bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Model of Severe Congenital Neutropenia Reveals Signaling Pathways Mediating Mutant Elastase-Triggered Agranulocytosis.
Authors: Aprikyan Andrew A. , Vaisar Tomas , Makaryan Vahagn , Heinecke Jay 
Published: October 2019

Abstract
Severe congenital neutropenia (SCN; Kostmann’s syndrome or infantile genetic agranulocytosis) defines an inheritable hematopoietic disorder of impaired neutrophil production due to a “maturation arrest” at the promyelocytic stage of differentiation in the bone marrow. SCN patients have recurring severe infections and often develop acute myelogenous leukemia. We and others reported accelerated apoptosis and cell cycle arrest of bone marrow-derived myeloid progenitor cells in SCN patients with autosomal dominant and autosomal recessive inheritance. Heterozygous mutations in the neutrophil elastase (NE) gene encoding a serine protease, are present in a majority of SCN patients, but not in healthy members of the family, thus indicating a key role of mutant NE in pathogenesis of this disorder. To date, there are no animal or cellular models of SCN as both the knock-in of mutant NE as well as the knock-out of normal NE failed to result in neutropenia phenotype in mice. The molecular mechanisms of mutant NE-mediated severe neutropenia remain largely unknown.
We hypothesized that mutations in NE expose the protease to a new range of substrates. To explore this proposal, we established a cellular model of SCN based on tetracycline-regulated expression of mutant NE in human promyelocytic tet-off HL-60 cells that very closely recapitulated the human phenotype. Mutant NE expression resulted in a characteristic block of myeloid differentiation - the cellular hallmark of SCN. Expression of the mutant product was associated with a significant reduction in phosphatidylinosytol-3-kinase and phosphorylated PKB/Akt levels and an imbalance of anti-apoptotic Bcl-2 and pro-apoptotic Bax. These alterations contributed to observed dissipation of mitochondrial membrane potential as determined by FACS analysis, aberrant release of cytochrome C, and accelerated apoptosis. Marked changes in actin cytoskeleton that made the cells more rigid appeared to stem from a reduced level of alpha-actinin and elevated level of Rho GTPase.
Immunoprecipitation of cell lysates with elastase-specific monoclonal antibodies followed by mass spectrometric analysis revealed that NE interacted with histone H2B, one of the key components of the nucleosome core of the chromatin. Interestingly, the expression level of histone H2B was substantially reduced in cells expressing mutant NE, therefore supporting the notion of altered substrate specificity of mutant NE. Thus, these observations provide the first evidence that mutant NE affects specific signaling pathways that lead to alterations in cytoskeleton and chromatin reorganization, subsequent apoptosis, and a block of myeloid differentiation in SCN. This cellular model of SCN should provide an invaluable tool for screening potential therapeutic agents capable of preventing maturation arrest and leukemogenesis in subjects suffering from severe congenital neutropenia.
F8 Gene Mutations and ITT Response in the Italian Cohort of Haemophilia A Patients with Inhibitors.
Authors: Tagariello Giuseppe , Salviato Roberta , Belvini Donata , Radossi Paolo , Zanon Ezio , Gandini Giorgio , Castaman Giancarlo , Sartori Roberto 
Published: October 2019

Abstract
Introduction: Immune tolerance therapy (ITT), consists of administration of daily high doses of factor VIII concentrates by intravenous infusions. It can induce tolerance to the exogenous protein and eradicate the antibody response to FVIII, which still represents the main complication of replacement therapy in haemophilia A (HA) severe cases.
Methods: We have investigated 71 HA patients with inhibitors (sixty-nine severe, one moderate and one mild cases). We screened the patients for the causative mutations in the F8 gene using Long Range PCR for the intron22 inversion, multiplex-PCR for intron1 inversion or conformation sensitive gel electrophoresis (CSGE) followed by DNA sequencing for other mutation types.
Results: Diverse genetic defects were detected in the severe cases, with a predominance of gross mutations: F8 gene inversions, large deletions and nonsense mutations account for 68% of the mutations, whereas in the two non severe patients specific missense mutations were identified. ITT has been attempted in 16 HR patients of this cohort but failed in 5 cases.
Conclusion: We confirmed that the presence of inhibitors correlates well with the presence of null mutations, as reported by Schwaab et al (Thromb Haemost 1995). Several predictive factors for ITT outcome have been so far taken into consideration but no correlation has been made between F8 gene defect and ITT response. In our cohort 2 large deletions, 2 intron22 inversions and 1 nonsense mutations failed to respond. For the remaining 2 large deletions no ITT were attempted so no information are available. Large deletions appear to be a high risk genetic factor both for inhibitor development and for long term inhibitor persistence and ITT unresponsiveness. The identification of these patients at very high risk of inhibitor development and ITT unresponsiveness by mutation analysis should therefore be strongly recommended soon after the diagnosis, even because of the high cost of the therapy.
Concomitant Induction of CMVpp65-Specific CD4+ and CD8+ T Cells Using Dendritic Cells Transfected with mRNA Encoding an Invariant Chain-pp65 Fusion Protein.
Authors: Le Ngocdiep , Dannull Jens , Chao Nelson , Vieweg Johannes 
Published: October 2019

