bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hepcidin Dysregulation in Patients with Type 1 Gaucher’s Disease.
Authors: Hughes Derralynn A. , Li Anna , Keshav Satish , Mehta Atul 
Published: October 2019

Abstract
Gaucher’s Disease is a glycosphingolipid storage disorder where accumulation of glucosylceramide within reticuloendothelial cells results in diverse systemic manifestations. Patients often present with multifactorial anaemia and elevated ferritin which in the context of chronic inflammation may not truly represent their iron status.
We have investigated the iron status and the role of of hepcidin, a negative regulator of iron absorption, in our cohort of patients with type 1 Gaucher Disease. Analysis of haematological indices and haematinic parameters of 57 patients with type 1 Gaucher disease revealed 31% males and 29% females to be anaemic. Of these haematinic parameters were consistent with iron deficiency or anaemia of chronic disease in 45.5% and 27.3% of patients respectively. Measurement of the soluble transferrin receptor (sTFR), which is elevated in iron deficiency and reduced in iron overload, allowed the iron status of patients to be further classified. Of 17 patients in whom the sTFR was measured 64.7% where found to have normal levels, 11.7% had elevated levels indicative of iron deficiency, and 23.5% reduced levels indicating iron overload. The mean ferritin was 79ug/l and 547ug/l in these groups respectively. Levels of serum prohepcidin, the 84 amino acid precursor of hepcidin, were measured using a competitive elisa (DRG diagnostics). Levels of prohepcidin in those patients with normal sTFR were comparable to levels in control subjects: 387.2 +/− 305ng/ml compared to 357 +/1 120 ng/ml. In those patients with elevated sTFR and iron deficiency the prohepcidin level was appropriately suppressed to 112.5 +/− 17.6 ng/ml. Similarly levels in anaemic Gaucher patients were lower than those in patients with normal haemoglobin. However, in those patients with low sTFR and high ferritin the prohepcidin measuring 225 +/−35.3 ng/ml was not appropriately elevated compared to control subjects or Gaucher patients with normal iron status. We hypothesize that storage of glucosylceramide within lysosomes of macrophages interferes with their iron sensing mechanisms and secretion of regulatory cytokines. Iron overload is not appropriately detected and hepatocyte synthesis of hepcidin not upregulated. Progressive accumulation of iron may therefore occur.
Dynamic Defense System Against HMGB1, a Proinflammatory and Lethal Mediator in Severe Infections and Malignancies.
Authors: Ito Takashi , Abeyama Kazuhiro , Kawahara Ko-ichi , Biswas Kamal K. , Uchimura Tomonori , Yamada Shingo , Hashiguchi Teruto , Maruyama Ikuro 
Published: October 2019

Abstract
High Mobility Group box 1(HMGB1) is an abundant DNA-binding protein that acts as a proinflammatory cytokine when released in the extracellular milieu by necrotic and inflammatory cells. Moreover, an increased HMGB1 in the circulation of septic patients may induce multi-organ failure and lethality. However, very recent observations suggest that the protein also acts as an innate adjuvant, stem cell chemoattractant and growth factor. Thus only systemic and circulatory HMGB1 may induce morbidity and mortality, however, localized HMGB1 may have beneficial effects. Therefore, we serially examined the serum HMGB1 level in patients with various diseases, and also evaluated the significance of the protein.
We demonstrate here how HMGB1 is localized and acts as an immune-adjuvant and a repairing factor in damaged tissue. We first established specific ELISA method to measure HMGB1. An increased level of HMGB1 was detected in the serum from patients with sever sepsis, infections, malignancy and so on. However, serum HMGB1 concentrations were fluctuated during the clinical course, and could not be concluded as a lethal mediator as previously reported. Next we investigated the reason of dynamic fluctuations of the protein in the circulation. Based on our findings, we proposed that this fluctuation of HMGB1 concentrations may be mediated by at least following three fashions; 1) proteolytic degradation by plasmin and thrombin, 2) endothelial thrombomodulin(TM) adsorption, and 3) generation of antibody against the protein. We observed that plasmin efficiently degraded HMGB1 into small fragments. However, interestingly the generated fragments of the protein still possess an ability to produce TNFa in macrophages through an undefined pathway. TM binds the protein on its N-terminus lectin-like domain. Binding of HMGB1 to TM resulted in decrement of TM’s cofactor activity to activate protein C by thrombin. HMGB1 bound to TM was gradually degraded by thrombin. These may be a system to localize HMGB1 only in injury sites where TM is down-regulated or disappeared through endothelial-loss. This may exert endothelial defense system against extracellular HMGB1 in severe tissue injury. Another possibility is that the generated antibody against HMGB1 may neutralize the proinflammatory action of the protein. In this context, we found that some of the antibodies against HMGB1 have the characteristics of P-ANCA(perinuclear anti-neutrophil cytoplasmic antibody). This may alter the phenotype of the underlining diseases. In conclusion, we suggest that HMGB1 is not merely a lethal mediator, but a kind of “testament” mediator of cell necrosis or invasive attacks to dendritic cells.
The Effect of IVIG on Fcγ Receptor-Dependent Monocyte Phagocytosis and FcγR Gene Expression.
Authors: Chan Howard H.W. , Li Wei , Su Brenda B. , Denomme Gregory A. 
Published: October 2019

