293 NTHi isolates were characterized by whole genome sequencing (WGS). Phylogenetic analysis and multilocus sequence typing were used to characterize the overall genetic diversity. Pair-wise comparisons were used to evaluate the capsule loci. A WGS serotyping method was developed to predict the Hi serotype. WGS serotyping results were compared to slide agglutination (SAST) or real-time PCR (rt-PCR) serotyping.

Results

Isolates of each Hi serotype clustered into one or two subclades, with each subclade being associated with a distinct sequence type (ST). NTHi isolates were genetically diverse, with seven subclades and 125 STs being detected. Regions I and III of the capsule locus were conserved among the six serotypes (≥82% nucleotide identity). In contrast, genes in Region II were less conserved, with only six gene pairs from all serotypes showing ≥56% nucleotide identity. The WGS serotyping method was 99.9% concordant with SAST and 100% concordant with rt-PCR in determining the Hi serotype.

Conclusions

Genomic analysis revealed a higher degree of genetic diversity among NTHi compared to typeable Hi. The WGS serotyping method accurately predicted the Hi capsule type and can serve as an alternative method for Hi serotyping.
Icotinib-resistant HCC827 cells produce exosomes with mRNA MET oncogenes and mediate the migration and invasion of NSCLC
Authors: Yu Yiming , Abudula Maidinaimu , Li Chaofen , Chen Zhongbo , Zhang Yun , Chen Yichen
Published: October 2019

Background

Icotinib has been widely used in patients with non-small cell lung cancer (NSCLC), and have significantly enhanced the overall survival rate of NSCLC patients. However, acquired drug resistance limits its clinical efficacy. Tumor cell-derived exosomes have been reported to participate in various biological processes, including tumor invasion, metastasis and drug resistance.

Materials and methods

In the present study, drug resistance was measured by MTT assay. Exosomes were extracted from the cell supernatant using ultracentrifugation and identified by exosomal marker. HCC827 cells were treated with exosomes derived from icotinib-resistant (IR) HCC827 to observe the invasion and migration of parent cells. The expression of exo-mRNA was analyzed by reverse transcription-quantitative polymerase chain reaction (RT-PCR). In addition, 10 exo-mRNAs detecting from the plasma and bronchoalveolar lavage fluid (BALF) of NSCLC patients with icotinib treatment were used to establish a new drug resistant-warning formula.

Results

The oncogene MET into exosomes was identified from icotinib-resistant lung cancer cells, and this was also presented in exosomes in NSCLC patients diagnosed with cancer metastasis after icotinib treatment. The knockdown of MET in exosomes significantly decreased the ability of invasion and migration in HCC827 cells.

Conclusion

It was suggested that MET might be specifically package and transferred by exosomes to modify the invasion and migration ability of the surrounding icotinib-sensitive cells.
A hierarchical (multicomponent) model of in-group identification: adaptation of a measure to the Brazilian context
Authors: de Souza Luana Elayne Cunha , de Lima Tiago Jessé Souza , Maia Luciana Maria , Fontenele Ana Beatriz Gomes , Lins Samuel Lincoln Bezerra
Published: October 2019

The aim of this study is to adapt the multidimensional in-group identification scale (MGIS) to the Brazilian context by gathering evidence of its psychometric properties. A total of 663 people from two samples participated in the study. In sample 1, we measured the identification of Brazilians with the region of the country where they live. In sample 2, we measured the identification of students with the university which they attend. Confirmatory factor analyses were performed on both samples to compare the models previously proposed by the original authors of the measure. The obtained results confirmed the validity of the hierarchical and multidimensional factor structure proposed by the original authors. The scale proposed here can be used to measure multiple dimensions of in-group identification in Brazil.
Spermatogenesis disorders in transsexuals
Authors: Kozlov G. I. , Kalinchenko S. Yu. , Slonimsky B. Yu.
Published: October 2019

Examinations of the semen in 8 patients with male transsexualism revealed stereotypical changes in spermograms ranging from oligo- to azoospermia. Possible mechanisms responsible for this abnormality are discussed.
Goiter relapses
Authors: Goch Ye. M. , Kudryashov V. K. , Belyayev P. A.
Published: October 2019

The problem of relapse of goiter is still considered insoluble. Postoperative relapses of goiter are observed quite often - from 1.8 to 39%.

