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This article introduces a new probability model based on reflected parameter called the reflected Pareto (RP) distribution. The key properties of the RP model are investigated. A simulation study of the RP model is conducted to evaluate the performances of its estimators. A real-life application is considered to examine the performance of proposed model. The different criteria are discussed numerically as well as graphically to show the flexibility of the RP model. The exponential weighted moving average control charts based on the maximum likelihood and modified maximum likelihood estimators for the shape parameter of the RP distribution are obtained. Detailed simulation results of proposed charts are performed to examine and analyze the performance of these charts with three in-control average run length values and two sample sizes. Finally, the application of the proposed control charts is shown by considering a real-life data set.

AimsAcute graft-versus-host disease (aGVHD) is a leading cause of morbidity and mortality following allogeneic haematopoietic stem cell transplantation (HSCT). The aim of this study was to evaluate the clinical utility of a composite biomarker panel to help identify individuals at risk of developing aGVHD, and to help predict and differentiate between severity of aGVHD following T-cell-depleted allogeneic HSCT.MethodsWe retrospectively analysed our cohort of biopsy confirmed patients with aGVHD, who underwent T-cell-depleted HSCT and matched them with negative controls without any evidence of aGVHD. Post-transplant serum samples on days 0 and 7 and at onset of aGVHD were analysed for elafin, regenerating islet-derived 3-α, soluble tumour necrosis factor receptor-1, soluble interleukin-2 receptor-α and hepatocyte growth factor. Biomarker data were combined as composite panels A–F (table 2) using logistic regression analysis. Receiver operating characteristic analysis was performed to study sensitivity and specificity of the composite panels.ResultsOur composite biomarker panels significantly differentiated between aGVHD and no GVHD patients at time of onset (panel E) and reliably predicted severity of GVHD grades at days 0 and 7 post-transplant (panels B and D). The area under the curve for the composite panel at time of onset was 0.65 with specificity, sensitivity, positive and negative predictive values of 100%, 55.6%, 100% and 78.9%, respectively (p=0.03).ConclusionsThis pilot data support the usefulness of these composite biomarker panels in the prediction of severity and diagnosis of aGVHD in patients undergoing T-cell-depleted reduced intensity allogeneic HSCT.

Background: The use of non-invasive inhaled aerosols for pulmonary drug delivery continues to grow. This is due to the many unique advantages this delivery route offers for the treatment of both local and systemic diseases. The physicochemical properties of the formulated drugs as well as the physiology of the lungs play a key role in both the deposition and absorption of the particles. The airway and the alveolar epithelium are targets for the treatment of respiratory diseases. However, particles have to overcome biological barriers before they reach their target and produce an effect. Methods: In vitro aerosol dispersion performance (i.e. aerodynamic size and aerodynamic size distribution) of inhalable particles is quantified by inertial impaction, as required by regulatory agencies for an investigational pharmaceutical inhalation aerosol formulation to be approved for use in patients as a marketed pharmaceutical product. Using inertial impaction in conjunction with cell cultures of various pulmonary cells in situ as bioimpactors has unique aspects in correlating aerodynamic properties with pulmonary cellular behavior including viability and uptake. These can be as co-culture or in single culture, as 3-D multicellular spheroids or 2-D cellular monolayer using different conditions to grow them, such as air-liquid interface culture (ALI) or in liquid covered culture (LCC). Results: evaluation of the currently available in vitro models and the challenges in developing reliable cellular tools to predict the deposition of inhalable particles in the lungs as a function of aerodynamic particle properties is presented in the manuscript. Conclusion: The mechanistic aerodynamic and biophysical properties of inhaled aerosol particles on the entire respiratory tract at the cellular level based on aerodynamic size and aerodynamic size distribution will be better understood with the development of in vitro methods which are described in this work.

The study by Memarnejadian et al describes the use of an in vivo cytotoxicity assay inmice, whereby the lysis of adoptively transferred allogeneic cells is mediated by donor-specific antibody and independent of cytotoxic T lymphocytes, natural killer or natural or natural killer T cells. Supplemental digital content is available in the text.

ABSTRACTStaphylococcus aureusis a versatile bacterial pathogen that produces T cell-activating toxins known as superantigens (SAgs). Although excessive immune activation by SAgs can induce a dysregulated cytokine storm as a component of what is known as toxic shock syndrome (TSS), the contribution of SAgs to the staphylococcal infection process is not well defined. Here, we evaluated the role of the bacterial superantigen staphylococcal enterotoxin A (SEA) in a bacteremia model using humanized transgenic mice expressing SAg-responsive HLA-DR4 molecules. Infection with S. aureusNewman induced SEA-dependent Vß skewing of T cells and enhanced bacterial survival in the liver compared with infection by seaknockout strain. SEA-induced gamma interferon, interleukin-12, and chemokine responses resulted in increased infiltration of CD11b+Ly6G+neutrophils into the liver, promoting the formation of abscesses that contained large numbers of viable staphylococci. Hepatic abscesses occurred significantly more frequently in S. aureusNewman-infected livers than in livers infected with the Newman seaknockout strain, promoting the survival of S. aureusin vivo. This represents a novel mechanism during infection whereby S. aureusutilizes SAgs to form a specialized niche and manipulate the immune system.

