bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of Iron Load of the Liver and Gum of Patients with Thalassemia Major.
Authors: Caliskan Umran , Canatan Duran , Ciris Metin , Bozkurt Yesim , Caglayan Gurhan , Kuran Mine , Kosaci Sevgi , Oguz Nurgul , Karadogan Cavidan , Balta Nihal , Cosan Ruya 
Published: October 2019

Abstract
Various researchs have being conducted to replace liver biopsy which is accepted as a gold standart to evaluate prognosis and as the best indicator of total body iron storage in thalassemia major cases. Especially liver and heart iron storages determined by MR have been reported to correlate with tissue iron leves. However, these techniques have not been investigated sufficiently. We haven’t encountered any study comparing histopathological evaluation of gum and liver iron storage as an indicator of body iron storage. Therefore we have aimed to investigate and compare gum and liver iron storages by histological method which is techniqually easier and lesser noninvasive method. For this purpose, totally 22 beta thalassemia major, aged beetween 5–18 years, were investigated. As control group, 20 healthy person who experienced a tooth extraction, aged between 5–18 years, were included to study. Both gum and liver biopsies of the patients and only gum biopsies of controls were performed. Biopsy samples were stained with Prussia blue, hematoxyleneosin and Masson trichrome dyes. Collagen type-I and type-II were evaluated by immunohistochemical staining. Results; in all thalassemics, siderosis, fibrosis and hepatitis were found in hepatocellular, kupffer and portal areas. Iron in arteriolar wall and endotelium was seen in 22,7% and 50% of cases, respectively. Collagen type-I was found in 54,5% of cases. A statistically significant correlation was found between liver fibrosis, siderosis and iron overload but there was no significant correlation between liver and gum as fibrosis and siderosis in thalassemics. As a result, although periodontological and gingival problems in thalassemics were significantly important to the controls as clinically and histopatologically, however, there was no significant correlation between gum siderosis and hepatocellular siderosis as an indicator of body iron storage in thalassemic patients.
Pulmonary Dysfunction in Thalassemia Major Patients.
Authors: Gennari Alessandra , Bellagamba Maria Paola , di Stasio Alessandra , Ferraro Andrea , Cogo Annaluisa , Borgna-Pignatti Caterina , Capra Lucetta 
Published: October 2019

Abstract
The improved survival of patients with Thalassemia Major (TM) has allowed new complications to develop in the adult patients. These previously unknown clinical entities require a new approach to the management of thalassemia. Pulmonary dysfunction is one of the least understood, previously unrecognized, complications of TM.
Patients and Methods The aim of our study was to define the spirometric pattern and lung damage in 35 TM patients (18 M and 17 F, age range from 18 to 44 years). All were regularly transfused (pre transfusion hemoglobin level 9+/−0,4 g/dl) and chelated (25 with deferoxamine,5 with deferiprone and 5 with both chelators). Their serum ferritin level was 1071+/−628 ng/ml All patients had normal cardiac ejection fraction. Nine patients were active smokers (10+2 pack/years) while 9 had been smokers in the past (6+2 pack/years). Four had a mild allergic asthma under good clinical control, 1 had a mild pulmonary hypertension and 1 had mediastinal hematopoietic tissue. Pulmonary function tests were performed before transfusion and included pulmonary volumes and flows recorded by spirometry; residual volume (RV) and total lung capacity (TLC)measured by helium closed-circuit rebreathing technique; carbon monoxide diffusion capacity (TLCO)measured by single breath method.
Results: Only the patient with mediastinal masses showed an obstructive pattern with the FEV1/FVC ratio of 53% of predicted value. Thirteen patients (37%)had a restrictive pattern (TLC<80%pred). No correlation was found with sex, age, smoking, asthma diagnosis, ferritin levels, chelation regimen. A decreased TLCO (<70%pred) that can be indicative of parenchymal disease was present in 29 patients (85,8%) even after correction for hemoglobin levels (Hb value equal to 12 g requires adjust of 8%, Hb value equal to 10 g requires adjust of 18%, Hb value equal to 7 g requires adjust of 45%). The restrictive lung function abnormality with reduced diffusion capacity found in our patients is suggestive of interstitial lung disease secondary to TM. We were unable to correlate the restrictive and interstitial lung disease to iron loading nor to chelation strategies. Further studies are needed to better understand these results and to determine whether different chelation regimens have different impact on pulmonary function. In order to minimize lung damage, however, every effort should be made to educate TM patients not to smoke, and to identify and properly treat TM patients with asthma.
Proteomic Detection of Beta Thalassaemia Trait Using Electrospray Mass Spectrometry-Mass Spectrometry.
Authors: Daniel Yvonne , Turner Charles , Dalton R. Neil 
Published: October 2019

