bluuebunny/crossref_metadata_2025_split huggingface dataset:
Leucodepletion for hyperleucocytosis – first report on a novel technology featuring electronic interphase management
Authors: Schulz M. , Bug G. , Bialleck H. , Serve H. , Seifried E. , Bönig H. 
Published: February 2013

Background and ObjectivesTherapeutic leucodepletion plays an established role in the initial treatment of patients with acute myeloid leukaemia (AML) and possibly other leukaemias presenting with leucostasis. Recently, a new leucodepletion technology, Spectra Optia IDL, has become available that differs from its predecessor, COBE Spectra MNC, by a variety of electronic supports, including by electronic adjustment of buffy coat positioning at the collection port. Given the paucity of patients in need of leucodepletions and marked differences in clinical presentation as well as blast properties (e.g. size, density), formal clinical trials comparing leucodepletion technologies have never been executed.Materials and MethodsHere, we present aggregate data from eight leucodepletions performed in AML patients with clinical signs of leucostasis between 11/2011 and 07/2012 with the new device and compare the apheresis outcomes with those from fifteen leucodepletions performed with the old technology between 06/2010 and 10/2011.ResultsPatients did not differ with respect to epidemiological data. Pre‐apheresis leucocyte count (WBC) was significantly higher in Spectra Optia IDL patients. Tolerability was excellent with both devices. Basic apheresis denominators such as duration, processed volume, inlet pump rate, ACD‐A consumption and product volume were very similar. A negative correlation between pre‐apheresis WBC and collection efficiency was noted. Mean collection efficiency for leucocytes with Spectra Optia IDL (47·3%) was similar to that with COBE Spectra MNC (50·5%). Platelet attrition was similar with both devices, approximately 30%.ConclusionThe novel, electronically guided leukapheresis system is suitable for leucodepletion.
Effect of TNF‐alpha blockade on coagulopathy, survival and endothelial activation in xenoperfused porcine kidneys
Authors: Ramackers Wolf , Friedrich Lars , Tiede Andreas , Bergmann Sabine , Bockmeyer Clemens , Schuettler Wolfgang , Becker Jan , Winkler Michael 
Published: February 2013

Background:  Following pig to primate kidney transplantation, endothelial cell activation and xenogenic activation of the coagulation (XAC) system of the recipient eventually leading to organ dysfunction and disseminated intravascular coagulation (DIC) can be observed (1). In this study we examined the effect of a TNF‐Receptor‐Fusionprotein (TNF‐RFP) on XAC and endothelial cell activation utilizing an appropriate ex‐vivo perfusion system.Methods:  Using an ex‐vivo perfusion circuit based on C1‐Inhibitor (C1‐Inh) and low dose heparin administration, we have analysed XAC following contact of human blood with porcine endothelium. Porcine kidneys were recovered following in situ cold perfusion with HTK organ preservation solution and were immediately connected to a perfusion circuit utilizing freshly drawn pooled porcine or human AB blood. The experiments were performed in three individual groups: autologous perfusion (n = 5), xenogenic perfusion without any further pharmacological intervention (n = 10) or with addition of TNF‐RFP (n = 5). After perfusion tissue samples were obtained for QPCR and immunohistological analyses. Endothelial cell activation was assessed by measuring the expression levels of E‐Selectin, ICAM‐1 and VCAM‐1 in QPCR.Results:  Kidney survival during organ perfusion with human blood, CI‐Inh, heparin but without any further pharmacological intervention was 126 ± 78 min. XAC was observed with significantly elevated concentrations of d‐Dimer, thrombin antithrombin complex (TAT), resulting in formation of multiple microthrombi. Endothelial cell activation was pronounced, as shown by increased expression of E‐Selectin and VCAM‐1. In contrast, pharmacological intervention with TNF‐RFP reduced the consumption of antithrombin and fibrinogen and prolonged organ survival to 240 min (±0). Additionally, formation of microtrombi was reduced compared to the xenogenic control without pharmacological intervention. Remarkably, also endothelial cell activation was significantly reduced in the TNF‐RFP group.Conclusions:  We conclude that TNF‐RFP can interfere with XAC and is able to suppress endothelial cell activation in this ex vivo perfusion model, thus representing a potential candidate as an adjuvant immunosuppressive drug.Reference:  1. BÜhler L, Basker M, Alwayn IPet al. Coagulation and thrombotic disorders associated with pig organ and hematopoietic cell transplantation in nonhuman primates. Transplantation 2000; 9: 1323–1331.
Evidence of safety and efficacy of two forms of neonatal cell transplants
Authors: Elliott Robert B. , Nathu Divya 
Published: February 2013