Itraconazole and posaconazole are used in the prevention and treatment of invasive fungal infections. However, the oral bioavailability of both compounds varies widely, and dose–serum concentration relationships are poorly defined for these analytes. The aim of this work was to develop and validate a simple assay that could be implemented in most laboratories for the purpose of therapeutic drug monitoring.

Most respiratory infections, diseases and allergic reactions have varying degrees of inflammation. Inflammation is a natural immunodefensive response to the presence of allergens or foreign particles that come into contact or affect the cells and tissues within the respiratory tract. The three main types of therapeutic drug classes available for anti-inflammatory and anti-allergy effects are corticosteroids, antihistamines and decongestants. Corticosteroid drugs for pulmonary inhalation and/or nasal delivery include beclomethasone dipropionate, budesonide, ciclesonide, fluticasone furoate, fluticasone propionate, mometasone furoate, and triamcinolone acetonide. Antihistamine drugs for nasal delivery include azelastine and olopatadine. Two common decongestants available are oxymetazoline and phenylephrine. Another therapeutic class, the anticholinergic agents, such as ipratropium bromide and tiotropium bromide, are used in pulmonary delivery in the treatment of inflammatory diseases such as asthma and chronic obstructive pulmonary disease. The mast cell stabilizer therapeutic class, cromolyn sodium, can be used to prevent and relieve nasal allergic symptoms. Additionally cromolyn sodium was the first dry powder inhaler product for pulmonary drug delivery several decades ago and currently is on the market as a pressurized metered dose inhaler for pulmonary inhalation delivery. Based on the devices used in pulmonary drug delivery, this route can be subdivided into three categories; namely nebulizers, pressurized metered dose inhalers, and dry powder inhalers. Nasal delivery of anti-inflammatory and antiallergy drugs is most commonly available commercially in an aqueous spray form. This article comprehensively reviews and discusses different kinds of drugs used for anti-inflammatory and anti-allergic effects via the pulmonary and nasal delivery route, as well as their mechanisms of action, marketed products, disease state indications while highlighting drug delivery and therapeutic aspects.

ABSTRACTThere currently are commercial fowlpox virus (FPV)-vectored vaccines for use in chickens, including TROVAC-AIV H5, which expresses the hemagglutinin (HA) antigen of an avian influenza virus and can confer immunity against avian influenza in chickens. Despite the use of recombinant FPV (rFPV) for vaccine delivery, very little is known about the immune responses generated by these viruses in chickens. The present study was designed to investigate host responses to rFPV in vivoand in vitro. In cultured cells infected with TROVAC-AIV H5, there was an early increase in the expression of type I interferons (IFN), Toll-like receptors 3 and 7 (TLR3 and TLR7, respectively), TRIF, and MyD88, which was followed by a decrease in the expression of these genes at later time points. There also was an increase in the expression of interleukin-1ß (IL-1ß), IL-8, and beta-defensin genes at early time points postinfection. In chickens immunized with TROVAC-AIV H5, there was higher expression of IFN-? and IL-10 at day 5 postvaccination in spleen of vaccinated birds than in that of control birds. We further investigated the ability of the vaccine to induce immune responses against the HA antigen and discovered that there was a cell-mediated response elicited in vaccinated chickens against this antigen. The findings of this study demonstrate that FPV-vectored vaccines can elicit a repertoire of responses marked by the early expression of TLRs, type I interferons, and proinflammatory cytokines, as well as cytokines associated with adaptive immune responses. This study provides a platform for designing future generations of rFPV-vectored vaccines.

This study examines the various equilibrium in situsecondary structures of the pharmaceutical heteropolypeptide, KL 4, in the solid state, in solution, and in the monolayer state alone and mixed with dipalmitoylphosphatidylcholine (DPPC) and palmitoyloleoylphosphatidylglycerol (POPG). In situsurface circular dichroism spectroscopy, using a method first reported by Damodaran (Damodaran, S. Anal. Bioanal. Chem.2003, 376, 182−188), of equilibrated KL 4, DPPC/KL 4, POPG/KL 4, and DPPC/POPG/KL 4monolayers at the air−water interface was used to examine the in situtwo-dimensional conformation of KL 4. Gravimetric vapor sorption by solid KL 4was used to analyze the effects of water molecules on the conformation of KL 4when confined as a monolayer at the surface of water. Solid-state KL 4conformation was determined by X-ray powder diffraction (XRPD). The equilibrium interfacial and spreading properties were measured at 25 °C, 37 °C, and 45 °C using the Wilhelmy plate method and Langmuir film balance. Equilibrium phase transition temperatures were measured using differential scanning calorimetry (DSC). It was found that solid-state KL 4, which takes up very little water, exhibits β-sheet and α-helix secondary structures, whereas KL 4in solution appears to exist only as an α-helix. KL 4forms a stable, insoluble monolayer, exhibiting β-sheet and aperiodic structures. These structures provide KL 4, when confined in two-dimensions, the structural flexibility to maximize favorable cationic lysine−water interactions and favorable leucine−leucine hydrophobic and van der Waals interactions; while effectively “shielding” the leucine residues away from water. In DPPC/KL 4monolayers, KL 4retains its native β-sheet and aperiodic structures, consistent with phase separation of DPPC and KL 4in bilayers and monolayers. In POPG/KL 4monolayers, KL 4exhibits an increase in aperiodic secondary structures (loss of β-sheet) to maximize favorable electrostatic interactions, consistent with the observed negative deviations from ideal monolayer mixing.