Abstract
Human cytomegalovirus (CMV) disease is increasingly recognized as a major cause of morbidity and mortality in post-stem cell transplant (SCT) recipients due to a lack of cellular immunity. Thus, novel therapies that offer the restoration of cellular immunity in SCT patients are highly desirable for clinical use. Cytotoxic T lymphocyte (CTL) responses against CMV represent a major effector arm of the immune system to control viremia. However, it is well established that the efficient induction and persistence of CTL responses in vivo requires the concomitant induction of antigen-specific CD4+ T helper cells. In this study, we sought to determine whether human dendritic cells (DC) transfected with mRNA encoding an invariant chain-CMVpp65 fusion protein (Ii-pp65) were capable of inducing concomitant CMV-specific CTL and CD4+ responses, thereby constituting a useful strategy for immunotherapy of CMV disease. We show that transfection of DC with Ii-pp65 mRNA leads to enhanced stimulation of CMV-specific CTL in vitro (Figure 1). Furthermore, DC expressing Ii-pp65 are potent inducers of primary CMV-specific CD4+ T cell responses as evidenced by ELISPOT analyses of primed CD4+CD45RA+ T cells (Figure 2). Lastly, efficient routing of Ii-pp65 into the MHC class II presentation pathway is demonstrated by confocal microscopy (data not shown). Based on these preclinical findings, we propose a clinical trial to administer DC, transfected with mRNA encoding a chimeric Ii-pp65, to post-SCT patients. Our primary goal will be to prevent CMV infection and reactivation by inducing strong immune reactivities against CMV.
Figure Figure
Role of HIF as a Growth and Survival Factor in Multiple Myeloma.
Authors: Feinman Rena , Abungu Billy , Barlos Dimitrios 
Published: October 2019

Abstract
Given that hypoxia and insulin-like growth factor-1 (IGF-1) induce hypoxia inducible factor (HIF-1)-dependent vascular endothelial growth factor (VEGF) expression in different cancer models, we began to explore the possibility that HIF-1α may act as a proliferative and survival factor in multiple myeloma cells. In this study, we demonstrate that MM cells express constitutive HIF-1α protein levels and that hypoxia and cobalt chloride, a hypoxia mimetic, increase HIF-1α and VEGF expression in several MM cell lines (ARP-1, MM1.S, RPMI8266). In the presence of IGF-1, maximal induction of HIF-1α expression in MM cells under normoxic conditions occurred after 12 hr of IGF-1 and declined thereafter. In contrast to hypoxia which inhibits the proteasomal degradation of HIF-1α, IGF-1 requires de novo protein synthesis of HIF-1α. Pharmacologic inhibition of PI3K/Akt or MAPK signaling pathways prevented IGF-1 induced HIF-1α and VEGF expression in ARP-1 cells. Since 2-methoxyestradiol (2-ME) has been shown to inhibit MM growth and associated angiogenesis, we examined whether 2-ME would inhibit the accumulation of IGF-1 induced HIF-1α expression. 2-ME markedly reduced constitutive and IGF-1-induced HIF-1α protein levels in ARP-1 cells. To further demonstrate that HIF-1α plays a role in the growth and survival of MM cells, RNA interference was used to inhibit HIF-1α expression. in ARP-1 cells. Nucleofection of ARP-1 cells with small interfering RNA (siRNA) targeting HIF-1α inhibited constitutive HIF-1α as well as IGF-1-induced HIF-1α protein levels. HIF-1β protein levels were unaffected. Consequently, the suppression of HIF-1α inhibited the proliferation of untreated and IGF-1 treated ARP-1 cells. More interestingly, IGF-1 did not fully overcome the growth inhibitory effect of dexamethasone in ARP-1 cells that were nucleofected with HIF-1 siRNA.
Taken together, our results suggest that HIF-1 plays a role in the pathophysiology of MM.
Hemin Prophylaxis for Patients with Frequently Recurring Acute Porphyric Attacks: Experience with Five Cases.
Authors: Griffith Harry S. 
Published: October 2019