On behalf of DOL and LCT  A clinical trial in 14 adult type 1 diabetics intra‐peritoneal alginate microencapsulated neonatal porcine islets transplants commenced in NZ over 2 years ago and the last patient has just completed the required 12 months of follow up.To meet regulatory requirements for xenotransplantation, an elaborate program on xenovirology has been developed on one unique herd of pigs that was free from all conventional pathogens and was qualified as “null” pigs, i.e. they do not have a transmittable pig endogenous retrovirus (PERV). These pigs are the animal founders for the DPF donor herd. A comprehensive program for patients’ microbiology follow‐up was also developed. This program includes assays developed for monitoring potential infection with PERV and other potentially xenotic pathogens.All of the patients selected had unaware hypoglycaemia. The dose of islets used varied from 5000/kg (n = 4) 10 000/kg (n = 4) 15 000/kg (n = 4), and 20 000/kg (n = 2). The patients experience little if any side effects and there was no evidence of xenosis. Unaware hypoglycaemia was lessened in all groups but more so in the 10 000/kg group. Insulin dose was lowered and HbA1c improved in some but not all subjects. The patients with maximum insulin dose drop received 5000/kg dose. There was no clear cut relationship of return of hypoglycaemia awareness and glucagon or adrenergic responses to induced hypoglycaemia. There was direct laboratory evidence of islet beta cell function. Clinical follow up at 2 years of some patients indicates persistence of the benefits. A further clinical trial involving a different dosing schedule and some technical improvements is in progress.Choroid plexus (CP) ependymal epithelium secretes a wide variety of neurotrophic and neural support molecules. Micro‐encapsulated CP delivered intra‐cerebrally into rat models of stroke, Huntington's disease and Parkinson's disease show remarkable functional and histological recovery. In the quinolinic acid primate model of Huntington's and the MTPP model of Parkinson's functional recovery and relevant histological evidence of neuro‐regeneration were seen. The treatment was safe. A clinical trial of this form of treatment in humans with Parkinson's is being undertaken.Conclusions:  Xenotransplantation of microencapsulated islets and other cell types into human and non‐human primates from a biocertified herd of pigs prepared under GMP conditions can be conducted with safety and efficacy. Further improvements in clinical outcomes are expected with technical advances.
Structural and Functional Analysis of the Pore-Forming Toxin NetB from Clostridium perfringens
Authors: Yan Xu-Xia , Porter Corrine J. , Hardy Simon P. , Steer David , Smith A. Ian , Quinsey Noelene S. , Hughes Victoria , Cheung Jackie K. , Keyburn Anthony L. , Kaldhusdal Magne , Moore Robert J. , Bannam Trudi L. , Whisstock James C. , Rood Julian I. 
Published: February 2013

ABSTRACT

Clostridium perfringens
is an anaerobic bacterium that causes numerous important human and animal diseases, primarily as a result of its ability to produce many different protein toxins. In chickens,
C. perfringens
causes necrotic enteritis, a disease of economic importance to the worldwide poultry industry. The secreted pore-forming toxin NetB is a key virulence factor in the pathogenesis of avian necrotic enteritis and is similar to alpha-hemolysin, a β-barrel pore-forming toxin from
Staphylococcus aureus
. To address the molecular mechanisms underlying NetB-mediated tissue damage, we determined the crystal structure of the monomeric form of NetB to 1.8 Å. Structural comparisons with other members of the alpha-hemolysin family revealed significant differences in the conformation of the membrane binding domain. These data suggested that NetB may recognize different membrane receptors or use a different mechanism for membrane-protein interactions. Consistent with this idea, electrophysiological experiments with planar lipid bilayers revealed that NetB formed pores with much larger single-channel conductance than alpha-hemolysin. Channel conductance varied with phospholipid net charge. Furthermore, NetB differed in its ion selectivity, preferring cations over anions. Using hemolysis as a screen, we carried out a random-mutagenesis study that identified several residues that are critical for NetB-induced cell lysis. Mapping of these residues onto the crystal structure revealed that they were clustered in regions predicted to be required for oligomerization or membrane binding. Together these data provide an insight into the mechanism of NetB-mediated pore formation and will contribute to our understanding of the mode of action of this important toxin.