Abstract
BACKGROUND
The mechanism(s) of intravenous immunoglobulin (IVIG) towards inhibition of monocyte phagocytic activity involves the function and/or the expression of inhibitory FcγRIIb in a murine model. To confirm these findings in human monocytes, we used a human monocyte phagocytic model in vitro to study the effects of IVIG on the phagocytic activity and the expression of FcγR genes.
METHODS
Part A: Monolayer Monocyte Phagocytosis Assay
Normal volunteer’s peripheral blood mononuclear cells (PBMC) were isolated from heparin anticoagulated blood by Ficoll-Hypaque (Pharmacia Biotech) density separation. The PBMCs were washed and the monocytes were purified using a magnetic bead-positive selection method with anti-CD14 antibody (Miltenyl Biotec). 105 monocytes were incubated in a microtiter plate at 37°C for 1 hour before exposed to IVIG 0.5 g/L. Anti-D (WinRho) sensitized Rh positive (R2R2) red cells were added to the monocytes at 0.5 hour and 18 hour post-IVIG treatment. After 1 hour incubation with sensitized RBC, monocytes phagocytic activity is measured by chemiluminescence detection with a LumiCount (Packard). The readings were normalized with maximal chemiluminescence signal achieved by the monocytes without prior exposure to IVIG (positive control). Part B: RT-PCR of FcγRIIa and FcγRIIb
After 18 hours of exposure to two different concentrations of IVIG (0.5 and 5 gm/L), monocytes were collected and total RNA was isolated with TRIzol reagent (Invitrogen). 1 μg of RNA was used to generate first strand cDNA using Superscript II RT kit (Invitrogen). FcγRIIa and IIb were amplified with AmpliTaq Gold DNA polymerase system (Applied Biosystems). The PCR products were evaluated by polyacrylamide gel electrophoriesis.
RESULTS
Part A:
Dose-response curves were generated by plotting normalized chemiluminescence against the concentration of anti-D used to sensitize the red cells. Anti-D sensitized red cells were phagocytosed by monocytes in a dose-dependent manner. There is a time-dependent inhibition of monocyte phagocytosis when monocytes were incubated with IVIG at 0.5 gm/L. (Fig. 1)
Part B:
There is no significant difference in the gene expression of FcRγIIb and FcγRIIa in the adherent monocytes after incubating with either low dose (0.5 gm/L) or high dose (5 gm/L) of IVIG for 18 hours. (Fig 2)
CONCLUSION
Delayed inhibition of phagocytic activity with18-hour exposure to IVIG is not directly mediated via the modulation of FcγRIIb gene expression in human monocytes. Other mechanisms, such as intracellular signalling or receptor coupling, might be involved in the delayed inhibitory effects of IVIG.
Figure Figure Figure Figure
Biological Characteristics of T Cells from CD4 Idiopathic Lymphocytopenia Patients Activated and Expanded Using Xcellerate™ Technology.
Authors: Lawrence Joy , Bonyhadi Mark L. , Cunningham-Rundles Charlotte , Berenson Ronald J. , Long Alice 
Published: October 2019