Abstract
Introduction Mass spectrometry has proved a valuable tool in the unequivocal characterisation of haemoglobin variants detected during routine haemoglobinopathy screening using High Performance Liquid Chromatography (HPLC) and/or isoelectric focusing. We have recently described a rapid and specific electrospray mass spectrometry-mass spectrometry (MSMS) method, using proteomic mutation targeting, for simultaneous detection of the clinically significant haemoglobinopathies; haemoglobin (Hb)S, HbC, HbE, HbDPunjab and HbOArab. Here we report additional transitions within the 1min cycle time, using delta chains as a surrogate HbA2, with the objective of detecting beta thalassaemia trait.
Method Blood samples are digested using trypsin, which results in specific cleavage of globin chains at peptide bonds adjacent to lysine and arginine residues, to produce a series of well-characterised peptides. There is strong amino acid sequence homology between the beta and delta chains of haemoglobin, however tryptic digestion produces series of 15 (T1–T15) and 16 (T1–T16) peptides, for beta and delta chains respectively, with differences in T2, T3, T5, T10, T12, T13 and T14. In this report we have investigated the diagnostic utility of T2: beta sequence SAVTALWGK, average mass 932.1 daltons, delta sequence TAVNALWGK, average mass 959.1 daltons. In multiple reaction monitoring mode the respective T2 [M+2H]2+ ions, m/z 466.8 and 480.3, are selected, fragmented, and the singly charged ions of the informative peptide fragments, VTALWGK and VNALWGK, detected.
Results The percentage area ratio of delta/(beta+delta) was calculated and compared with the classical percentage HbA2 value obtained by HPLC. 66 samples with HbA2 values within our normal range (mean 2.7%, range 1.8–3.4), and 58 with HbA2 values indicative of beta thalassaemia trait (5.2%, 4.2–7.9) were analysed by MSMS and the corresponding delta/(beta+delta) ratios were 1.7%, 0.9–2.3 and 3.4%, 2.5–6.0. In addition, in 2 further HbS beta thalassaemia zero compound heterozygote samples confirmed by beta gene sequencing the ratios were 4.1% and 3.7%.
Conclusion This relatively small study provides evidence that beta thalassaemia trait may be diagnosed by MSMS and represents another step towards comprehensive MSMS population screening for clinically significant haemoglobinopathies.
Myocardial T2* in Patients with Cardiac Failure Secondary to Iron Overload.
Authors: Tanner Mark A. , Porter John B. , Westwood Mark A. , Nair Sunil V. , Anderson Lisa J. , Walker J. M. , Pennell Dudley J. 
Published: October 2019