Abstract
Background
The acute porphyrias are a group of four clinically indistinguishable diseases characterized by sudden, acute episodes of neurovisceral symptoms that can be life-threatening. Attacks are precipitated by a genetic deficiency in one of the enzymes involved in heme biosynthesis, acting in concert with one or more endogenous or exogenous triggers. The resultant stimulation of hepatic heme results in overproduction of highly reactive heme intermediaries - porphyrins and porphyrin precursors - which accumulate in the liver, blood, and bone marrow. A small percentage of patients have attacks that recur every several weeks, even after all known triggers are eliminated. Although hemin prophylaxis has been recommended for such patients (Anderson K, et al. Recommendations for the Diagnosis and Treatment of the Acute Porphyrias. Annals of Internal Medicine. 2005 Mar; 142(6):439–50), no adequate and well-controlled clinical studies have been conducted.
Methods
We report on five patients, all diagnosed with acute porphyria on the basis of elevated levels of urinary aminolevulinic acid and porphobilinogen (4 with acute intermittent porphyria, 1 with hereditary coproporphyria). All had a history of recurring acute porphyric attacks and received hemin prophylaxis for at least one year in an out-patient setting and were previously treated for a minimum of one year in a hospital setting. The dosage and frequency of prophylactic hemin was based on each patient’s clinical picture, and ranged from a one-day course of 1–4mg/kg/day every 10 to 40 days to a four day-course of 1–4 mg/kg/day every 30 days. The outcome of prophylaxis hemin treatment was determined on the basis of frequency of severe attacks requiring hospitalization and both physician and patient assessment of the severity of symptoms.
Results
Five patients (3 female, 2 male), ranging in age from 25–61 years old suffered from frequently recurring acute porphyric attacks, with symptoms that included abdominal pain (5), psychological disturbances (2), peripheral motor neuropathy (2), and nausea (2). Two patients had a significant reduction in the frequency of their acute attacks and three reported a moderate decrease with prophylactic hemin treatment. All five patients experienced a marked improvement in the reduction of the severity of their attacks. Overall, the occurrence of severe acute attacks requiring hospitalization declined by 66%. Three of the five patients did not require hospitalization over the 12 month period of time. One patient experienced phlebitis during one course of hemin treatment.
Conclusions
Hemin prophylaxis can be an effective disease management strategy in acute porphyria patients with frequently recurring acute attacks.
Association of Mild Anemia and Executive Cognitive Function Impairment in High Functioning Community-Dwelling Older Women.
Authors: Chaves Paulo H.M. , Fried Linda P. , Carlson Michelle 
Published: October 2019

