bluuebunny/crossref_metadata_2025 huggingface dataset:
The incidence of clonal T-cell receptor rearrangements in B-cell precursor acute lymphoblastic leukemia varies with age and genotype
Authors: Brumpt Caren , Delabesse Eric , Beldjord Kheira , Davi Frederic , Cayuela Jean-Michel , Millien Corinne , Villarese Patrick , Quartier Pierre , Buzyn Agnes , Valensi Françoise , Macintyre Elizabeth
Published: October 2019

B-cell precursor acute lymphoblastic leukemias (BCP-ALLs) are increasingly treated on risk-adapted protocols based on presenting clinical and biological features. Residual molecular positivity of clonal immunoglobulin (IG) and T-cell receptor (TCR) rearrangements allows detection of patients at an increased risk of relapse. If these rearrangements are to be used for universal follow-up, it is important to determine the extent to which they are informative in different BCP-ALL subsets. We show thatIGH V-D-J rearrangements occur in 89% of 163 BCP-ALL, with no significant variation according to age or genotype (BCR-ABL, TEL-AML1, MLL-AF4, and E2A-PBX1). In contrast,TCRG rearrangements, which occur in 60% of patients overall, are frequent in BCR-ABL and TEL-AML1, are less so in MLL-AF4, and are virtually absent in infants aged predominantly from 1 to 2 years and in E2A-PBX1 ALLs. Incidence of the predominant TCRD Vδ2-Dδ3 rearrangement decreases with age but is independent of genotype. These differences are not due to differential recombination activating gene activity, nor can they be explained adequately by stage of maturation arrest. Analysis of MLL-AF4 BCP-ALL is in keeping with transformation of a precursor at an early stage of ontogenic development, despite the adult onset of the cases analyzed. We postulate that the complete absence of TCRG rearrangement in E2A-PBX1 cases may result from deregulated E2A function. These data also have practical consequences for the use ofTCR clonality for the molecular follow-up of BCP-ALL.
Synergism of multiple adhesion molecules in mediating cytoadherence of Plasmodium falciparum–infected erythrocytes to microvascular endothelial cells under flow
Authors: Yipp Bryan G. , Anand Samantha , Schollaardt Tineke , Patel Kamala D. , Looareesuwan Sornchai , Ho May
Published: October 2019

Plasmodium falciparum–infected erythrocytes (IRBCs) have been shown to interact with a number of endothelial adhesion molecules expressed on transfectants, on cell lines, and as immobilized purified receptor proteins under flow conditions. However, the experiments were designed in such a way that maximal numbers of adhesion molecules were provided as substratum. Whether the interactive events actually occur on microvascular endothelium, where the distribution and expression of adhesion molecules may be less, remains undetermined. In this study, the cytoadherance of IRBCs on human dermal microvascular endothelial cells (HDMECs) as a model of human microvasculature was examined. IRBCs were observed to tether, roll, and adhere on resting HDMECs, which constitutively expressed CD36 and intercellular adhesion molecule-1 (ICAM-1) at an optimal shear stress of 1 dyne/cm2. Stimulation of HDMECs with tumor necrosis factor–α for 5 and 24 hours, which resulted in up-regulation of ICAM-1 and induction of vascular cell adhesion molecule-1 expression, significantly increased the percentage of rolling cells that adhered without affecting the rolling flux. In contrast, P-selectin expression on HDMECs induced by oncostatin M led to an increase in both rolling flux and adhesion. Inhibition studies with receptor-specific monoclonal antibodies revealed that adhesion of IRBCs on HDMECs was largely CD36 dependent, whereas rolling could be mediated by any of the adhesion molecules studied. Collectively, these findings indicate that IRBCs interact synergistically with multiple adhesion molecules on vascular endothelium. The rolling of IRBCs may be the rate-limiting step in cytoadherance, since it can be modulated by cytokines to enhance CD36-mediated IRBC adhesion.
Antihirudin antibodies in patients with heparin-induced thrombocytopenia treated with lepirudin: incidence, effects on aPTT, and clinical relevance
Authors: Eichler Petra , Friesen Heinz-Juergen , Lubenow Norbert , Jaeger Bernd , Greinacher Andreas
Published: October 2019

Hirudin, a potent and specific thrombin inhibitor, is a protein of nonhuman origin and therefore potentially immunogenic. The primary objectives of this investigation were to determine the incidence of antihirudin antibodies (ahir-ab) in patients with heparin-induced thrombocytopenia (HIT) who received lepirudin as parenteral anticoagulation and to determine the incidence of death, limb amputation, new thromboembolic complications (TECs), and major hemorrhage in patients who had ahir-ab, compared with patients who were ahir-ab negative. The investigation used data from 2 prospective multicenter studies with the same study protocol, in which HIT patients received 1 of 4 intravenous lepirudin dosage regimens. The treatment duration was 2 to 10 days. Ahir-ab were determined by a newly developed enzyme-linked immunosorbent assay (ELISA). Eighty-seven of 196 evaluable patients (44.4%) had ahir-ab of the IgG class. Development of ahir-ab was dependent on the duration of treatment (ahir-ab–positive patients 18.6 days vs ahir-ab–negative patients 11.8 days; P = .0001). Fewer ahir-ab–positive than ahir-ab–negative patients died (P = .001). Ahir-ab did not cause an increase in limb amputation (P = .765), new TECs (P > .99), or major bleedings (P = .549). In 23 of 51 (45.1%) evaluable patients in whom ahir-ab developed during treatment with lepirudin ( = 12% of all lepirudin treated patients), the ahir-ab enhanced the anticoagulatory effect of lepirudin. Ahir-ab are frequent in patients treated with lepirudin for more than 5 days. Ahir-ab are the first example for a drug-induced immune response causing enhanced activity of a drug. Therefore, during prolonged treatment with lepirudin, anticoagulatory activity should be monitored daily to avoid bleeding complications.
Efficient retrovirus-mediated gene transfer to transplantable human bone marrow cells in the absence of fibronectin
Authors: Hennemann Burkhard , Oh Il-Hoan , Chuo Jean Y. , Kalberer Christian P. , Schley Patricia D. , Rose-John Stefan , Humphries R. Keith , Eaves Connie J.
Published: October 2019