Abstract
Introduction: Myocardial iron overload is a well established cause of heart failure in a number of haematological disorders and is the leading cause of death in β-thalassaemia major (TM). Once overt heart failure develops prognosis is very poor and it would therefore be desirable to identify patients at risk, prior to the development of symptomatic heart failure. Myocardial iron can now be rapidly and reproducibly assessed using a validated cardiac magnetic resonance (CMR) T2* technique. Left ventricular ejection fraction has been demonstrated to relate to myocardial T2* (normal >20ms), and accordingly iron overloaded patients with symptomatic heart failure are likely to have a low T2* but there are no reports of the myocardial T2* level in newly presenting patients with heart failure and the threshold for increased risk is also unknown.
Purpose: To establish the distribution of myocardial T2* values in patients presenting with symptomatic heart failure secondary to iron overload.
Methods: Database records of CMR T2* assessments over a 5 year period were reviewed to identify iron overloaded patients presenting with heart failure.
Results: 28 patients (median age 29y, 11–79) with iron overload and documented heart failure were identified. 22 patients had thalassaemia major, 3 hereditary haemochromatosis, and 3 had miscellaneous transfusion dependent anaemias. The mean myocardial T2* in all groups was 6.8+/− 2.2ms reflecting severe iron loading by clinical criteria. See figure 1. Myocardial T2* values were similar between sub-groups as follows: TM patients 6.7+/− 2.4ms, haemochromatosis 7.7ms (6.7–7.4ms), miscellaneous 6.8ms (4.8–9.1ms).
Conclusion: In this database review, the patients developing heart failure secondary to iron overload all had abnormal myocardial T2*, and overall 89% of heart failure patients had values less than 10ms, which reflects severe myocardial iron loading. This data suggests a myocardial T2* <10ms should be considered a threshold for risk of heart failure (with its attendant high mortality) and such patients should be treated aggressively with increased iron chelation. It also suggests that clinical heart failure occurs nearly exclusively in patients with severe myocardial iron loading.
Distribution of myocardial T2+ values in symptomatic heart failure (n=28) Distribution of myocardial T2+ values in symptomatic heart failure (n=28)
Reduced Intensity Conditioning Matched Unrelated Donor Allograft for Kostmann Syndrome.
Authors: Thachil Jecko , Pizer Barry , Caswell Mark , Keenan Russell 
Published: October 2019

Abstract
Kostmann syndrome is a congenital disorder of myelopoeisis characterized by an absolute neutropaenia and severe bacterial infections. The introduction of G-CSF has dramatically changed the prognosis of these patients in recent years; though in G-CSF refractory patients, haemopoeitic stem cell transplant is still the only effective treatment. We describe the first case report of a patient who, unresponsive to escalating doses of G-CSF, underwent reduced intensity conditioning, Matched Unrelated Donor (MUD) allograft and has been cured from this condition. A male child presented shortly after birth with infection of the umbilicus. He developed repeated infections in the first six weeks of his life including pneumonia and scrotal abscess. Investigations at this stage included a bone marrow examination, which confirmed a diagnosis of severe congenital neutropaenia and he was started on treatment with G-CSF at 5μg/kg/day. Since diagnosis, he had numerous episodes of ear and skin infections and recurrent abscesses, most often with Methicillin resistant Staphylococcus aureus (MRSA) that required repeated hospitalisations. As the patient did not have a matched related donor, dose escalations were tried with G-CSF at 14-day intervals though he did not respond to doses up to 160μg/kg/day. At this stage, we had a child who was not responding to G-CSF injections, had several infections and was a carrier of MRSA and also Vancomycin Resistant Enterococci, who did not have a matched sibling donor. It was decided then to proceed with a MUD transplant with a non-myeloablative prepatory regimen. The pre transplant conditioning included fludarabine 30mg/m2 from day −8 to day −5, CAMPATH-1H 0.2 mg/kg from day -6 to day −2 and thiotepa 250 mg/m2 from day −4 to −2. Ciclosporin was started from day −1 and GCSF (Lenograstim) at 5μg/kg from day +7. A total of 10.4×106 CD 34+ cells/kg recipient’s weight of stem cells was infused. The patient successfully engrafted with a neutrophil count greater than 0.5×109/L on day 12 with an unsupported platelet count reaching above 50×109/L on day 33. The post transplant period was complicated with grade II skin and gut GVH that responded well to increased doses of ciclosporin alone. Slight falls in neutrophil count were noted on days 28 and 113 after transplant, which resolved by adjusting ciclosporin dosage. He is now ten months post transplant and has not had any infections needing hospitalization and maintains his neutrophil count.
Conclusion
We have therefore demonstrated for the first time that reduced intensity conditioning transplant from a matched unrelated donor is a safe and effective treatment for uncomplicated Kostmann syndrome.
Molecular Characterization of the Cannabinoid-Mediated Migration of Monocytes.
Authors: Park In-Woo , Anand Appakkudal R. , Groopman Jerome E. 
Published: October 2019