IMPORTANCE
Necrotic enteritis is an economically important disease of the worldwide poultry industry and is mediated by
Clostridium perfringens
strains that produce NetB, a β-pore-forming toxin. We carried out structural and functional studies of NetB to provide a mechanistic insight into its mode of action and to assist in the development of a necrotic enteritis vaccine. We determined the structure of the monomeric form of NetB to 1.8 Å, used both site-directed and random mutagenesis to identify key residues that are required for its biological activity, and analyzed pore formation by NetB and its substitution-containing derivatives in planar lipid bilayers.
RidA Proteins Prevent Metabolic Damage Inflicted by PLP-Dependent Dehydratases in All Domains of Life
Authors: Lambrecht Jennifer A. , Schmitz George E. , Downs Diana M. 
Published: February 2013

ABSTRACT

Pyridoxal 5′-phosphate (PLP) is a coenzyme synthesized by all forms of life. Relevant to the work reported here is the mechanism of the PLP-dependent threonine/serine dehydratases, which generate reactive enamine/imine intermediates that are converted to keto acids by members of the RidA family of enzymes. The RidA protein of
Salmonella enterica
serovar Typhimurium LT2 is the founding member of this broadly conserved family of proteins (formerly known as YjgF/YER057c/UK114). RidA proteins were recently shown to be enamine deaminases. Here we demonstrate the damaging potential of enamines in the absence of RidA proteins. Notably,
S. enterica
strains lacking RidA have decreased activity of the PLP-dependent transaminase B enzyme IlvE, an enzyme involved in branched-chain amino acid biosynthesis. We reconstituted the threonine/serine dehydratase (IlvA)-dependent inhibition of IlvE
in vitro
, show that the
in vitro
system reflects the mechanism of RidA function
in vivo
, and show that IlvE inhibition is prevented by RidA proteins from all domains of life. We conclude that 2-aminoacrylate (2AA) inhibition represents a new type of metabolic damage, and this finding provides an important physiological context for the role of the ubiquitous RidA family of enamine deaminases in preventing damage by 2AA.

IMPORTANCE
External stresses that disrupt metabolic components can perturb cellular functions and affect growth. A similar consequence is expected if endogenously generated metabolites are reactive and persist in the cellular environment. Here we show that the metabolic intermediate 2-aminoacrylate (2AA) causes significant cellular damage if allowed to accumulate aberrantly. Furthermore, we show that the widely conserved protein RidA prevents this accumulation by facilitating conversion of 2AA to a stable metabolite. This work demonstrates that the reactive metabolite 2AA, previously considered innocuous in the cell due to a short half-life in aqueous solution, can survive in the cellular environment long enough to cause damage. This work provides insights into the roles and persistence of reactive metabolites
in vivo
and shows that the RidA family of proteins is able to prevent damage caused by a reactive intermediate that is created as a consequence of PLP-dependent chemistry.
Carbon Catabolite Control in Candida albicans: New Wrinkles in Metabolism
Authors: Lorenz Michael C. 
Published: February 2013