The low frequency of transplantable hematopoietic stem cells in adult human bone marrow (BM) and other differences from cord blood stem cells have impeded studies to optimize the retroviral transduction of stem cells from adult sources. To address this problem, first a cytokine combination was defined that would both maximize the kinetics of adult BM CD34+CD38− cell mitogenesis and minimize the period of prestimulation required for the transduction of these cells by a MSCV-GFP/neor virus in tissue culture dishes in the absence of fibronectin. Three days of stimulation with flt3-ligand, Steel factor, interleukin (IL)-3, and hyper-IL-6 proved both necessary and sufficient to obtain 83% ± 2% GFP+ CD34+CD38− cells, 75% ± 10% G418-resistant clonogenic progenitors, and 50% ± 20% transduced long-term culture-initiating cells as recovered 48 hours after a single exposure to virus. Moreover, this was accompanied by a several-fold increase in viral receptor (pit-1) messenger RNA transcripts in the target cells. Using this prestimulation protocol, repeated daily exposure to new virus (3×) did not alter the proportion of transduced cells over that obtained with a single exposure. Adult human BM cells able to engraft immunodeficient (NOD/SCID-β2M−/−) mice were also efficiently transduced (10%-20% GFP+ human lymphoid and myeloid cells present 6-8 weeks after transplant) using a 6-day prestimulation and infection protocol. A clinically useful efficiency of retrovirus-mediated gene transfer to transplantable adult human BM stem cells can thus be obtained with a protocol that allows their semisynchronous activation into cycle and concomitant increased expression of virus receptor transcripts before virus exposure.
Recruitment of the nuclear receptor corepressor N-CoR by the TEL moiety of the childhood leukemia–associated TEL-AML1 oncoprotein
Authors: Guidez Fabien , Petrie Kevin , Ford Anthony M. , Lu Huafeng , Bennett Caroline A. , MacGregor Angus , Hannemann Jürgen , Ito Yoshiaki , Ghysdael Jacques , Greaves Mel , Wiedemann Leanne M. , Zelent Arthur
Published: October 2019

The t(12;21)(p13;q22) chromosomal translocation is the most frequent illegitimate gene recombination in a pediatric cancer and occurs in approximately 25% of common acute lymphoblastic leukemia (cALL) cases. This rearrangement results in the in frame fusion of the 5′-region of the ETS-related gene, TEL(ETV6), to almost the entire acute myeloid leukemia 1 (AML1) (also called CBFA2 orPEBP2AB1) locus and expression of the TEL-AML1 chimeric protein. Although AML1 stimulates transcription, TEL-AML1 functions as a repressor of some AML1 target genes. In contrast to the wild type AML1 protein, both TEL and TEL-AML1 interact with N-CoR, a component of the nuclear receptor corepressor complex with histone deacetylase activity. The interaction between TEL and N-CoR requires the central region of TEL, which is retained in TEL-AML1, and TEL lacking this domain is impaired in transcriptional repression. Taken together, our results suggest that TEL-AML1 may contribute to leukemogenesis by recruiting N-CoR to AML1 target genes and thus imposing an altered pattern of their expression.
Durability of responses following donor lymphocyte infusions for patients who relapse after allogeneic stem cell transplantation for chronic myeloid leukemia
Authors: Dazzi Francesco , Szydlo Richard M. , Cross Nicholas C. P. , Craddock Charles , Kaeda Jaspal , Kanfer Ed , Cwynarski Kate , Olavarria Eduardo , Yong Agnes , Apperley Jane F. , Goldman John M.
Published: October 2019

An analysis was performed of the response to treatment with donor lymphocyte infusions (DLI) and the survival in 66 consecutive patients who relapsed after primary treatment by allogeneic stem cell transplantation for BCR-ABL–positive chronic myeloid leukemia. The transplant donor was an HLA-identical sibling (n = 35) or a “matched” unrelated volunteer (n = 31). Fifty-seven patients were transplanted in chronic phase, eight in accelerated phase, and one in second chronic phase. The recognition of relapse was based on precise molecular, cytogenetic, or hematologic criteria. The median interval from transplant to relapse was 12 months (range 3-85). The median interval from relapse to initiation of DLI was 9.4 months (range 1-70). Patients received DLI from their original transplant donors on a bulk-dose (n = 34) or on an escalating-dose (n = 32) regimen. Patients were monitored serially by hematologic, cytogenetic, and molecular criteria. Molecular remission was defined by the finding of negative results by nested primer reverse transcriptase polymerase chain reaction (RT-PCR) for BCR-ABL transcripts on two consecutive occasions, subject to satisfactory controls. Forty-four patients (67%) achieved molecular remission. Patients who had relapsed to advanced phase disease and patients with short intervals between transplant and relapse had significantly lower probabilities of achieving molecular remission. Of the 44 patients who achieved molecular remission, 4 reverted to a PCR-positive status at 15, 18, 37, and 87 weeks after remission. The probability of survival for patients who achieved molecular remission was significantly better than for those who failed to do so (95% versus 53% at 3 years post-DLI,P = .0001). We conclude that the majority of molecular remissions after DLI are durable, and thus the majority of responding patients may prove to have been cured.
Rapid reconstitution of Epstein-Barr virus–specific T lymphocytes following allogeneic stem cell transplantation
Authors: Marshall Natalie A. , Howe John Greg , Formica Richard , Krause Diane , Wagner John E. , Berliner Nancy , Crouch Jill , Pilip Ingrid , Cooper Dennis , Blazar Bruce R. , Seropian Stuart , Pamer Eric G.
Published: October 2019