Invariant natural killer T (iNKT) cells are glycolipid-responsive cells with potent immunomodulatory properties. Although iNKT cells have been implicated in cardiac allograft tolerance, whether in vivo triggering of iNKT cells with Th2-promoting glycolipids offers a therapeutic benefit in heart transplantation remains unexplored.

To become competent killer cells, CD8+T cells requirestimulation through signal transduction pathways associated with the T‐cell receptor, costimulatory molecules such as CD28, and cytokinereceptors such as the interleukin (IL)‐2 receptor. We used wortmanninand LY294002, two inhibitors of phosphatidylinositol 3‐kinase (PI3‐K), to study the role of PI3‐K in mouse cytotoxic T‐lymphocyte (CTL)induction in response to mitogenic anti‐CD3 antibody. Anti‐CD3‐inducedCD8+T‐cell proliferation and CTL development wereinhibited dose dependently by both PI3‐K inhibitors. IL‐2 synthesis byanti‐CD3‐activated CD8+T cells was also diminished byPI3‐K inhibition. PI3‐K inhibition resulted in a modest decrease inanti‐CD3‐induced CD4+T‐cell proliferation but failed toaffect IL‐2 expression by anti‐CD3‐activated CD4+T cells. PI3‐K inhibition during CTL induction resulted in decreased levels ofmRNAs coding for granzyme B, perforin, and Fas ligand. In addition, CTLinduced in the presence of PI3‐K inhibitors failed to conjugatenormally with P815 target cells. Exogenous IL‐2 did not reverse theeffects of PI3‐K inhibition on CD8+T‐cell proliferationand CTL induction. These results support the conclusion that PI3‐Kactivation is involved in T‐cell receptor, CD28, and IL‐2 receptorsignaling of CD8+T cells. PI3‐K is, therefore, animportant component of multiple signal transduction pathways involvedin CTL generation.

Cytotoxic T lymphocyte (CTL) development is regulated closely by an intricate series of signals provided by the T‐cell receptor/CD3 complex, cytokines, and costimulatory ligand/receptor systems. In this study, we have explored the role of interleukin (IL)‐12 and CD28 in mouse CTL development. Activation of T cells with anti‐CD3 monoclonal antibody (mAb) in the presence of anti‐CD86 mAb, which prevents CD28‐CD86 interaction, led to decreased production of type 1 (IL‐2, interferon‐γ) and type 2 (IL‐4, IL‐6, IL‐10) cytokines, as well as diminished expression of granzyme B (Gzm B) and reduced cytotoxic effector function. Cytolytic activity in T‐cell cultures that were activated in the presence of anti‐CD86‐blocking mAb alone or in combination with anti‐CD80 mAb could be restored by the addition of exogenous IL‐12 at initiation of culture. The ability of IL‐12 to substitute for CD28‐costimulatory signaling during CTL development was found to be dependent on the presence of IL‐2 rather than interferon‐γ. IL‐2 is required for IL‐12Rβ2 expression by T cells activated in the presence of anti‐CD86 mAb. Moreover, IL‐12Rβ2 expression by T cells activated in the presence of anti‐CD86 mAb is enhanced by IL‐12. We, therefore, conclude that the ability of IL‐12 to substitute for CD28‐costimulatory signaling during CTL development is a result of the interaction of IL‐12 with IL‐12Rβ2 induced by low levels of IL‐2 synthesized by T cells activated in a CD28‐independent manner.

Coordination polymer of unstable zinc borohydride and pyrazine is prepared and used as an stable, efficient and selective bench top reducing agent for a variety of organic compounds.

Ferric dichromate, polyvinylpyridine supported zinc and ferric dichromates complexes are prepared and used as stable, mild, and efficient oxidizing reagents for the oxidation of different organic compounds. Ferric dichromate complex is more efficient.

Three distinct serologically phenotypic mucins have been described in the hog submaxillary glands; A+, H+, and A-H-(In). Alkaline borohydride reductive cleavage of A+and A-pig submaxillary mucin (A+-PSM and A--PSM) has enabled Carlson to propose structures for the oligosaccharides of these mucins. The structures presented readily explained the A+and H+cross-reactivity of the mucins, but left the structures of the In mucins unexplained. The relationship of H+and In mucins and our previously published serological observations prompted us to determine the structures of the oligosaccharides isolated from inactive mucins secreted by hog submaxillary glands.