ABSTRACT

Most microorganisms maintain strict control of nutrient assimilation pathways to ensure that they preferentially use compounds that generate the most energy or are most efficiently catabolized. In doing so, they avoid potentially inefficient conflicts between parallel catabolic and metabolic pathways. The regulation of carbon source utilization in a wide array of bacterial and fungal species involves both transcriptional and posttranscriptional mechanisms, and while the details can vary significantly, carbon catabolite control is widely conserved. In many fungi, the posttranslational aspect (carbon catabolite inactivation [CCI]) involves the ubiquitin-mediated degradation of catabolic enzymes for poor carbon sources when a preferred one (glucose) becomes available. A recent article presents evidence for a surprising exception to CCI in the fungal pathogen
Candida albicans
, an organism that makes use of gluconeogenic carbon sources during infection (D. Sandai, Z. Yin, L. Selway, D. Stead, J. Walker, M. D. Leach, I. Bohovych, I. V. Ene, S. Kastora, S. Budge, C. A. Munro, F. C. Odds, N. A. Gow, and A. J. Brown,
mBio
3[6]:e00495-12).
In vitro
, addition of glucose to cells grown in a poor carbon source rapidly represses transcripts encoding gluconeogenic and glyoxylate cycle enzymes, such as phosphoenolpyruvate carboxykinase (Pck1p) and isocitrate lyase (Icl1p), in both
C. albicans
and
Saccharomyces cerevisiae
. Yet, uniquely, the
C. albicans
proteins persist, permitting parallel assimilation of multiple carbon sources, likely because they lack consensus ubiquitination sites found in the yeast homologs. Indeed, the yeast proteins are rapidly degraded when expressed in
C. albicans
, indicating a conservation of the machinery needed for CCI. How this surprising metabolic twist contributes to fungal commensalism or pathogenesis remains an open question.
Next-Generation Sequencing of Small RNAs from HIV-Infected Cells Identifies Phased microRNA Expression Patterns and Candidate Novel microRNAs Differentially Expressed upon Infection
Authors: Chang Stewart T. , Thomas Matthew J. , Sova Pavel , Green Richard R. , Palermo Robert E. , Katze Michael G. 
Published: February 2013

ABSTRACT

HIV infection of CD4
+
T cells induces a range of host transcriptional changes in mRNAs as well as microRNAs that may coordinate changes in mRNAs. To survey these dynamic changes, we applied next-generation sequencing, analyzing the small RNA fraction of HIV-infected cells at 5, 12, and 24 h postinfection (RNA-Seq). These time points afforded a view of the transcriptomic changes occurring both before and during viral replication. In the resulting small RNA-Seq data set, we detected a phased pattern of microRNA expression. Largely distinct sets of microRNAs were found to be suppressed at 5 and 12 h postinfection, and both sets of changes rebounded later in infection. A larger set of microRNA changes was observed at 24 h postinfection. When integrated with mRNA expression data, the small RNA-Seq data indicated a role for microRNAs in transcriptional regulation, T cell activation, and cell cycle during HIV infection. As a unique benefit of next-generation sequencing, we also detected candidate novel host microRNAs differentially expressed during infection, including one whose downregulation at 24 h postinfection may allow full replication of HIV to proceed. Collectively, our data provide a uniquely comprehensive view of the changes in host microRNAs induced by HIV during cellular infection.

IMPORTANCE
New sequencing technologies allow unprecedented views into changes occurring in virus-infected cells, including comprehensive and largely unbiased measurements of different types of RNA. In this study, we used next-generation sequencing to profile dynamic changes in cellular microRNAs occurring in HIV-infected cells. The sensitivity afforded by sequencing allowed us to detect changes in microRNA expression early in infection, before the onset of viral replication. A phased pattern of expression was evident among these microRNAs, and many that were initially suppressed were later overexpressed at the height of infection, providing unique signatures of infection. By integrating additional mRNA data with the microRNA data, we identified a role for microRNAs in transcriptional regulation during infection and specifically a network of microRNAs involved in the expression of a known HIV cofactor. Finally, as a distinct benefit of sequencing, we identified candidate nonannotated microRNAs, including one whose downregulation may allow HIV-1 replication to proceed fully.
Air pollution exposure in early pregnancy and adverse pregnancy outcomes: a register-based cohort study
Authors: Olsson David , Mogren Ingrid , Forsberg Bertil 
Published: February 2013