Abstract
Idiopathic CD4 lymphocytopenia (ICL) is characterized by reduced CD4+ T cell counts in the absence of HIV infection or other defined causes. Patients with ICL generally have CD4+ T cell counts < 100/μL and are at risk for infections. Administration of IL-2 has been reported to reduce the risk of infections in some patients, but CD4+ T cell counts remain low. Currently, there is no available therapy to increase the number of CD4+ T cells in these patients.
We have developed the Xcellerate™ Technology, in which T cells are activated and expanded ex vivo from peripheral blood mononuclear cells (PBMC) using microscopic paramagnetic beads conjugated with anti-CD3 and anti-CD28 monoclonal antibodies (Xcyte™Dynabeads®). In patients with low CD4 counts due to HIV or cancer chemotherapy, administration of T cells activated and expanded using the Xcellerate Technology or similar process leads to significant increases in CD4+ T cell counts to normal levels that are sustained over several months.
In the current study, we tested the ability of the Xcellerate Technology to expand T cells from patients with ICL. We collected data on T cell phenotype, cell expansion, activation marker expression, cytokine secretion, T cell receptor (TCR) repertoire diversity, functional potential and response to restimulation. T cells expanded a median of 902 fold (range 281–1529; n=3 patients), and demonstrated typical induction of surface-activation marker expression, including upregulation of CD25 (IL-2R) and CD154 (CD40L). Phenotypic compositions of the ICL donor PBMC were heterogenous both before and after T cell expansion. Two patients had very high levels of CD4−CD8− T cells, and one of these two patients also had very high levels of γδ+ T cells. Each of these populations was maintained throughout expansion. The other patient displayed high levels of circulating CD3+CD56+ NKT cells that did not proliferate as robustly as other T cells during the expansion process. For two patients, flow cytometric analysis revealed a normal pattern of TCR Vβ surface expression while the remaining tissue had a skewed TCR Vβ distribution pattern. Following initial expansion and subsequent restimulation, T cells rapidly re-expressed surface activation markers and cytokines. In these preliminary studies, T cells from patients with ICL were expanded successfully using the Xcellerate Technology, suggesting that this approach might be used for clinical application in this disease.
A New Class of Chemokine Analogs as Useful Research Tools to Study Chemokine Receptor Function and Promising Therapeutic Agents.
Authors: Huang Ziwei , Kumar Santosh , Choi Won-Tak , Madani Navid , Dong Chang-Zhi , Liu Dongxiang , Wang Jun , An Jing , Sodroski Joseph G. 
Published: October 2019

Abstract
Chemokine receptors play important roles in many physiological processes and are implicated in a wide range of human diseases including acute respiratory distress syndrome, allergic asthma, psoriasis, arthritis, multiple sclerosis, cancer, atherosclerosis and most notably AIDS. To enable the applications of chemokine ligands as probes of receptor biology and pharmacology, and inhibitors of diseases mediated by chemokine receptors, a major problem with the lack of receptor selectivity of these natural chemokines must be overcome. In this study, we have developed a chemical approach combining total protein synthesis and modular modifications to generate a new family of unnatural chemokines termed SMM-chemokines (which refer to synthetically and modularly modified chemokines) with designed receptor selectivity and affinity. A proof of the concept has been provided by applying this strategy to transform a very nonselective chemokine vMIP-II into new analogs with enhanced selectivity and potency for CXCR4 or CCR5, two principal coreceptors for HIV-1 entry. Such novel molecules have been shown subsequently to be valuable probes in gaining insights into receptor binding and signaling mechanisms, and as potent inhibitors to prevent HIV-1 entry and infection. These results strongly support the design concept of these SMM-chemokines and suggest that general applicability of this approach for studying and controlling other chemokine receptors and the diseases they mediate might be anticipated.
Chemical Agonists and High Shear Stress Induce Apoptosis in Human Platelets.
Authors: Leytin Valery , Mykhaylov Sergiy , Allen David J. , Miz Lukasz , Lyubimov Elena V. , Garvey Marie B. , Freedman John J. 
Published: October 2019