Prostate cancer is the most common solid cancer among American men. Although there are various modalities for treatment, including radical prostatectomy among many others, the former is, nevertheless, not without any accompanied complications. Other than the well-known surgical complications such as erectile dysfunction, urinary incontinence, and voiding dysfunction, osteomyelitis of the symphysis pubis is not a well-understood complication, with various hypotheses explaining its pathogenesis. Although osteomyelitis of the pubis symphysis has been reported after endoscopic urological procedures such as transurethral resection of the prostate, it has rarely been reported after robotic surgeries. We hereby report, to the best of our knowledge, the first osteomyelitis of the pubis symphysis, after robotic prostatectomy, in a patient with prostate cancer and no previous radiation therapy.

No relevant conflicts of interest to declare.

Background:Acute Graft-versus-host disease (aGVHD) is a leading cause of morbidity and mortality following allogeneic haematopoietic stem cell transplantation (HSCT). The diagnosis and management of aGVHD relies on clinical criteria and graded according to the number and extent of organ involvement. Clinical diagnosis of aGVHD is one of exclusion and usually requires tissue biopsy to aid diagnosis. The aim of this study was to evaluate the clinical utility of a biomarker panel to help identify individuals at risk of developing aGVHD and its ability to differentiate severity at onset of acute graft-versus-host disease (aGVHD) following T-cell depleted allogeneic HSCT.

Poly(2-vinylpyridine)(I) and poly(4-vinylpyridine)(II) supported chromium peroxide reagents are readily prepared and used as mild, efficient, selective and easy to handle oxidizing agents for oxidation of benzylic and allylic alcohols.

Invasive fungal disease (IFD) is a difficult diagnosis in patients undergoing haematopoietic stem cell transplantation (HSCT) or high dose chemotherapy. For clinical trials the revised EORTC/MSG criteria is a useful diagnostic tool but there are few data on its usefulness in clinical practice. The aim of this study was to evaluate the ‘real world’ incidence and outcome of IFD in patients undergoing HSCT or high dose chemotherapy with expected neutropenia of 10 days or more using all approved EORTC/MSG diagnostic tools.Patients were recruited prospectively between December 2008 and May 2010 and followed for at least 4 months after chemotherapy/HSCT or until death. During admission twice weekly galactomannan (GM) was performed and neutropenic sepsis unresponsive to antimicrobials triggered diagnostic work-up with continuous volume acquisition computed tomography (CT) and tissue diagnosis was obtained where possible. β-D-glucan (BDG) was performed on all positive GM samples (n=139), patients with relevant CT abnormalities but negative GM (n=41), and 79 control samples. The CT scans were reviewed independently by two chest radiologists and all cases were independently verified before assigning EORTC status. All patients had antifungal prophylaxis during the period of neutropenia or continuing immunosuppression (fluconazole for low risk patients, itraconazole suspension or posaconazole for high risk patients, and voriconazole for secondary prophylaxis).Two hundred and three patients were recruited [123 male, 80 female; median age 54y (range 19–73); median follow-up 194 days (range 12–647)]. The underlying diagnoses were: acute leukaemia 62 (myeloid 55, lymphoid 7), chronic leukaemia 4 (myeloid 3, lymphoid 1), MDS/MPD 33, aplastic anaemia 19, lymphoma 37 (non-Hodgkin's 29, Hodgkin's 8), multiple myeloma 45, and others 3. Main treatments received were: HSCT 165 (81%) [allogeneic 94, cord 5, autologous 66], chemotherapy 28 (14%), and immunosuppressive therapy (IST) 8 (4%). The total number of treatments received during study period was 263 (allografts 106, chemotherapy 77, autografts 67, IST 13). The patients were heavily pre-treated; 85% had at least one prior therapy with a median of 5 cycles of therapy/patient (range 1–17). There were 44 (21%) cases of IFD in 40 patients: proven 14 (Aspergillus fumigatus 2, Fusarium spp 1, mould IFD 5, non-candida albican spp 5, Pneumocystis jirovecii 1), probable 30. Invasive pulmonary aspergillosis was the commonest presentation (82%). The treatment-specific incidences were 17%, 12%, 7%, and 4% for allogeneic HSCT, chemotherapy, immunosuppressive therapy, and autologous HSCT respectively. Patients with AML/MDS had the highest risk (27 cases) while multiple myeloma represented the lowest risk (1 case). Median time to diagnosis of IFD was 42 days (range 3 to 524) from index chemotherapy or HSCT. Using galactomannan (GM) or β-D-glucan (BDG) alone (plus host and CT scan evidence) the apparent incidence of IFD would be 13% and 17% respectively, 8% and 4% lower than the true incidence. The sensitivity, specificity, positive predictive value, and negative predictive value was 63%, 61%, 34%, and 84% for GM and 89%, 21%, 44%, and 75% for BDG respectively. The overall and IFD-attributable mortality rates were 32% (64/203) and 6% (13/203) respectively and this was significantly higher among IFD cases (55% mortality, 33% attributable mortality, P=0.001). The most common causes of death were relapse/progression of underlying disease (38, 59%) and IFD (13, 20%).Our findings demonstrate that a multi-diagnostic approach is necessary in order to improve diagnostic accuracy of IFD. Relying on GM or BDG alone underestimates the true incidence of IFD. Our data suggests that the ‘real world’ incidence of IFD is higher than usually reported especially in tertiary referral transplant units. This data has important implications for clinical practice as well as clinical trials where improved diagnostic tools would improve recruitment.Ceesay: Pfizer, Gilead: Research Funding. Berry:Pfizer, Gilead: Research Funding. Kibbler:Gilead: Consultancy, Research Funding. Wade:Pfizer, Gilead: Research Funding. Smith:Pfizer, Gilead: Research Funding.