Epstein-Barr virus (EBV)–specific CD8 T lymphocytes are present at remarkably high frequencies in healthy EBV+individuals and provide protection from EBV-associated lymphoproliferative diseases. Allogeneic peripheral blood stem cell transplantation (allo-PBSCT) is a commonly used therapy in which T-cell surveillance for EBV is temporarily disrupted. Herein, human leukocyte antigen (HLA) class I tetramers were used to investigate the reestablishment of the EBV-specific CD8 T-cell repertoire in patients following allo-PBSCT. CD8+ T cells specific for lytic and latent cycle–derived EBV peptides rapidly repopulate the periphery of matched sibling allo-PBSCT patients. The relative frequencies of T cells specific for different EBV peptides in transplantation recipients closely reflect those of their respective donors. Investigation of patients at monthly intervals following unmanipulated allo-PBSCT demonstrated that the frequency of EBV-specific T cells correlates with the number of EBV genome copies in the peripheral blood and that expansion of EBV-specific T-cell populations occurs even in the setting of immunosuppressive therapy. In contrast, patients undergoing T-cell–depleted or unrelated cord blood transplantation have undetectable EBV-specific T cells, even in the presence of Epstein-Barr viremia. The protective shield provided by EBV-specific CD8 T cells is rapidly established following unmanipulated matched sibling allo-PBSCT and demonstrates that HLA class I tetramers complexed with viral peptides can provide direct and rapid assessment of pathogen-specific immunity in this and other vulnerable patient populations.
CD40-ligand (CD154) gene therapy for chronic lymphocytic leukemia
Authors: Wierda William G. , Cantwell Mark J. , Woods Sandra J. , Rassenti Laura Z. , Prussak Charles E. , Kipps Thomas J.
Published: October 2019

Chronic lymphocytic leukemia (CLL) cells can be made to express recombinant CD40-ligand (CD154) by transduction with a replication-defective adenovirus vector (Ad-CD154). Ad-CD154–transduced and bystander leukemia cells become highly effective antigen-presenting cells that can induce CLL-specific autologous cytotoxic T lymphocytes in vitro. This study investigated the immunologic and clinical responses to infusion of autologous Ad-CD154-CLL cells in patients with CLL. After a one-time bolus infusion of autologous Ad-CD154–transduced leukemia cells, there was increased or de novo expression of immune accessory molecules on bystander, noninfected CLL cells in vivo. Treated patients also developed high plasma levels of interleukin-12 and interferon-γ, the magnitudes of which corresponded to absolute blood CD4+T-cell counts before therapy. On average, patients experienced a greater than 240% increase in absolute blood T-cell counts within 1 to 4 weeks of treatment. Moreover, treatment increased the numbers of leukemia-specific T cells, demonstrated by autologous ELISPOT assay and mixed lymphocyte reactions. These biologic effects were associated with reductions in leukemia cell counts and lymph node size. Treatment did not induce autoimmune thrombocytopenia or hemolytic anemia and no dose-limiting toxicity was observed. This approach may provide a novel and effective form of gene therapy for patients with this disease.
A minimal c-fes cassette directs myeloid-specific expression in transgenic mice
Authors: Heydemann Ahlke , Warming Soren , Clendenin Cynthia , Sigrist Kirsten , Hjorth J. Peter , Simon M. Celeste
Published: October 2019

The c-fes proto-oncogene encodes a 92-kd protein tyrosine kinase whose expression is restricted largely to myeloid and endothelial cells in adult mammals. A 13.2-kilobase (kb) humanc-fes genomic fragment was previously shown to containcis-acting element(s) sufficient for a locus control function in bone marrow macrophages. Locus control regions (LCRs) confer transgene expression in mice that is integration site independent, copy number dependent, and similar to endogenous murine messenger RNA levels. To identify sequences required for this LCR,c-fes transgenes were analyzed in mice. Myeloid-cell–specific, deoxyribonuclease-I–hypersensitive sites localized to the 3′ boundary of exon 1 and intron 3 are required to confer high-level transgene expression comparable to endogenous c-fes, independent of integration site. We define a minimal LCR element as DNA sequences (nucleotides +28 to +2523 relative to the transcription start site) located within intron 1 to intron 3 of the human locus. When this 2.5-kb DNA fragment was linked to a c-fes complementary DNA regulated by its own 446–base-pair promoter, integration-site–independent, copy-number–dependent transcription was observed in myeloid cells in transgenic mice. Furthermore, this 2.5-kb cassette directed expression of a heterologous gene (enhanced green fluorescent protein) exclusively in myeloid cells. The c-fes regulatory unit represents a novel reagent for targeting gene expression to macrophages and neutrophils in transgenic mice.
Smooth muscle cell surface tissue factor pathway activation by oxidized low-density lipoprotein requires cellular lipid peroxidation
Authors: Penn Marc S. , Cui Mei-Zhen , Winokur Allison L. , Bethea John , Hamilton Thomas A. , DiCorleto Paul E. , Chisolm Guy M.
Published: October 2019