Abstract
2-Arachidonoylglycerol (2-AG), an endogenous ligand for the cannabinoid receptors CB1 and CB2, functions as a chemokine for monocyte migration. However, the molecular mechanism of its chemotactic effects is not clear. We found, consistent with previous data, that 2-AG induces the migration of differentiated but not undifferentiated monocytic cells in a dose-dependent manner. We first asked whether the expression of cannabinoid receptors changed during monocytic differentiation. Treatment with 1,25-(OH)2vitamin D3, a potent inducer of monocyte differentiation, or with 2-AG did not alter the surface expression of the CB1 and CB2 receptors, indicating that signaling downstream of receptor ligation accounted for the observed effect on monocyte migration. In addition, treatment of differentiated monocytic cells with inhibitors for adenyl cyclase and rho kinase blocked the 2-AG-mediated migration, directly implicating these signaling molecules in monocyte motility. Upon eliminating the concentration gradient of 2-AG, the motility of the cells from the upper to lower compartment was sharply reduced, but not completely abrogated. This suggested that the chemotactic effect may not fully explain the observed change in cell migration. Of note, treatment of monocytes with 2-AG resulted in an enhanced secretion of the chemokines MCP-1 and IL-8. Moreover, exposure of the cells to 2-AG inhibited their migration towards MCP-1, while exposure to MCP-1 did not alter migration toward 2-AG. Taken together, our findings demonstrate that intracellular signaling cascades as well as induction of chemokine secretion contribute to the cannabinoid-mediated migration of monocytes.
Protein-Biochemical Characterisation of Wilate®: Determination of the Upper Residual Moisture Limit during Terminal Dry-Heat Treatment (PermaHeat).
Authors: Kannicht Christoph , Stadler Monika , Neisser-Svae Andrea , Gruber Gerhard 
Published: October 2019

Abstract
Purpose: Wilate® is a novel, human plasma-derived, high purity, double virus inactivated coagulation factor concentrate. The safe and effective treatment of von Willebrand disease patients and haemophilia A has been demonstrated in clinical trials. The very high virus safety is achieved by an optimised solvent/detergent (S/D) method and terminal dry-heat treatment (PermaHeat) of the lyophilised product, not affecting the integrity of the proteins, which was investigated in this study.
Methods: Analyses of PermaHeat treated (+100°C for 120 min) Wilate® samples with different residual moisture (RM) contents were performed in order to evaluate the upper RM limit in lyophilised samples. Samples with RM up to 7.3% were subjected to SDS-PAGE, immunoblotting and peptide mapping. Further investigations were based on FVIII:C activity testing and phospholipid binding properties, VWF multimer analysis, Ristocetin cofactor (VWF:RCo) activity and collagen binding (VWF:CB).
Summary of results: The analyses confirmed that the structural and functional integrities of both VWF and FVIII were maintained during PermaHeat of Wilate® at +100°C for 120 minutes when the RM was kept at or below 1.8% during the heating procedure. Changes of protein-biochemical properties were detectable only for RM values exceeding 2%. Denaturation of samples occurred at RM of more than 3.5%.
Conclusion: Beside S/D treatment, optimised PermaHeat treatment of the lyophilised VWF/FVIII product ensures the high viral safety margin of Wilate®. According to the results, the RM content during PermaHeat treatment was specified with a great safety margin to a maximum of 1.6%. This limit is carefully controlled in each vial before heating by RM testing utilising near infrared (NIR). At or below this RM no structural or functional changes of VWF and FVIII were detected after PermaHeat treatment.
Undetected Factor VIII Level in a Type III von Willebrand Disease (vWD) Patient Masquerading as Severe Hemophilia A.
Authors: Mullah-Ali Ali M. , Chan Anthony , Pai Mohan K. , Decker Kay , Moffat Karen A. , Walker Irwin 
Published: October 2019