Abstract
Background: Anemia in the elderly is common and may be associated with atypical clinical presentations. Although the impact of severe anemia on cognition has been studied in patients with cancer, less is known about the association of mild anemia on cognitive function in the elderly. Executive cognitive functioning (ECF) refers to the execution of complex activities that involve a set of higher-order cognitive abilities primarily dependent on the frontal lobes. ECF plays a major role in the maintenance of the functional status of older adults. Knowledge about potentially modifiable risk factors for ECF impairment in older adults is limited. The goal of this study was to test the hypothesis that mild anemia is associated with ECF impairment in high functioning, community-dwelling older women.
Methods: Cross-sectional analysis of baseline data from the Women’s Health and Aging Study (WHAS) II, 1994–1996, Baltimore, MD. WHAS II is a population-based study of 436 cognitively intact (Mini-Mental State Exam >24) and highly physically functioning women aged 70–80 years. The analytic sample included 364 subjects with Hb >10 g/dL and the following ECF test components: Trail Making Tests part B (TMTB) - primary outcome - and part A (TMTA), and the difference TMTB-TMTA. Multivariate polytomous logistic regression models were used to model the relationship between performance (based on tertiles) on each ECF test as a function of Hb, while controlling for age, education, cardiovascular disease, and a number of other potential confounders, such as hypertension, diabetes, depressive symptoms, calculated creatinine clearance, forced expiratory volume in the first second, thyroid stimulating hormone, total cholesterol levels, body mass index, walking speed, and prevalent mobility difficulty.
Results: Subjects with mild anemia (Hb 10–12 g/dL) performed worst in all ECF tests; i.e., the percentage of subjects in the best performance tertile for each test was lowest for those with Hb within 10–12 g/dL. Estimates from multivariate models revealed that, as compared to those with Hb≥12 g/dL, those with Hb within 10–12 g/dL were substantially less likely to be in the best performance tertile of the TMTB [odds ratio (OR).23, 95% confidence interval (CI):.06–.86; p=.029], TMTA (OR.27, 95%CI:.07–.98; p=.047), and TMTB-TMTA (OR.30, 95%CI:.10–.89; p=.031), even after comprehensive adjustment.
Conclusion: In this cross-sectional study, mild anemia was an independent risk factor for prevalent ECF impairment in high functioning community-dwelling older women. This raises the hypothesis that correction of mild anemia could potentially provide an opportunity for prevention of cognitive and functional decline in older adults. To test this hypothesis, prospective observational studies and clinical trials are warranted.
Essential Role for Activated Leukocyte Cell Adhesion Molecule in Transendothelial Migration.
Authors: Ofori-Acquah Solomon F. 
Published: October 2019

Abstract
Activated leukocyte cell adhesion molecule (ALCAM/CD166) is a member of the immunoglobulin cell adhesion super family, which has been implicated in diverse physiological and pathophysiological events involving cell migration. Hitherto, ALCAM’s role in inflammation has not been determined. In this study, we show ALCAM is involved in controlling migration of mononuclear leukocytes across the pulmonary endothelium. We demonstrated that ALCAM is localized at intercellular junctions in pulmonary microvascular endothelial cells in vitro and in vivo. ALCAM co-localized with multiple adherens junction molecules including cadherins, catenins and Dlg, as determined by confocal microscopy, and these observations were confirmed by co-immunoprecipitation and co-distribution assays. Treatment of endothelial cultures with EGTA and cytochalasin D translocated ALCAM from intercellular junctions to the cytosol indicating a requirement for homotypic cadherin adhesion and an intact endothelial cytoskeleton for maintaining ALCAM at endothelial cell junctions. Collectively, these data supports the conclusion that ALCAM contributes to the adherens junction complex in endothelial cells. To determine ALCAM’s role in leukocyte-endothelial cell interactions, adult Sprague Dawley rats were intratracheally instilled with macrophage inflammatory protein-1, and this treatment caused acute expression of ALCAM exclusively in newly recruited mononuclear but not polymorphonuclear leukocytes in the alveolar airway. Given that no ALCAM reactivity was observed in peripheral blood leukocytes, we concluded ALCAM is activated as part of the phenotypic switch by mononuclear leukocytes transitioning from circulation to interstitial tissue compartments. To determine the physiological relevance of this finding we examined whether ALCAM was required for transendothelial migration using monocyte chemoattractant protein 1 (MCP-1). MCP-1 dose- and time-dependently increased the number of transmigrated THP-1 monocytes across pulmonary microvascular endothelial monolayers. Recombinant soluble ALCAM dose-dependently reduced the number of transmigrated THP-1 monocytes, whereas in control experiments recombinant soluble vascular endothelial cadherin had no effect on transmigration. This study shows for the first time that ALCAM is located at endothelial cell junctions where it is intimately involved in controlling the number of monocytes that pass through endothelial barriers. ALCAM may therefore play an essential role in the response to inflammation by enhancing recruitment of mononuclear leukocytes by inflamed tissues.
A Study on the Etiology of 912 Anemia Patients in General Hospital.
Authors: Zhao Xiaoying , Zhang Ping , Huang Liansheng 
Published: October 2019