Tissue factor, which is expressed in vascular lesions, increases thrombin production, blood coagulation, and smooth muscle cell proliferation. We demonstrate that oxidized low-density lipoprotein (LDL) induces surface tissue factor pathway activity (ie, activity of the tissue factor:factor VIIa complex) on human and rat smooth muscle cells. Tissue factor messenger RNA (mRNA) was induced by oxidized LDL or native LDL; however, native LDL did not markedly increase tissue factor activity. We hypothesized that oxidized LDL mediated the activation of the tissue factor pathway via an oxidant-dependent mechanism, because antioxidants blocked the enhanced tissue factor pathway activity by oxidized LDL, but not the increased mRNA or protein induction. We separated total lipid extracts of oxidized LDL using high-performance liquid chromatography (HPLC). This yielded 2 major peaks that induced tissue factor activity. Of the known oxysterols contained in the first peak, 7α- or 7β-hydroxy or 7-ketocholesterol had no effect on tissue factor pathway activity; however, 7β-hydroperoxycholesterol increased tissue factor pathway activity without induction of tissue factor mRNA. Tertiary butyl hydroperoxide also increased tissue factor pathway activity, suggesting that lipid hydroperoxides, some of which exist in atherosclerotic lesions, activate the tissue factor pathway. We speculate that thrombin production could be elevated via a mechanism involving peroxidation of cellular lipids, contributing to arterial thrombosis after plaque rupture. Our data suggest a mechanism by which antioxidants may offer a clinical benefit in acute coronary syndrome and restenosis.
In vitro and in vivo production of vascular endothelial growth factor by chronic lymphocytic leukemia cells
Authors: Chen Haijuan , Treweeke Andy T. , West Dave C. , Till Kathleen J. , Cawley John C. , Zuzel Mirko , Toh Cheng H.
Published: October 2019

Expansion of primary solid tumors and their malignant dissemination are angiogenesis-dependent. Vascular endothelial growth factor (VEGF) is the key factor playing a pivotal role in solid tumor-induced angiogenesis. Recent studies indicate that angiogenesis may also be involved in the pathogenesis of certain hemic malignancies, including B-cell chronic lymphocytic leukemia (B-CLL). Mechanisms underlying angiogenesis in B-CLL and the role of VEGF in this process are incompletely understood. In this study, it was examined whether angiogenically functional VEGF is produced by B-CLL cells. Immunohistochemical staining with antibodies against VEGF and CD34, an endothelial cell marker, demonstrated the presence of VEGF protein and abundant blood vessels in infiltrated lymphoreticular tissues. Low levels of VEGF were detected by ELISA in the culture media of unstimulated cells; this was enhanced up to 7-fold by hypoxic stimulation. SDS-PAGE and Western blot analysis of the concentrated culture media showed 2 isoforms of VEGF protein with molecular weights of 28 and 42 kd, respectively. RNA hybridization showed that these cells expressed VEGF mRNA. Reverse transcription–polymerase chain reaction, combined with nucleotide sequence analysis, revealed that the predominantly expressed isoforms were VEGF121 and VEGF165. Moreover, 3H-thymidine incorporation and an in vivo angiogenic assay demonstrated that the VEGF produced by CLL cells can induce angiogenesis by stimulating endothelial cell proliferation. In conclusion, this study shows that B-CLL cells produce VEGF and demonstrates the angiogenic effects of this growth factor, which may be relevant for the tissue phase of the disease.
Kaposi sarcoma is a therapeutic target for vitamin D3receptor agonist
Authors: Masood Rizwan , Nagpal Sunil , Zheng Tong , Cai Jie , Tulpule Anil , Smith D. Lynne , Gill Parkash S.
Published: October 2019

Kaposi sarcoma (KS) is responsive to a number of different steroid hormones, such as glucocorticoids and retinoids. An active metabolite of vitamin D, 1α,25 dihydroxyvitamin D3, was used to study the effect of this steroid hormone in KS. Steroid hormones exert their effect through their cognate nuclear receptors, which for vitamin D metabolites is the vitamin D receptor (VDR). It was first shown that KS cell lines and primary tumor tissue express high levels of VDR, whereas endothelial cells had minimal expression and fibroblasts had no expression. Second, KS cell growth was inhibited by VDR agonist 1α,25 dihydroxyvitamin D3 with a 50% inhibitory concentration of 5 × 10 −8 mol/L, whereas endothelial cells and fibroblast cells showed no response. Studies on the mechanism of KS tumor growth inhibition by 1α,25 dihydroxyvitamin D3 showed that production of autocrine growth factors interleukin (IL)-6 and IL-8 was reduced in a dose-dependent manner, whereas no effect was observed on vascular endothelial growth factor and basic fibroblast growth factor. Transcription initiated at the IL-6 promoter was repressed by VDR agonist. The DNA sequences required to mediate this repression were localized to nucleotides −225/−110 in the 5′-flanking region. The antitumor activity of VDR agonists was also confirmed in KS tumor xenograft and after topical application in patients with KS. 1α,25 Dihydroxyvitamin D3 and its analogs may thus be candidates for clinical development in KS.
Molecular evidence of organ-related transmission of Kaposi sarcoma–associated herpesvirus or human herpesvirus-8 in transplant patients
Authors: Luppi Mario , Barozzi Patrizia , Santagostino Gaia , Trovato Raffaella , Schulz Thomas F. , Marasca Roberto , Bottalico Davide , Bignardi Lucia , Torelli Giuseppe
Published: October 2019