Abstract
Apoptosis, or programmed cell death, is appreciated as the main physiologic mechanism that regulates cell life-span and serves for controlled deletion of unwanted cells. Since its discovery in 1972, apoptosis was long attributed exclusively to nucleate cells. It took more than 20 years to recognize apoptosis in enucleated cells cytoplasts and anucleate platelets. During the following years, apoptosis has been demonstrated in platelets treated with natural and artificial agonists, in platelet concentrates aged during storage under standard blood banking conditions, and in animal models of suppressed thrombopoiesis and thrombocytopenia. Other studies documented that mechanical forces (shear stresses) stimulate platelet activation and signaling in the absence of exogenous chemical stimuli. We analysed whether shear stresses can trigger platelet apoptosis, a question that has not yet been studied. Using a cone-and-plate viscometer (CAP-2000, Brookfield Engineering Labs, Inc., Middleboro, MA), we exposed human platelet-rich plasma to different shear stresses, ranging from physiologic arterial and arterioles levels (10–44 dynes/cm2) to pathologic high levels (117–388 dynes/cm2) occurring in stenosed coronary, peripheral or cerebral arteries. We found that pathologic shear stresses induce not only platelet activation (P-selectin upregulation and GPIb-alpha downregulation) but also trigger apoptosis events, including mitochondrial transmembrane potential depolarization, caspase 3 activation, phosphatidylserine exposure, and platelet shrinkage and fragmentation into microparticles, whereas physiologic shear stresses are not effective. Platelets subjected to pathologic shear stresses are characterized by impaired platelet function as shown by the absence of ADP-induced platelet aggregation. Apoptosis changes were also induced by the treatment of platelets with calcium ionophore A23187 (10 μM) and thrombin (1 U/mL). Thus, in the present work, we have demonstrated that platelet apoptosis can be induced by chemical stimuli and by mechanical rheological forces (pathologic high shear stresses). Most of shear-induced activation and apoptosis events occur inside of the platelet, including translocation of CD62 from alpha-granules to the platelet surface, depolarization of mitochondrial inner membrane potential, activation of cytosolic enzyme caspase 3, and translocation of phosphatidylserine from the inner to the outer plasma membrane leaflet. These data suggest that the effects of shear stress on platelet activation and apoptosis are mediated by mechanoreceptor(s) that transmit activation and apoptosis signals to the cell interior. The platelet paradigm of apoptosis induced by chemical agonists and shear stresses suggests that apoptotic cytoplasmic machinery may function without nuclear participation.
Both Platelet ADP Receptors Contribute to Microparticle Formation.
Authors: Kahner Bryan N. , Dorsam Robert T. , Kunapuli Satya P. 
Published: October 2019

Abstract
Microparticles are shed from the platelet membrane upon platelet activation, and they contain negatively charged phospholipids such as phosphatidylserine (PS) on their membrane that bind to coagulation factors and aid in clot formation. While the procoagulant nature of microparticles is clear, the relative contributions of platelet agonists to microparticle formation leading to a thrombus are still unclear. Because secreted ADP from the dense granules has been shown to potentiate granule release, GPIIb/IIIa activation, and platelet thrombin generation, we studied the effect of antagonizing the P2Y1 and P2Y12 receptors on microparticle formation from platelets that are stimulated with convulxin and thrombin. It has been previously shown that a deficiency in the P2Y12 receptor causes an increase in bleeding time, and we found prolonged bleeding times with P2Y1 knockout mice when compared to the wild type mice. Because microparticles are a part of the hemostatic process, we set out to see if there was a contribution of the P2Y receptors on agonist-induced platelet microparticle formation. Flow cytometry was used to measure microparticle formation in human blood in the presence or absence of P2Y1 or P2Y12 receptor antagonists. In addition, we measured agonist-induced microparticle formation in wild type mouse blood and compared the microparticle formation to P2Y1 knockout mouse blood microparticle. Using human blood stimulated with thrombin and convulxin we observed a decrease in microparticle count when administering the P2Y1 antagonist MRS2179 by 17%, and the P2Y12 antagonist AR-C69931MX by 28%. Furthermore, there was a decrease in microparticle formation in P2Y1 knockout mouse blood when compared to the wild type mouse microparticle formation by 13%. These results illustrate that both of the P2Y1 and P2Y12 receptors play an intricate role in microparticle formation in vivo, and may be one more effect underlying the extended bleeding times of patients deficient in the P2Y12 receptor or for the slightly increased bleeding of those taking the P2Y12 receptor antagonist clopidogrel.
Comparison of Clinical Outcomes in Patients with Heparin-Induced Thrombocytopenia Treated with Direct Thrombin Inhibitors.
Authors: Francis John L. , Sandhu Ranjeev , Drexler Alane , Dougherty John 
Published: October 2019