The severity of the problem of recurrent goiter is that at present, patients suffering from this disease can be attributed to the group of "increased oncological risk" in relation to thyroid cancer. Malignancy of recurrent goiter, according to various authors, is observed with a frequency of 8.1 to 20%. Malignancy occurs hidden without clear clinical symptoms of thyroid cancer, and therefore surgical intervention is often performed at a stage that does not give optimal treatment results.

The second side of the problem is the technical features of the surgical treatment of recurrent goiter. The latter are caused by violations of the anatomical and topographic relationships of tissues, extensive scars and atypical location of the node. As a result, complications and mortality after repeated operations with goiter significantly exceed those after primary interventions. Therefore, one of the tasks of thyroid surgery is to find out the causes of relapse of goiter and reduce its frequency.

For this purpose, various methods of primary surgery, the intake of thyroid hormones in the postoperative period, etc. are proposed.

The reasons that lead to relapse of goiter are varied and depend primarily on the disease for which the patient was operated on primarily (diffuse toxic goiter, nodular goiter, multinodal, chronic autoimmune thyroiditis, etc.). To avoid relapse of goiter, a number of surgeons offer more advanced operations even with nodular goiter. However, the expansion of the volume of operation with nodular forms of goiter often leads to the development of persistent hypothyroidism.
Effect of female sex steroids on splenocyte ability to produce an adoptive immune response. Role of prostaglandin F2a in hormonal immunoregulation mechanisms
Authors: Shirshev S. V. , Shilov Yu. I. , Kevorkov N. N.
Published: October 2019

Male CBA mice were used in experiments. Splenocytes were incubated for an hour in macrocultures with estradiol (E2) or progesterone (P), then the cells were transferred (together with the antigen) to lethally irradiated syngeneic recipients, and on day 4 the count of antibody-producing cells (APC) was estimated. Prostaglandin F2a (PGF2a) concentrations were radioimmunoassayed in cell culture supernatants. E2 and P concentrations corresponded to these hormones levels in blood sera during pregnancy. E2 and P in the tested concentrations were found to reliably stimulate the processes of APC formation, their effects being dose independent. Both E2 and P statistically reliably increased PGF2a level in splenocyte culture. Fractionation of splenocytes helped reveal the highest PGF2a level in the cultures devoid of adhesive cells and rich for T lymphocytes. Hence, E2 and P stimulated the processes of antigen-independent differentiation of splenocytes producing APC either by directly stimulating T lymphocytes or via macrophages by blocking their negative effects. Stimulation of adoptive immune response by sex steroids is closely connected with PGF2a production by immunocompetent cells under the effects of these hormones.
The effect of new long-acting testosterone bucyclate ester on the spermatogenesis and endocrine function of steroid-secreting glands in male Papio hamadryas
Authors: Goncharov N. P. , Katsiya G. V. , Butnev V. Yu. , Gorlushkin V. M.
Published: October 2019

Effects of two (at 3 month interval) intramuscular injections of testosterone bucyclate (ТВ) in doses of 4 and 8 mg/kg b. w. were studied in adult male Papio hamadryas. Both ТВ doses suppressed spermatogenesis, but this effect abated with dose increase. Azoospermia was observed in 33% ejaculate samples after ТВ in a dose of 4 mg/kg, whereas after injection of 8 mg/kg it was found in only 6% samples. Moreover, the level of peripheral testosterone depends on ТВ dose as well. It reduces by half after injection of a lower dose and surpasses the control values by 20 to 120% after injection of 8 mg/kg. Time course of LH after injection of the androgen in both doses was characterized by a phase pattern: during the first phase (0 to 16 weeks) the levels of hormone varied, and a trend to reduction could be traced; during the second phase (weeks 16-24) the concentration of the hormone sharply increased and thus remained till the end of experiment. Prolonged exposure to long-acting ТВ may inhibit the secretion of adrenal androgen dehydroepiandrosterone and precursor of the ∆5-pathway of steroid synthesis 17-Oxypregnelone.
The case of histiocytosis X (Hand - Schüller – Christian disease) with thyroid gland lesion
Authors: Bronstein M. E. , Melnichenko G. A. , Bukhman A. I. , Starostina T. A. , Ivanov O. L. , Pronin V. S. , Pavlova M. G. , Oranskaya A. N.
Published: October 2019