In transplant patients, Kaposi sarcoma (KS)-associated herpesvirus or human herpesvirus-8 (HHV-8) infection is associated with the development of KS, primary effusion lymphoma and Castleman disease. Whether HHV-8 is either reactivated in the recipient or transmitted by the donor has been investigated so far only by serologic studies. Thus, we addressed the issue of HHV-8 transmission in the transplantation setting by molecular methods. We exploited the high level variability of the orf-K1 gene and the polymorphism of theorf-73 gene of the HHV-8 genome to assess the genetic relatedness of the HHV-8 strains identified in the posttransplant KS lesions that developed, simultaneously, 20 months after transplantation, in 2 recipients of twin kidneys from the same cadaver donor. The 100% identity of nucleotide sequence of the most variable viral region and the presence of the same, singleorf-73 type in both patients provides strong molecular evidence of organ-related transmission of HHV-8 in the setting of transplantation.
Melphalan and purine analog–containing preparative regimens: reduced-intensity conditioning for patients with hematologic malignancies undergoing allogeneic progenitor cell transplantation
Authors: Giralt Sergio , Thall Peter F. , Khouri Issa , Wang Xuemei , Braunschweig Ira , Ippolitti Cindy , Claxton David , Donato Michele , Bruton Jill , Cohen Agueda , Davis Marilyn , Andersson Borje S. , Anderlini Paolo , Gajewski James , Kornblau Steven , Andreeff Michael , Przepiorka Donna , Ueno Naoto T. , Molldrem Jeff , Champlin Richard
Published: October 2019

A reduced-intensity preparative regimen consisting of melphalan and a purine analog was evaluated for allogeneic transplantation in 86 patients who had a variety of hematologic malignancies and were considered poor candidates for conventional myeloablative therapies because of age or comorbidity. Seventy-eight patients received fludarabine 25 mg/m2 daily for 5 days in combination with melphalan 180 mg/m2 (n = 66) or 140 mg/m2 (n = 12). Eight patients received cladribine 12 mg/m2 continuous infusion for 5 days with melphalan 180 mg/m2. The median age was 52 years (range, 22-70 years). Disease status at transplantation was either first remission or first chronic phase in 7 patients, untreated first relapse or subsequent remission in 16 patients, and refractory leukemia or transformed chronic myelogenous leukemia in 63 patients. Nonrelapse mortality rates on day 100 were 37.4% for the fludarabine/melphalan combination and 87.5% for the cladribine/melphalan combination. The median percentage of donor cells at 1 month in 75 patients was 100% (range, 0%-100%). The probability of grade 2-4 and 3-4 acute graft-versus-host disease was 0.49 (95% CI, 0.38-0.60) and 0.29 (95% CI, 0.18-0.41), respectively. Disease-free survival at 1 year was 57% for patients in first remission or chronic phase and 49% for patients with untreated first relapse or in a second or later remission. On multivariate analysis the strongest predictor for disease-free survival was a good or intermediate risk category. In summary, fludarabine/melphalan combinations are feasible in older patients with associated comorbidities, and long-term disease control can be achieved with reduced-intensity conditioning in this population.
Morphologic and functional characterization of granulocytes and macrophages in embryonic and adult zebrafish
Authors: Lieschke Graham J. , Oates Andrew C. , Crowhurst Meredith O. , Ward Alister C. , Layton Judith E.
Published: October 2019

The zebrafish is a useful model organism for developmental and genetic studies. The morphology and function of zebrafish myeloid cells were characterized. Adult zebrafish contain 2 distinct granulocytes, a heterophil and a rarer eosinophil, both of which circulate and are generated in the kidney, the adult hematopoietic organ. Heterophils show strong histochemical myeloperoxidasic activity, although weaker peroxidase activity was observed under some conditions in eosinophils and erythrocytes. Embryonic zebrafish have circulating immature heterophils by 48 hours after fertilization (hpf). A zebrafish myeloperoxidase homologue (myeloid-specificperoxidase; mpx) was isolated. Phylogenetic analysis suggested it represented a gene ancestral to the mammalian myeloperoxidase gene family. It was expressed in adult granulocytes and in embryos from 18 hpf, first diffusely in the axial intermediate cell mass and then discretely in a dispersed cell population. Comparison of hemoglobinized cell distribution,mpx gene expression, and myeloperoxidase histochemistry in wild-type and mutant embryos confirmed that the latter reliably identified a population of myeloid cells. Studies in embryos after tail transection demonstrated that mpx- and peroxidase-expressing cells were mobile and localized to a site of inflammation, indicating functional capability of these embryonic granulocytes. Embryonic macrophages removed carbon particles from the circulation by phagocytosis. Collectively, these observations have demonstrated the early onset of zebrafish granulopoiesis, have proved that granulocytes circulate by 48 hpf, and have demonstrated the functional activity of embryonic granulocytes and macrophages. These observations will facilitate the application of this genetically tractable organism to the study of myelopoiesis.
Optimization of a simple, accurate and low cost method for starch quantification in green microalgae
Authors: Yong Tze Ching , Chiu Chia-Sheng , Chen Ching-Nen Nathan
Published: October 2019

Background

Lipids and starch are important feedstocks for bioenergy production. Genetic studies on the biosyntheses of lipids and starch in green microalgae have drawn significant attention recently. In these studies, quantifications of lipids and starch are required to clarify the causal effects. While lipids are assayed with similar procedures worldwide, starch in green microalgae has been measured using various methods with deficiencies in accuracy or high cost.

Results

A simple, accurate and low cost procedure for routine quantification of starch in green microalgae was developed. This procedure consists of quick-freezing of the cells, solvent extraction of the pigments, 134 °C autoclaving and glucoamylase double digestions of starch, followed by a glucose assay using the dinitrosalicylic acid reagent. This procedure was optimized to quantify starch in small volumes of green microalgal culture. The accuracy of starch quantification using this procedure was 102.3 ± 2.5% (mean ± SD, n = 6), as indicated by using cornstarch as internal controls. The working protocol is available athttp://dx.doi.org/10.17504/protocols.io.2mhgc36.