Abstract
Type III vWD is a rare disorder characterized by absent or <0.01u/ml of ristocetin cofactor (vW:RCoF), and a Factor (F) VIII:C of 0.03–0.10 u/ml. A total absence of FVIII:C has never been reported in type III vWD. We wish to report one such case where there was no measurable level of FVIII:C in a child leading to an initial diagnosis of severe haemophilia A. This patient presented at birth with post circumcision bleeding that was controlled with local measures. A packed RBC transfusion of 12 ml/kg body weight was given to correct acute blood loss. Coagulation studies performed before red cell transfusion revealed a FVIII:C of <0.01u/ml leading to diagnosis of severe haemophilia A. Patient required no further treatment to control bleeding. At 14 months of age the patient presented with an intracranial hemorrhage (ICH). rAHF (KogenateFS®) in doses of 50 to 400 u/kg resulted in poor recovery, half-life of 88 minutes and an in vivo survival of less than 4 hours. There was no FVIII:C inhibitor but vWF:RCoF, vWF:Ag and vWF multimers were totally absent. vWF concentrate (Humate-P®) in doses of 50–100/u/kg as vW:RCoF resulted in good clinical response, appropriate in vivo recovery and survival as shown in graphs below. This patient has made an excellent recovery and is fully functional 8 weeks after the ICH. Further enquiry into the family history revealed consanguinity with marriage between first cousins. A female cousin is also affected with easy bruising. Both parents have normal FVIII:C and borderline low vWF:RCoF. DNA analysis of the parents, propositus and unaffected siblings is in progress. Our case illustrates the importance of excluding Type III vWD at initial diagnosis even when the FVIII:C is <0.01u/ml instead of assuming it to be a severe haemophilia A.
Figure Figure Figure Figure
Development and Performance of the GTI ADAMTS-13 Activity Assay.
Authors: Pressman Melissa A. , Wuitschick Elizabeth , Chance Suzette C. 
Published: October 2019

Abstract
Thrombotic Thrombocytopenic Purpura (TTP) is a disease characterized by the sudden onset of a classical pentad of symptoms: thrombocytopenia, fever, renal insufficiency, neurologic deficit, and microangiopathic hemolytic anemia. It was recently discovered that ADAMTS-13 is the protease responsible for cleaving von Willebrand Factor; deficiency of ADAMTS-13 activity has been demonstrated in the plasma of TTP patients. The lack of ADAMTS-13 activity results in the accumulation of multimers of vWF in the plasma and ultimately intravascular platelet aggregation resulting in the clinical symptoms associated with TTP. Currently there are no commercially available diagnostic assays for TTP. Several laboratories have developed “in house” assays that measure the activity of the ADAMTS-13 protease. However, these assays are cumbersome, complex, difficult to perform, and there is a long turn around time, as they require long incubations. The results are difficult to interpret and are not easily quantitated. In addition, there is no standardization of results from laboratory to laboratory, mainly because the assays use various technologies. Determining the extent to which the ADAMTS-13 enzyme activity is decreased is believed to be important in distinguishing a patient with TTP from those presenting with similar clinical symptoms. Until ADAMTS-13 assays are more quantitative, there is not a good method for studying patients with severe decreases in ADAMTS-13 levels versus those who have some intermediate but less than normal level of enzyme activity. Furthermore, the lack of standardized results makes the assays less useful in monitoring the effectiveness of treatment of the TTP patients. GTI has developed a rapid, quantitative assay for ADAMTS-13 activity that is based on the cleavage of a substrate which consists of a peptide fragment of von Willebrand Factor and subsequent fluorescent detection. An initial study using plasma (obtained from the Blood Center of Wisconsin) from 5 TTP patients demonstrated good correlation between the GTI assay and the collagen binding assay, thus demonstrating feasibility of the assay. A more extensive evaluation using 40 normal samples and 55 samples from suspected TTP patients was initiated, the results of which will be reported. Assay sensitivity/limit of detection (based on dilution studies) was 3% normal ADAMTS-13 activity. Assay precision using 3 samples (mean values of 96%, 55%, and 9% normal ADAMTS-13 activity) run in duplicate over 8 days was evaluated. Assay imprecision was less than 10% cv for all samples tested.