Abstract
Objectives To investigate incidence rate and the etiological factors of anemia in general hospital.
Methods We selected 912 patients with anemia from 4198 inpatients in our hospital from Jan.to Feb.2004,and analyzed their clinical data retrospectively. All inpatients had been detected Hb,RBC,MCV and MCHC. We collected 2ml venous blood in anticoagulant EDTA-k2 tube and measured them with Gen.STM System2 blood cell radiomete in 2 hours. some patients had been punctured bone marrow examination. According to diagnostic standard of anemia(male adult Hb<120g/L, female adult Hb<110g/L, gravida Hb<100g/L) and anemia degree(Mild degree anemia: Hb 90~120 g/L, Moderate degree anemia: Hb 60~90 g/L, Severe degree anemia: Hb30~60 g/L, Profound degree anemia: Hb<30g/L), we classified anemia according to clinical types and cell morphology. according to clinical types, anemia was resulted from primary hematologic disease and secondary anemia. Main causes of secondary anemia were chronic disease, blood loss and chronic systemic disease. According to red blood cell morphology, anemia was classified: microcytic hypochromic anemia (MCV<80fl,MCHC<32%), pure microcytic anemia(MCV<80fl,MCHC32%~36%),normocytic normochromic anemia (MCV80fl~100fl,MCHC32%~36%) and macrocytic anemia( MCV>100fl).
Results The incidence of anemia in inpatients in general hospital was 21.72%. 96.71% of patients were mild-moderate degree anemia and only 13.71% anemia patients showed symptom of anemia such as dizzying,drumming. But only 1.10% of patients had been diagnosed by doctors. 6.36% of patients were caused by hematologic disease and 89.74% of patients coexisted primary disease.53.62% of patient were secondary to chronic disease such as tumor,chronic infection, rheumatic disease. 27.85% of patients were secondary to blood loss. Then 8.00% of patients was caused by chronic systemic disease.
Conclusions Anemia existed in various clinical diseases. Misdiagnosis rate of anemia was high. Appropriate treatment of secondary anemia was beneficial for prognosis of primary diseases. All doctors should pay more attention to anemia in general hospital.
Effects of Zotarolimus (ABT-578), Sirolimus, Paclitaxel and Dexamethasone on Cytokine Production by Human Monocytes In Vitro.
Authors: Li Lanlan , Burke Sandra E. , Mack Matthew M. 
Published: October 2019

Abstract
Introduction: Percutaneous coronary intervention (PCI) and stenting are common therapies used to mitigate symptomatic coronary artery stenosis. These mechanical procedures are invariably associated with local and systemic inflammation, including activation and cytokine release from monocytes/macrophages, which are major stimuli for smooth muscle cell proliferation, migration and restenosis. Drug-eluting stents (DES) may potentially attenuate this inflammatory response, provided that the drug has antiinflammatory potency. This study was conducted to determine the effects of common DES agents on the generation of proinflammatory cytokines by stimulated human monocytes in vitro.
Methods: The production of monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-a (TNF-a) and interleukin-6 (IL-6) by LPS-stimulated monocytes was determined by ELISA after exposure to either zotarolimus (ABT-578), sirolimus, dexamethasone or paclitaxel.
Results: The results showed that zotarolimus (ABT-578) and sirolimus were potent inhibitors of MCP-1 production (IC50 = 2.6 ± 0.2 and 5.9 ± 1.9 nM, respectively). Dexamethasone also inhibited MCP-1 production, but it was less potent (IC50 = 626.9 ± 60.8 nM). TNF-a and IL-6 production were dose-dependently inhibited by dexamethasone (IC50 = 5.5 ± 0.2 and 13.0 ± 3.7 nM, respectively). However, the production of neither cytokine was significantly inhibited by exposure to zotarolimus or sirolimus. Stimulated monocytes were not affected by paclitaxel; cytokine release after exposure to paclitaxel was no different from controls.
Conclusions: Zotarolimus and sirolimus are potent inhibitors of MCP-1 secretion by stimulated monocytes in culture. Dexamethasone is a less potent inhibitor of MCP-1 secretion, but effectively inhibits the release of TNF-a and IL-6. The anti-restenotic effect of these agents may, in part, be due to their potent antiinflammatory properties.