Conclusions

This quantification approach overcomes the current problems in the starch quantification of green microalgae such as inaccuracy and high cost. This approach would provide an opportunity to compare the effects of genetic, physiological or cultivation manipulations on the productivity of starch in green microalgae elucidated in different labs, which is essential in the enhancement of lipid productivity studies in microalgae.
Choroidal granuloma caused by Mycobacterium Fortuitum
Authors: da Silva Neto João Pinto , Machado Kyra Nhayanna Coutinho , Roisman Luiz
Published: October 2019

Background

To report a case of a chronic steroid user male patient who developed local abscesses caused by M. fortuitum and concomitant asymptomatic choroidal granuloma.

Case presentation

A 37-year-old african-american male with history of use of anabolic drugs and intramuscular mineral oil injections in the upper and lower limbs for 15 years for muscular hypertrophy. He developed intramuscular abscesses with systemic infection, sub-retinal lesions in both eyes and alterations in cerebrospinal fluid suggestive of mycobacteria. Considering these findings, empirical treatment for tuberculosis was started, without success. After several negative cultures of the material drained from the abscesses, finally one of the cultures isolated the agent Mycobacterium Fortuitum. Proper treatment for atypical mycobacteria was initiated with clinical and laboratory improvement. After 6 months the sub-retinal lesions regressed.

Conclusions

A typical choroidal granuloma caused by M. Fortuitum is a rare presentation of the infection and our report showed a good outcome with proper treatment.
Research Roundup
Authors: Paterson Ruth
Published: October 2019

Ruth Paterson provides an overview of recently published articles that may be of interest to non-medical prescribers. Should you wish to look at any of the papers in more detail, a full reference is provided
Psychoactive drugs and the microbiome
Authors: Winter George
Published: October 2019

George Winter discusses the question whether microbes in the gut can affect the brain, and if it is possible to treat brain disorders by changing the composition of gut microbiota
The management of hyperglycaemia of inpatients receiving enteral feeding: a practical guide
Authors: Menon Vaishnavi , Kalathil Dhanya
Published: October 2019

Enteral feeding is often associated with hyperglycaemia, regardless of if there has been a previous diagnosis of diabetes mellitus. Hyperglycaemia in hospitalised patients is associated with increased morbidity, mortality and length of hospital stay. Insulin therapy is used to manage EF-related hyperglycaemia. Due to several options and regimens being available, choosing and managing an appropriate insulin regimen for EF is challenging and is best undertaken using a multidisciplinary team approach. Inappropriate or incorrect insulin use can lead to poor glycaemic control and/or hypoglycaemia. This article discusses the various treatment strategies for hyperglycaemia associated with EF, prevention of hypoglycaemia, the role of the MDT in providing optimal care and suggestions for simplifying the management of these patients.
Do you know what contracts you are entering into?
Authors: Odell Kirsty
Published: October 2019

Kirsty Odell, senior solicitor in the corporate healthcare team of law firm Hempsons, looks at potential liability for practices entering contractual arrangements
Lightening the cognitive load for your students
Authors: Bromley Matt
Published: October 2019

Classroom consistency helps pupils to learn. Drawing upon Miller's Law on the capacities of our working memory, Matt Bromley looks at what consistency means in practice
Succeeding with vulnerable learners
Authors: Martindale Darren
Published: October 2019

Supporting vulnerable or looked after students with complex needs or difficulties around behaviour, emotional wellbeing or social development is challenging. Drawing on 20 years of experience, Darren Martindale has five golden steps that can make the difference…
How schools can respond to climate change strikes
Authors: Barton Geoff
Published: October 2019

The strikes in protest at the climate change crisis have made international headlines. Geo? Barton looks at how schools might work with pupils to help them engage with climate change
Supporting children through crisis moments
Authors: Knightsmith Dr Pooky
Published: October 2019

When a young person with mental health or wellbeing issues faces a crisis moment, there are some simple techniques and ideas that could help. Dr Pooky Knightsmith explains
Getting the most out of homework
Authors: Dabell John
Published: October 2019

What does the educational research say about the merits of homework and about how we should be using and setting homework in secondary education? John Dabell takes a look
Governors and CPD
Authors: Cunningham Maria
Published: October 2019

How much does your school's governing board know about effective staff development and training – and what impact and influence should they be having on your CPD strategies? Maria Cunningham offers some guidance
Supporting your staff: 36 ideas for your school
Authors: Radford Peter
Published: October 2019

How can we ensure the teachers in our school are engaged, motivated and committed to remaining in the profession? Peter Radford offers 36 actions school leaders can take
Journalistic griots: The marginalization of indigenous language news and oral epistemologies in Ghana
Authors: Mohammed Wunpini Fatimata
Published: October 2019

This study examines news production and newsroom culture in radio stations in Ghana’s Northern Region. It explores the dynamics of news production and delivery in indigenous language newsrooms. Through in-depth interviews with eight indigenous language news presenters and journalists, the study critically explores the intricacies of news production, drawing attention to how news production is contextualized within this society. Through an oral epistemological approach, I argue that news journalists and presenters draw on orature and oral epistemologies to build their news-presenting personas and personalities in a way that positions them as frame sponsors who intentionally set the agenda for news content by unilaterally selecting specific stories to air. This study presents novel ways to conceptualize framing and agenda-setting while demonstrating the usefulness of customizing theory for specific sociocultural contexts. The study presents theoretical and practical implications to bridge the gap between theory and praxis while rethinking news production in Global South contexts such as Ghana.
Volemic changes associated with heparin administration to patients with thyrotoxicosis before plasmapheresis
Authors: Lukomsky G. I. , Alexeyeva M. Ye. , Godzhello M. A.
Published: October 2019