Abstract
Heparin-induced thrombocytopenia (HIT) is a clinical syndrome that has been reported to occur in 3–5% of patients treated with unfractionated heparin. If untreated, 36–50% of patients diagnosed with HIT develop life or limb-threatening thromboses. The occurrence of thrombosis, the most common serious complication of HIT, results in rates of amputation and death of 10–20% and 20–30% respectively. Thus, the desired clinical outcomes in patients with HIT are the prevention of thromboembolic complications, limb amputation, and death. The three direct thrombin inhibitors available in the U.S. are lepirudin, argatroban and bivalirudin. Lepirudin and argatroban have both been shown in clinical trials to significantly decrease the incidence of these complications in patients with HIT. Our institution also has experience with bivalirudin as treatment for HIT. We therefore sought to confirm whether all-cause mortality, length of stay, bleeding rate, time to platelet recovery, absolute change in platelet count following therapy, and percentage of therapeutic APTTs differed among patients treated with these agents at our institution. Data were collected by retrospective chart reviews, and from the Florida Hospital pharmacy computer system. Length of stay was calculated as the time to hospital discharge following the finding of a positive heparin-platelet factor 4 antibody test. For the purposes of comparison, the therapeutic range for APTT was taken as 50–90 seconds. As shown in the table, there were no statistically significant differences in any of the endpoints, when the efficacy and safety of the three direct thrombin inhibitors were compared. It therefore appears that within our institution, each of the direct thrombin inhibitors are equally efficacious and safe in treating the clinical syndrome of HIT, with similar outcomes with respect to length of stay, recovery of platelet counts, incidence of bleeding, and overall mortality. We conclude that selection of a direct thrombin inhibitor can be guided by the patient’s clinical status and organ function instead of efficacy and safety considerations.
Drug n All-cause mortality (%) Length of stay (days) Bleeding (%) Time to platelet recovery (days) Therapeutic APTT (%) Lepirudin 7 28.5 11.0 28.5 5.3 70 Argatroban 20 30.0 15.1 20.0 5.2 65 Bivalirudin 24 25.0 18.7 25.0 5.4 74 P-value NS NS NS NS NS
Treatment of New Cases of Acute Promyelocytic Leukaemia by Arsenic Trioxide.
Authors: Ghavamzadeh Ardeshir , Alimoghaddam Kamran , Ghaffari Hamidolah , Rostami Shahrano , Mortazavi Yousef , Mousavi Asadolah , Tootonchi Mehrangiz , Jahani Mohamad , Hosseini Roholah , Iravani Masoud , Bahar Babak , Aghdami Nasser 
Published: October 2019

Abstract
Purpose: Arsenic Trioxide approved for treatment of relapsed or refractory APL to ATRA. We studied the effects of Arsenic Trioxide as first line treatment of new cases of APL and their follow up.
Material and methods: we studied 73 new cases of APL diagnosed by morphologic criteria and confirmed by cytogenetic, RT-PCR for PML/RARA and/or FISH and followed patients for MRD by sensitive nested RT-PCR.
Our patients were 30 males and 43 females with median age 30+/− 12. Patients treated by infusion of 0.15mg/kg/d of Arsenic Trioxide to complete remission by morphologic criteria or till day +60. In patients who complete remission achieved, after 28 days rest, again we began Arsenic Trioxide 0.15mg/kg/d for 28 days as consolidation.
Results: complete remission were achieved in 66 patients( 90.4%) and 7 early mortality. Median time to complete remission was 30+/−6.4 days. Most common cause of mortality was APL maturation syndrome ( 4 cases)
Most common toxicities during induction phase were, APL maturation syndrome (10 cases), serositis(6 cases) and hepatotoxicity (19 cases).
63 cases(86.3%) are alive with a median follow up of 17+/−12.65 months. 14 relapses observed in our patients and complete remission achieved with re-treatment by Arsenic trioxide in 11 of them. Also we could control 3 fatal bleeding by infusion of activated factor 7(NovosevenÒ) which stopped hemorrhage .
One year and two/three years survival of patients were 86% and 84%. Most common cause of death was APL maturation syndrome in 4 patients and relapse in 3 cases.Conclusion: Arsenic Trioxide is acceptable as first line treatment of APL and its result is comparable to ATRA with chemotherapy.