The utility of cord blood as an alternative source of stem cells for haematopoietic stem cell transplantation is well established in the paediatric population. Although it's use in adults has surged over the past few years there are still problems associated with a low stem cell dose which may lead to delayed engraftment, incomplete and inefficient immune reconstitution, and poor survival. Various techniques have been used to increase the stem cell dose including ex vivo expansion, the use of double cords, intrabone injections and the use of third party donor cells. Using standard collection techniques only about 25% of cord blood samples collected reach the target cell dose in adults. The placenta is a rich source of CD34+ cells which may not be fully mobilised using current cord blood collection techniques. A modified two-step collection technique in which the initial collection is followed by placental perfusion with 50 ml heparinised 0.9% saline has shown a 15% increase in the number of nucleated cells. We have modified this technique further by perfusing the placenta with Plerixafor, a CXCR4 antagonist that is currently licensed in the UK for mobilisation of autologous stem cells from adults. The study aim was to assess whether plerixafor could enhance recovery of CD34+ cells and change the immune profile of the perfusate.Ten cord blood units were collected as per our standard operating procedures. After cord blood collection, 5 placentas were perfused with 50 ml 0.9% saline plus 5,000 units of preservative free heparin by slow infusion over 50 minutes and perfusate collected after about 5 minutes of starting the infusion over 50 minutes (controls). The other 5 placentas were treated the same way except for the addition of 24 mg Plerixafor to the 50 ml 0.9% saline plus 5,000 units of preservative free heparin and collected as per control arm (plerixafor arm).The samples were analysed by multicolour flow cytometry using a combination of antibody cocktails to identify and enumerate CD34+ stem cells, T cells, T cell precursors, T cell subsets, T regulatory cells, B cells, NK and NK cell precursors, and Dendtritic cells. The CD34+ progenitor cells were further quantified by CFU – a methylcellulose media based assay.Successful recovery of perfusate was achieved from 9 placentas (mean volume 44 ml +/− SEM 4.7) as one of the collected samples clotted. There was no significant difference in the perfusate volumes between the 2 arms (48ml v 38ml, P=0.152). Percentage increase in numbers of cells recovered by perfusing the placenta was calculated from total numbers of CD34+ and CD45+ cells collected from each cord and placenta pair. Plerixafor did not significantly increase the recovery of cells from the placenta. Perfusion of the 5 control placentas increased recovery of CD34+ cell by 5.06% (mean +/− 0.89 SEM) and CD45+ cells by 4.69% (mean +/−1.09 SEM). Perfusion of 4 placentas with saline plus Plerixafor increased recovery of CD34+ cells by 5.79% (mean +/−2.17 SEM) and CD45+ cells by 2.61% (mean +/− 0.79 SEM). Viability of haematopoietic stem cell progenitors was assessed by colony formation assay with no significant difference detected between the control arm (mean 18.4 CFU +/− 8.26 SEM; n=5) and the plerixafor arm (mean 4.8 CFU +/− 2.7 SEM; n=4).There was no significant difference in the number of CD34+ cells recovered from the controls as compared to their corresponding cords (n=5, paired t test, p= 0.054). Similarly, there was no significant difference in the plerixafor arm (n=4, paired t test, p=0.091). There was no significant increase in the number of CD34+ stem cells recovered from the plerixafor group as compared to the control group (unpaired t test, p=0.74). There was no significant increase in the number of CD45+ cells in the plerixafor arm compared to the controls (unpaired t test, p= 0.4). Similarly there was no significant difference in the clonal efficiency of the between the 2 groups (unpaired t test, p=0.2).There were no significant differences in the CD3, CD4, CD8 T cells, Tregs, NK, DCs, progenitor T and NK cells between the 2 groups.In this pilot study plerixafor does not seem to significantly increase the CD34+ stem cells and other cell subsets. It is possible that the duration of infusion was too short to mobilise the stem cells. Further studies are warranted to investigate this further.Off Label Use: Plerixafor for stem cell mobilzation from placenta. Shaw:Therakos, a Johnson and Johnson company: Honoraria, Speakers Bureau.