The time course of plasma volume was followed up in patients with thyroid diseases injected heparin before plasmapheresis which was carried out as a measure of preparation to surgery. A tendency to normalization of circulating plasma volume after heparin injection was detected and statistically confirmed.
Increased contrast magnetic imaging in the diagnosis of hypophyseal adenoma
Authors: Belichenko О. I. , Dedov I. I. , Marova Ye. I. , Zenkova T. S. , Shariya M. A.
Published: October 2019

Magnevist (Schering, Germany) was tried for magnetic imaging using Magnetom-P63 (Siemens) superconducting magnetic resonance system at a magnetic field intensity of 1.5 T. Magnetic imaging of the brain was carried out in 24 patients with hypophyseal adenomas before and after intravenous injection of Magnevist. Use of this agent helped more distinctly differentiate between adenoma tissue and the adjacent parasellar structures of the brain, thus appreciably improving the quality of diagnosis of hypophyseal adenoma by magnetic imaging. Recommendations on the use of Magnevist for examination of the sellar region by magnetic imaging are offered.
Stromal ovarian tecomatosis
Authors: Maltseva M. F. , Pishchulin A. A. , Bronstein M. E.
Published: October 2019

In 1942, S. Geist and J. Geines were the first to publish the unusual results of a histological examination of ovarian tissue in patients with Stein-Leventhal syndrome. They found foci of "luteinized" tecal cells, similar to theca interna folliculi cells, scattered along the ovarian "stroma" outside the follicles. The following year, L. Fraenkel in his work called these histopathological changes in ovarian hypertosis. In 1949, A. Culiner and S. Shipel used the term "hypertension syndrome" to refer to conditions in which foci of "tekomatosis", consisting of 10-30-60 individual hypertrophic and luteinized tecal cells in the ovarian "stroma", combined with the clinical picture of defemini ation and virilization. In subsequent years, due to the lack of a unified terminology in the literature, synonyms such as hypertosis, cortical stromal hyperplasia, stromal proliferation, stromal tecosis, and tecomatosis were used. R. Feinberg proposed dividing hypertosis into 3 groups, based on the predominance of cystic follicles and a thickened white membrane (mixed tecosis) with increased proliferation of the actual cells of the interstitial tissue or without it (internal tecosis). The presence of "stromal" proliferation in the absence of phenomena of cystic atresia of the follicles, he defined the term "stromal tecosis" (tekomatosis). I.V. Golubeva for the first time in our country drew attention to the clinical and hormonal features of stromal tecomatosis (ST), which was previously considered in the framework of polycystic ovary syndrome, and after 3 years M.E. Bronstein proved that this is an independent morphological form and gave a definition concepts of ST. Ovarian CT is understood as hyperplasia of the interstitial tissue of the ovaries and the appearance in it of groups of hypertrophied epithelioid cells that form foci of various sizes without any connection with follicles. The subject of this review is the consideration of the clinical and morphological picture of this disease, the features of hormonal homeostasis, modern hypotheses of the pathogenetic mechanisms of its development, as well as possible treatment options.
No need for surgery? Patterns and outcomes of blunt abdominal trauma
Authors: Goedecke Maximilian , Kühn Florian , Stratos Ioannis , Vasan Robin , Pertschy Annette , Klar Ernst
Published: October 2019

Introduction

The management of a patient suffering from blunt abdominal trauma (BAT) remains a challenge for the emergency physician. Within the last few years, the standard therapy for hemodynamically stable patients with BAT has transitioned to a non-operative approach. The purpose of this study is to evaluate the outcome of patients with BAT and to determine the reasons for failure of non-operative management (NOM).

Materials and methods

Analysis of 176 consecutive patients treated for BAT was conducted in a German level 1 trauma center from 2004 to 2011. Abdominal injuries were classified according to the American Association for the Surgery of Trauma (AAST). Patients included were demonstrated to have objective abdominal trauma with either free fluid on focused assessment with sonography for trauma (FAST) or computed tomography (CT), or proven organ injury.

Results

Patients, 142 of 176 (80.7%), with BAT were initially managed non-operatively, with a success rate of 90%. The rates of NOM success were higher among those with less severe injuries; 100% with Abbreviated Injury Scale (AIS) of 1. In total, 125 patients (71.0%) were managed non-operatively, and 51 (29.0%) required surgical intervention. NOM failure occurred in 9.2% of the patients, the most common reason being initially undiagnosed intestinal perforation (46.2%). Positive correlation was identified (r = 0.512; p < 0.001) between the ISS (injury severity score) and the NACA (National Advisory Committee of Aeronautics) score. The delay in operation in NOM failure was 6 h in patients with underlying hepatic or splenic rupture and 34 h with intestinal perforation. The overall mortality of 5.1% was attributed especially to old age (p = 0.016), high severity of injury (p < 0.001), and greater need for blood transfusion (p < 0.001).