ObjectivesOur aim was to study the possible associations between exposure to elevated levels of air pollution, ozone (O3) and vehicle exhaust (NOx), during early gestation, and adverse pregnancy outcomes such as pre-eclampsia, preterm birth and small for gestational age.DesignProspective register-based cohort study.SettingThe Swedish Medical Birth Register includes data on all deliveries during 1998 to 2006 in Greater Stockholm, Sweden. The national Patient Register and the Prescribed Drug Register were used to collect information on maternal asthma.ParticipantsAll singleton pregnancies, conceived at the earliest in August 1997 and at the latest in February 2006, were included, n=120 755.Outcome measuresWe studied preterm birth, small for gestational age and pre-eclampsia.Results4.4% of pregnancies resulted in a preterm birth. The prevalence of pre-eclampsia was 2.7%. We observed an association between first trimester O3and preterm birth (OR 1.04, 95% CI 1.01 to 1.08) as well as an association with pre-eclampsia (OR 1.04, 95% CI 1.01 to 1.08), per 10 µg/m3increase in O3. We observed no association between first trimester NOxand adverse pregnancy outcomes. No associations were observed between any of the air pollutants and small for gestational age.ConclusionsIncreased levels of O3during the first trimester increased the risk of pre-eclampsia and preterm birth. Air pollutants did not exhibit any effects on fetal growth restriction. We estimated 1 in every 20 cases of pre-eclampsia to be associated with O3exposure.
A cross-sectional study of workplace social capital and blood pressure: a multilevel analysis at Japanese manufacturing companies
Authors: Fujino Yoshihisa , Kubo Tatsuhiko , Kunimoto Masamizu , Tabata Hidetoshi , Tsuchiya Takuto , Kadowaki Koji , Nakamura Takehiro , Oyama Ichiro 
Published: February 2013

ObjectivesWe examined the contextual effect of workplace social capital on systolic blood pressure (SBP).DesignCross-sectional.SettingA conglomerate from 58 workplaces in Japan.ParticipantsOf the 5844 workers at a Japanese conglomerate from 58 workplaces, 5368 were recruited. Individuals who received drugs for hypertension (n=531) and who lacked information on any variable (n=167) were excluded from the analyses, leaving 4735 individuals (3281 men and 1454 women) for inclusion.Primary and secondary outcome measuresSystolic blood pressure.ResultsThe contextual effect of workplace social capital on SBP was examined using a multilevel regression analysis with a random intercept. Coworker support had a contextual effect at the workplace level (coefficient=−1.97, p=0.043), while a lack of trust for coworkers (coefficient=0.27, p=0.039) and lack of helpfulness from coworkers were associated with SBP (coefficient=0.28, p=0.002).ConclusionsThe present study suggested that social capital at the workplace level has beneficial effects on SBP.
Treatment quality indicators predict short-term outcomes in patients with diabetes: a prospective cohort study using the GIANTT database
Authors: Sidorenkov Grigory , Voorham Jaco , Zeeuw Dick de , Haaijer-Ruskamp Flora M , Denig Petra 
Published: February 2013

ObjectiveTo assess whether quality indicators for treatment of cardiovascular and renal risk factors are associated with short-term outcomes in patients with diabetes.DesignA prospective cohort study using linear regression adjusting for confounders.SettingThe GIANTT database (Groningen Initiative to Analyse Type 2 Diabetes Treatment) containing data from primary care medical records from The Netherlands.Participants15 453 patients with type 2 diabetes mellitus diagnosed before 1 January 2008. Mean age 66.5 years, 47.5% men.ExposureQuality indicators assessing current treatment (CT) status or treatment intensification (TI) for patients with diabetes with elevated cardiovascular or renal risk factors.Main outcome measuresLow-density lipoprotein cholesterol (LDL-C), systolic blood pressure (SBP), and albumin:creatinine ratio (ACR) before and after assessment of treatment quality.ResultsUse of lipid-lowering drugs was associated with better LDL-C levels (−0.41 mmol/litre; 95% CI −0.48 to −0.34). Use of blood pressure-lowering drugs and use of renin–angiotensin system inhibitors in patients with elevated risk factor levels was not associated with better SBP and ACR outcomes, respectively. TI was also associated with better LDL-C (−0.82 mmol/litre; CI −0.93 to −0.71) in patients with elevated LDL-C levels, and with better SBP (−1.26 mm Hg; CI −2.28 to −0.24) in patients with two elevated SBP levels. Intensification of albuminuria-lowering treatment showed a tendency towards better ACR (−2.47 mmol/mg; CI −5.32 to 0.39) in patients with elevated ACR levels.ConclusionsQuality indicators of TI were predictive of better short-term cardiovascular and renal outcomes, whereas indicators assessing CT status showed association only with better LDL-C outcome.