Abstract 4790

Abstract 2972

Mufti: Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau.

No relevant conflicts of interest to declare.

Abstract 4551

Abstract 1500

Off Label Use: fludarabine, alemtuzumab- used off-label for transplant conditioning. Marsh:Genzyme: Consultancy, Honoraria.

Abstract 2265

The use of reduced intensity conditioning regimens (RIC) has facilitated the allografting of older patients while concurrently reducing the transplant-related mortality associated with allogeneic haematopoietic stem cell transplantation. At present, there is limited data available on the longer-term efficacy and safety of RIC HSCT. We performed a retrospective study on the long term outcomes of patients with haematological malignancies who received RIC HSCT at our center over an 8 year period from Jan 1997 to Dec 2005. A total of 263 adult patients received RIC HSCT, of which 135 patients (51%) survived for 2 or more years and were analysed for long-term outcomes and complications. The median recipient age among our long term survivors at HSCT was 49 years (range: 24–66), with an M: F ratio of 1.6:1 and median follow-up of 51.8 months (range 23.9–100.3). All patients received alemtuzumab (CAMPATH 1H) based RIC regimens with cyclosporin A as post-transplant GvHD prophylaxis. 61 were sibling donor and 74 were volunteer unrelated donor (VUD) transplants. 90 patients (66%) had a myeloid malignancy (mainly AML and MDS), 35 (26%) had lymphoid malignancy and 9 (7%) had myeloma. The stem cell source was PBSC in 98(73%) and bone marrow in 37(27%) cases. Using a 2-year landmark analysis, the actuarial survival at 3 and 5 years was 96% (95% CI: 94.3–97.7) and 91.6% (95% CI: 88.6- 94.6) respectively. There were 8 deaths in the cohort. Four deaths (50%) were due to late relapses, with 4 others due to respiratory failure(1), HBV reactivation with fulminant hepatic failure(1), adenovirus infection(1), chronic GvHD with invasive pulmonary aspergillosis(1). 44 patients had prior acute GvHD (33%). 20(15%) developed chronic GvHD (13 cutaneous involvement, 1 hepatic, 2 hepatic and cutaneous, 5 bronchiolitis obliterans). 18 other patients developed GvHD post donor lymphocytes infusion (DLI). The cumulative incidence of active cGvHD at 3 years and 5 years was 8% and 4% respectively. Significant infectious complications occurred in 38 patients (28%) of which 20 had VZV reactivation most commonly involving the trigeminal nerve. A further 14 had pulmonary problems (9-recurrent chest infections, 2-pneumocystis carinii pneumonia, 2-IPA, 1-TB). Endocrine complications included hyperthyroidism (2 cases) and hypothyroidism (14 cases). In addition, there were 4 cases (8% incidence) of premature menopause and 4 cases (5% incidence) of erectile dysfunction. Secondary malignancy occurred only in 1 patient who developed squamous cell carcinoma of the skin. In addition there were 4 cases of moderate renal impairment, 2 cases of cerebral ischaemic attacks, 8 reports of immune thrombocytopenia/ autoimmune haemolytic anemia. Patients who survive beyond 2 years following alemtuzumab-based RIC HSCT have an excellent overall survival, with a low incidence of chronic GvHD. Nevertheless, late complications of HSCT are still frequent in this cohort. Our data suggests that late effect in patients following RIC HSCT is different from those seen in patients following myeloablative HSCT. Prospective studies are warranted in long-term survivors of RIC HSCT in order to guide appropriate longer-term follow-up.