Histiocytosis X, which is a rare disease, combines 3 related but different in clinical manifestation, course and prognosis of the syndrome: eosinophilic granuloma (Taratynovs disease), Hand Schller Christian disease (syndrome) and Letterer Siw disease (syndrome). The clinical picture of Hend Schller Christian disease was first described by Hand in 1893, then by Schller in 1915 and Christian in 1920. The term histiocytosis X was first proposed in 1953 by Liechtenstein, who combined all 3 of the aforementioned diseases under this name. on the basis of the similarity of the clinical picture, the possibility of a mutual transition from one state to another and typical morphological manifestations consisting in the proliferation of histiocytes and the accumulation of various lipids in their cytoplasm. In 1965, this concept was confirmed in all three diseases, there are no special pentalaminar markers in the cytoplasm of histiocytes that are detected in Langerhans cells (hence the name of these inclusions - "Langerhans cell granules"), the origin and biological significance of which remains unclear. Histiocytes have no signs of atypia, although they infiltrate many organs and tissues retain their normal properties. Histiocytic proliferation can develop in various organs and tissues, causing them to corresponding functional and morphological changes.
Tourist tongues: High-speed rail carries linguistic and cultural urbanisation beyond the city limits in Guangxi, China
Authors: Grey Alexandra
Published: October 2019

This article builds upon research which analyses the reconstruction of cities in China as an integral part of image-making discourses competing to attract mobile capital. It extends that literature beyond urban places to urbanisation processes, examining the material and linguistic features of networks and discourses of new high-speed rail infrastructure Guangxi, a poorer, rural, multilingual and multiethnic region of the People’s Republic of China (China) in which tourism – propelled by high-speed trains – has become a pillar of economic development. It argues that these trains produce symbolically powerful discourses which contribute to cultural urbanisation across Guangxi, emplacing urban norms outside city limits in pursuit of profitable sameness, as tourism does not trade only upon difference. Local multilingualism, specifically, is erased as too different, a barrier to tourists’ (and tourism capital’s) mobility. Amongst other ramifications, this reproduces social distance and ideologically displaces local languages.
The Impact of Hydrometeorological Events on the State of the Urban River Quality
Authors: Pietruszyński Łukasz , Cieśliński Roman
Published: October 2019

One of the aims of this paper was to study catchment and watercourse response to extreme events. The study area consisted of Orunski Stream in the northern Polish city of Gdansk. The study period (review of cartographic materials) covered the years 1908 to 2000, while field research covered the period from May 2009 to April 2010. In addition, measurements were performed to study the dynamics of changes occurring in the creek during rainfall events. Examination of the water chemistry changes made two days before heavy rainfall showed a significant increase in sulphates and nitrogen compounds. Additionally, pollutants washed from the basin contributed to a significant deterioration in the water colour. A rainfall of 1.3 mm caused an eleven-fold increase in stream flow, a twofold increase in the water level, and a fourfold increase in the velocity of water molecules. It might appear seem that such a small amount of rainfall would have no major effect on the hydrologic and hydrochemical parameters of a small creek. However, it is the maximum flow rate that is an important variable when assessing potential flood risk levels.
The Influence of the Filtration Bed type in the Pool Water Treatment System on Washings Quality
Authors: Wyczarska-Kokot Joanna , Lempart Anna
Published: October 2019