Conclusion

NOM was successful for the vast majority of blunt abdominal trauma patients, especially those with less severe injuries. NOM failure and operative delay were most commonly due to occult hollow viscus injury (HVI), the detection of which was achieved by close clinical observation and abdominal ultrasound in conjunction with monitoring for rising markers of infection and by multidetector computed tomography (MDCT) if additionally indicated. Based on this concept, the delay in operation in patients with NOM failure was short. This study underscores the feasibility and benefit of NOM in BAT.
Crystal structure of (3R,5R,8R,9R,10R,12R,13R,14R)-4,4,8,10,14-pentamethyl-17-((R)-2,6,6-trimethyltetrahydro-2H-pyran-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthrene-3,12-diol, C30H52O3
Authors: Zhao Ruo-Lin , Wang Hui-Yun , Luan Ming-Zhu , Zheng Xia , Zhao Feng-Lan , Meng Qing-Guo
Published: October 2019

C30H52O3, orthorhombic, P212121 (no. 19), a = 6.71147(14) Å, b = 16.6565(4) Å, c = 25.0815(6) Å, V = 2803.85(11) Å3, Z = 4, R
gt (F) = 0.0405, wR
ref (F
2) = 0.1092, T = 293(2) K.
Synthesis and antifungal activities of 3-substituted phthalide derivatives
Authors: Fan Lingling , Luo Bilan , Luo Zhongfu , Zhang Li , Fan Judi , Xue Wei , Tang Lei , Li Yong
Published: October 2019

In order to obtain novel bioactive compounds with significant antifungal activities, two series of 3-substituted phthalide derivatives were designed and synthesized via reduction, bromine substitution, and etherification. In addition, the antifungal activities of all target compounds against nine phytopathogenic fungi in vitro were tested by using the mycelial growth rate method at the concentration of 50 μg mL−1. Preliminary bioassay tests showed that some compounds exhibited more potent antifungal activities as compared with hymexazol. The preliminary structure-activity relationships (SARs) of all target compounds were also investigated.
Prevalence of High-Burden Medical Conditions Among Young and Middle-Aged Adults With Pediatric-Onset Medical Conditions: Findings From US Private and Public Administrative Claims Data
Authors: Whitney Daniel G.
Published: October 2019

Adults with pediatric-onset medical conditions (POMCs) are susceptible to early development of high-burden medical conditions. However, research pertaining to this topic is lacking, which is vital information that could assist in health benefit planning and administration. The purpose of this study was to determine the prevalence of high-burden medical conditions among privately and publicly insured adults with POMCs, as compared to adults without POMCs, from the US. Data from 2016 were extracted from Optum Clinformatics® Data Mart (private insurance) and a random 20% sample from Medicare fee-for-service (public insurance). International Classification of Diseases, Tenth Revision, Clinical Modification (ICD-10-CM) diagnosis codes were used to identify 18–64-year-old beneficiaries with POMCs, as well as several high-burden medical conditions, including pain, fracture, mood affective disorders, anxiety disorders, ischemic heart diseases, cerebrovascular diseases, hypertensive and other cardiovascular diseases, type 2 diabetes, osteoporosis, osteoarthritis, chronic obstructive pulmonary diseases, liver diseases, and cancer. Privately and publicly insured adults with POMCs had higher prevalence of all medical conditions compared to adults without POMCs. Publicly insured adults with POMCs had higher prevalence of all medical conditions compared to privately insured adults with POMCs, except for the lower prevalence of pain and cancer. When stratified by the category of POMCs (eg, musculoskeletal, circulatory), privately and publicly insured groups tended to have higher prevalence of most (private) or all (public) medical conditions compared to adults without POMCs. Adults with POMCs have higher prevalence of several high-burden medical conditions compared to adults without POMCs. This health disparity was present regardless of insurance coverage, but was generally more pronounced for public vs. private insured adults with POMCs.
Participación de Rhipicephalus sanguineus en la transmisión de Anaplasma platys en caninos
Authors: Forlano María Dalila , Uzcategui Juan , Mujica Franklin , Orellana Nelson
Published: October 2019

El objetivo del estudio fue determinar la participación de la garrapata Rhipicephalus sanguineus en la transmisión de Anaplasma platys en caninos. Se inoculó un canino con una cepa de A. platys y una vez que desarrolló parasitemia confirmada por frotis y por PCR, se le infestó con ninfas y adultos de R. sanguineus. Se colectaron las garrapatas ingurgitadas, se colectó el intestino y la glándula salival de las garrapatas alimentadas en el canino inoculado y se realizó un PCR. No se amplificó ADN de A. platys en el macerado de ninfas, en adultos pos-muda sin alimentar, o en adultos alimentados del canino inoculado experimentalmente. Los resultados indican que la R. sanguineus no participa en la transmisión de A. platys.
Eficacia de florfenicol para el tratamiento de infecciones por Pasteurela multocida y Mannheimia haemolitica en alpacas (Vicugna pacos)
Authors: Ferrante Marcos , Rezler Wosiacki Sheila
Published: October 2019

El objetivo del trabajo fue evaluar el efecto del tratamiento con florfenicol en dosis de 20 mg/kg por vía i.m. en el tratamiento de infecciones causadas por Pasteurela multocida y Mannheimia haemolitica en alpacas. Se determinó la tasa de eficacia según la concentración inhibitoria mínima (CIM) de la bacteria infectante. Para esto, fue realizada una simulación de Monte Carlo de los parámetros farmacocinéticos para 10 000 eventos y el análisis farmacocinético/farmacodinámico (PK/PD). Las tasas de eficacia estimadas para P. multocida fueron del 99, 20 y 1% según las CIM de las cepas 0.5, 1 y 2, respectivamente, mientras que las tasas de eficacia estimadas para M. haemolitica fueron de 100, 95 y 3% según las CIM de las cepas 0.5, 1 y 2, respectivamente. La probabilidad de obtener la cura bacteriológica después del tratamiento disminuyó de manera altamente significativa para infecciones causadas por bacterias con CIM superiores a 0.5 µg/ml para P. multocida (p˂0.01) y 1 µg/ml para M. haemolitica (p˂0.01). Los resultados evidencian la necesidad de incorporar en el protocolo terapéutico el aislamiento bacteriológico, la determinación de la CIM y la optimización de la dosis terapéutica según la susceptibilidad de las cepas de P. multocida y M. haemolitica a fin de evitar fallas terapéuticas y, consecuentemente, potenciar el desarrollo de resistencia bacteriana.