The use of reduced intensity conditioning regimens (RIC) has facilitated the allografting of older patients while concurrently reducing the transplant-related mortality associated with allogeneic haematopoietic stem cell transplantation. At present, there is limited data available on the longer-term efficacy and safety of RIC HSCT. We performed a retrospective study on the long term outcomes of patients with haematological malignancies who received RIC HSCT at our center over an 8 year period from Jan 1997 to Dec 2005. A total of 263 adult patients received RIC HSCT, of which 135 patients (51%) survived for 2 or more years and were analysed for long-term outcomes and complications. The median recipient age among our long term survivors at HSCT was 49 years (range: 24–66), with an M: F ratio of 1.6:1 and median follow-up of 51.8 months (range 23.9–100.3). All patients received alemtuzumab (CAMPATH 1H) based RIC regimens with cyclosporin A as post-transplant GvHD prophylaxis. 61 were sibling donor and 74 were volunteer unrelated donor (VUD) transplants. 90 patients (66%) had a myeloid malignancy (mainly AML and MDS), 35 (26%) had lymphoid malignancy and 9 (7%) had myeloma. The stem cell source was PBSC in 98(73%) and bone marrow in 37(27%) cases. Using a 2-year landmark analysis, the actuarial survival at 3 and 5 years was 96% (95% CI: 94.3–97.7) and 91.6% (95% CI: 88.6- 94.6) respectively. There were 8 deaths in the cohort. Four deaths (50%) were due to late relapses, with 4 others due to respiratory failure(1), HBV reactivation with fulminant hepatic failure(1), adenovirus infection(1), chronic GvHD with invasive pulmonary aspergillosis(1). 44 patients had prior acute GvHD (33%). 20(15%) developed chronic GvHD (13 cutaneous involvement, 1 hepatic, 2 hepatic and cutaneous, 5 bronchiolitis obliterans). 18 other patients developed GvHD post donor lymphocytes infusion (DLI). The cumulative incidence of active cGvHD at 3 years and 5 years was 8% and 4% respectively. Significant infectious complications occurred in 38 patients (28%) of which 20 had VZV reactivation most commonly involving the trigeminal nerve. A further 14 had pulmonary problems (9-recurrent chest infections, 2-pneumocystis carinii pneumonia, 2-IPA, 1-TB). Endocrine complications included hyperthyroidism (2 cases) and hypothyroidism (14 cases). In addition, there were 4 cases (8% incidence) of premature menopause and 4 cases (5% incidence) of erectile dysfunction. Secondary malignancy occurred only in 1 patient who developed squamous cell carcinoma of the skin. In addition there were 4 cases of moderate renal impairment, 2 cases of cerebral ischaemic attacks, 8 reports of immune thrombocytopenia/ autoimmune haemolytic anemia. Patients who survive beyond 2 years following alemtuzumab-based RIC HSCT have an excellent overall survival, with a low incidence of chronic GvHD. Nevertheless, late complications of HSCT are still frequent in this cohort. Our data suggests that late effect in patients following RIC HSCT is different from those seen in patients following myeloablative HSCT. Prospective studies are warranted in long-term survivors of RIC HSCT in order to guide appropriate longer-term follow-up.

The myelodysplastic syndromes (MDS) encompass a heterogeneous group of clonal stem cell disorders characterised by ineffective erythropoiesis and peripheral blood cytopenias. However, specific subgroups of MDS with both myeloproliferative (MPD) and dysplastic features are recognized, which include chronic myelomonocytic leukaemia (CMML), refractory anaemia with ringed sideroblasts and thrombocytosis (RARS-T) and MDS/MPD unclassified. In addition cases of 5q- syndrome may present with a marked elevation in platelet count and a hypercellular bone marrow. The molecular events which determine the predominant morphological features however remain unclear. The JAK2 somatic point mutation (V617F) results in constitutive activation of the tyrosine kinase and culminates in both proliferation and differentiation signals to the cell and thus may contribute to the proliferative features identified in MDS as reported in the chronic MPDs. We analysed 194 patients from six centres with a diagnosis of MDS or MDS/MPD for the presence of the JAK2 mutation. 11 cases of RA; 70, RARS; 60, 5q- syndrome; 28, refractory anaemia with excess blasts (RAEB); 9, acute myeloid leukaemia with multi-lineage dysplasia (AML-MLD); 4, MDS/MPD-U; 1, CMML; and 1, hypoplastic MDS were analysed. DNA samples were obtained from bone marrow aspirate or peripheral blood. Ethical approval was obtained. Genomic DNA was prepared using standard methods (Qiagen). A modified allele specific PCR (AS PCR) was used to screen DNA samples. JAK2 mutated DNA samples were further subjected to pyrosequencing for confirmatory testing. The median age of the JAK2 mutant versus wild type (WT) cases was 66years (46–80years) v 67years (30–92years). The JAK2 mutation was detected in 13/194 (6.7%) cases overall. 4/70 (5.7%) RARS, 5/60 (8%) 5q- syndrome, 1/28 RAEB (associated with del 5q), 1/4 CMML, 1 MDS/MPD-U and 1/8 AML-MLD demonstrated the mutation. Within the RARS cohort, the prevalence of JAK2 mutation increased to 4/6 (67%) of cases with thrombocytosis (platelets >500x109/l). The presence of the mutation was confirmed in 8/13 cases. In cases analysed, the mutation was detected in CD34+, CD14+, CD15+ and CD61+ cell fractions but not CD3+ or CD19+ cell lineages. On analysis of the haemoglobin (Hb), platelet count and total white cell count (WCC) in the JAK2 mutant versus wild-type groups, there was no difference in median Hb (10.8 v 9.2 g/dl, p=0.275) but a significantly higher median platelet count (526 v 209 x109/l, p<0.001) and WCC (6.43 v 4.84x109/l, P<0.001) was observed in the JAK2 mutant cases. In addition, all JAK2 mutant cases were associated with marked increase in bone marrow cellularity. The JAK2 mutation identifies a subset of MDS patients with proliferative bone marrow morphology and frequent thrombocytosis and leucocytosis. The presence of JAK2 mutation helps to identify a subgroup of MDS patients who may benefit from therapies that specifically target the mutant JAK2.