This paper presents the influence of the type of filtration beds, used in swimming pool water treatment systems, on the quality and the possibility of reuse of washings. The research covered 4 pool cycles with sand, sand and anthracite, glass and diatomaceous beds. The degree of contamination of washings was assessed on the basis of physical, chemical and bacteriological tests. The possibility of washings drainage into the natural environment was considered, and the results of the research were compared with the permissible values of pollution indicators for wastewater discharged to water or ground. A direct management of washings from the analysed filters proved impossible mainly due to the high content of TSS (total suspended solids) and free chlorine. Washings were subjected to sedimentation and then the supernatant was stirred intensively. As a result of these processes, the quality of washings was significantly improved. This allowed planning to supplement the pool water installations with systems for washings management.
Effect of Municipal Sewage Sludge on Soil Chemical Properties and Chemical Composition of Spring Wheat
Authors: Antonkiewicz Jacek , Kuc Andrzej , Witkowicz Robert , Tabak Monika
Published: October 2019

Municipal sewage sludge from rural sewage treatment plants is characterized by a substantial content of organic matter and macronutrients, which can be used in cultivation of cereals. In a farm located in the commune of Iwanowice in the south of Poland (Malopolska province), municipal sewage sludge was applied under spring wheat cultivation. The experiment was set up on heavy soil with slightly acid reaction and medium content of available forms of P, K, Mg. Application of sewage sludge in a dose of 23 Mg fresh matter per hectare (4.21 Mg d.m.) led to no significant changes in chemical properties of the soil. Application of sewage sludge significantly increased yield of spring wheat. That increase led to a significant decrease in the content of N, P, K, Na, Mg and Ca in spring wheat. Utilization of N, Mg, K, P and Ca from sewage sludge by spring wheat was at a level of 82, 63, 44, 36, 9 %, respectively, of the amount introduced with the waste. Application of municipal sewage sludge significantly decreased the value of Ca : P ratio in spring wheat grain and straw. The sewage sludge did not cause a significant change in the values of Ca : Mg, K : Na, K : (Ca+Mg), K : Mg and K : Ca ratios in spring wheat grain and straw. Municipal sewage sludge can be used environmentally, including for fertilization of cereals, provided that environmental standards are kept.
Agronomic performances of aromatic and non-aromatic M1 rice under drought stress
Authors: Herwibawa Bagus , Sakhidin  , Haryanto Totok Agung Dwi
Published: October 2019

Flooded rice paddy fields act as a source of greenhouse gas emissions. So, the efforts to increase the drought tolerance of rice represent a much more environmentally friendly solution, and may bring a significant contribution to prevent global warming. The experiment was laid out in a split-plot design with ten replicates. Rice cultivars treated with different levels of γ irradiation and sodium azide (SA) soaking time were allocated in the main plots, and mutagen groups in sub-plots. We use the generalized linear models, as implemented in the GLIMMIX procedure of SAS University Edition, to analyze agronomic performances. These results showed that the genotypes in M1 generation present diversity under the drought stress level of -0.03 MPa, and the best combination between cultivar and mutagen is Inpago Unsoed 1 that was irradiated with γ 100 Gy and then soaked in SA for 2h. These genotypes can be used as a sources of drought tolerance in future rice breeding programmes.
Mesenchymal stem cells as a treatment for multiple sclerosis: a focus on experimental animal studies
Authors: Lotfy Ahmed , Ali Nourhan S. , Abdelgawad Mai , Salama Mohamed
Published: October 2019

Multiple sclerosis (MS) is a progressive and debilitating neurological condition in which the immune system abnormally attacks the myelin sheath insulating the nerves. Mesenchymal stem cells (MSCs) are found in most adult tissues and play a significant systemic role in self-repair. MSCs have promising therapeutic effects in many diseases, such as autoimmune diseases, including MS. MSCs have been tested in MS animal models, such as experimental autoimmune encephalomyelitis. Other studies have combined other agents with MSCs, genetically modified MSCs, or used culture medium from MSCs. In this review, we will summarize these studies and compare the main factors in each study, such as the source of MSCs, the type of animal model, the route of injection, the number of injected cells, and the mechanism of action.