The myelodysplastic syndromes (MDS) encompass a heterogeneous group of clonal stem cell disorders characterised by ineffective erythropoiesis and peripheral blood cytopenias. However, specific subgroups of MDS with both myeloproliferative (MPD) and dysplastic features are recognized, which include chronic myelomonocytic leukaemia (CMML), refractory anaemia with ringed sideroblasts and thrombocytosis (RARS-T) and MDS/MPD unclassified. In addition cases of 5q- syndrome may present with a marked elevation in platelet count and a hypercellular bone marrow. The molecular events which determine the predominant morphological features however remain unclear. The JAK2 somatic point mutation (V617F) results in constitutive activation of the tyrosine kinase and culminates in both proliferation and differentiation signals to the cell and thus may contribute to the proliferative features identified in MDS as reported in the chronic MPDs. We analysed 194 patients from six centres with a diagnosis of MDS or MDS/MPD for the presence of the JAK2 mutation. 11 cases of RA; 70, RARS; 60, 5q- syndrome; 28, refractory anaemia with excess blasts (RAEB); 9, acute myeloid leukaemia with multi-lineage dysplasia (AML-MLD); 4, MDS/MPD-U; 1, CMML; and 1, hypoplastic MDS were analysed. DNA samples were obtained from bone marrow aspirate or peripheral blood. Ethical approval was obtained. Genomic DNA was prepared using standard methods (Qiagen). A modified allele specific PCR (AS PCR) was used to screen DNA samples. JAK2 mutated DNA samples were further subjected to pyrosequencing for confirmatory testing. The median age of the JAK2 mutant versus wild type (WT) cases was 66years (46–80years) v 67years (30–92years). The JAK2 mutation was detected in 13/194 (6.7%) cases overall. 4/70 (5.7%) RARS, 5/60 (8%) 5q- syndrome, 1/28 RAEB (associated with del 5q), 1/4 CMML, 1 MDS/MPD-U and 1/8 AML-MLD demonstrated the mutation. Within the RARS cohort, the prevalence of JAK2 mutation increased to 4/6 (67%) of cases with thrombocytosis (platelets >500x109/l). The presence of the mutation was confirmed in 8/13 cases. In cases analysed, the mutation was detected in CD34+, CD14+, CD15+ and CD61+ cell fractions but not CD3+ or CD19+ cell lineages. On analysis of the haemoglobin (Hb), platelet count and total white cell count (WCC) in the JAK2 mutant versus wild-type groups, there was no difference in median Hb (10.8 v 9.2 g/dl, p=0.275) but a significantly higher median platelet count (526 v 209 x109/l, p<0.001) and WCC (6.43 v 4.84x109/l, P<0.001) was observed in the JAK2 mutant cases. In addition, all JAK2 mutant cases were associated with marked increase in bone marrow cellularity. The JAK2 mutation identifies a subset of MDS patients with proliferative bone marrow morphology and frequent thrombocytosis and leucocytosis. The presence of JAK2 mutation helps to identify a subgroup of MDS patients who may benefit from therapies that specifically target the mutant JAK2.

ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

Magnetic viscosity and hysteresis versus temperature and field have been studied between room and Curie temperatures on two selected coarse-grained submarine basalts carrying multidomain (MD) low Curie temperature titanomagnetite. These samples exhibit typical characteristics of magnetic viscosity in rocks, including imperfect linearity of the magnetization change with logarithm of time and a large increase in viscosity with temperature, followed by a decrease occurring some 30°–40° below the average Curie point of the rock sample. The field dependence of magnetic viscosity exhibits a maximum located slightly above the sample coercive force. Other experiments, reported elsewhere, show that diffusion plays a negligible role in these rocks. Thus, magnetic viscosity is probably entirely of thermal origin. In spite of the MD characteristics of our samples at room temperature, alternating field (AF) demagnetization of remanence shows, in one of them, the presence of a large component of high coercivity of PSD or SD origin. The remanence of the other sample seems to be wholly of MD origin. The thermal dependences of magnetic viscosity and irreversible susceptibility are only approximately similar, and the field dependences are quite different. This indicates that the proportionality of irreversible susceptibility with magnetic viscosity predicted by the Néel and the Street and Wooley theories of thermal fluctuations in MD grains does not hold for natural titanomagnetite like ours, even though it is verified for synthetic ferromagnetic substances. However, our experimental data are compatible with the fundamentals of the theory of thermal fluctuations if we assume that the pinning of Bloch walls in natural titanomagnetite is due to several kinds of energy barriers with a broad distribution of both activation volumes va and critical fields hc. Thus, the remanent magnetization acquired in a low field (1 Oe) applied for 1 d at room temperature is found to be pinned by two categories of defects with (1) va= 7×10-15cm3 and hc= 3Oe and (2) va= 1×10-15cm3 and hc= 15Oe. Using a (va, hc) diagram to represent each barrier, we show that the ‘blocking’(or ‘unblocking’) curves are quite dissimilar for field and time effects, just as for SD particles. This indicates that the proportionality between magnetic viscosity and irreversible susceptibility requires particular distributions of energy barriers in the (va, hc) plane and cannot be considered as a general characteristic of thermally activated magnetic viscosity.