bluuebunny/crossref_metadata_2025_split huggingface dataset:
Two steps to functional mesenchymal stromal cells for clinical application
Authors: Bartmann Christina , Rohde Eva , Schallmoser Katharina , Pürstner Peter , Lanzer Gerhard , Linkesch Werner , Strunk Dirk 
Published: May 2007

BACKGROUND: Ex vivo expansion of multipotent mesenchymal stromal cells (MSCs) is a prerequisite for evaluating their therapeutic potential in ongoing clinical trials. Even large volumes of starting material and extended culture periods, however, do not necessarily produce 2 × 106 MSCs per kg per adult patient. A new two‐step procedure has been devised to propagate more than 1 × 108 MSCs from small marrow volumes within fewer than 4 weeks.STUDY DESIGN AND METHODS: The influence of log fold decreased MSC seeding (2500, 250, 25, 2.5/cm2) on clinical‐scale expansion, MSC phenotype, and immunomodulatory function combined with multiplex cytokine display was analyzed. Maintenance of MSC characteristics was tested in fibroblast colony‐forming unit and differentiation assays.RESULTS: Reduced seeding density boosted MSC propagation. Low‐density expanded MSCs were CD29+, CD73+, CD90+, CD105+, CD14–, CD34–, CD45–, HLA‐DR–; retained their differentiation potential; and inhibited lymphocyte proliferation. This was accompanied by deregulated cytokine production. Seeding 0.7 × 106 to 1 × 106 MSCs derived from a 10‐ to 13‐day primary culture at a low density of 28 to 40 per cm2 permitted propagation of 1.5 × 108 to 3.7 × 108 functional MSCs within a 13‐ to 15‐day secondary expansion step.CONCLUSION: Primary seeding of only 10‐mL marrow aspirates on approximately 0.2‐m2 culture area (Step 1) followed by expansion on 2.5 m2 (Step 2) is sufficient to consistently generate at least 1.5 × 108 MSCs in fetal bovine serum–supplemented medium within less than 4 weeks. The efficiency of this two‐step procedure for clinical‐scale MSC propagation may facilitate rational clinical testing of MSC‐based therapies.
Human platelet lysate can replace fetal bovine serum for clinical‐scale expansion of functional mesenchymal stromal cells
Authors: Schallmoser Katharina , Bartmann Christina , Rohde Eva , Reinisch Andreas , Kashofer Karl , Stadelmeyer Elke , Drexler Camilla , Lanzer Gerhard , Linkesch Werner , Strunk Dirk 
Published: May 2007

BACKGROUND: Human multipotent mesenchymal stromal cells (MSCs) are promising candidates for a growing spectrum of regenerative and immunomodulatory cellular therapies. Translation of auspicious experimental results into clinical applications has been limited by the dependence of MSC propagation from fetal bovine serum (FBS).STUDY DESIGN AND METHODS: The capacity of human platelet lysate (HPL) to replace FBS for clinical‐scale MSC propagation was analyzed.RESULTS: HPL could be efficiently produced from buffy coats. Multiplex analyses allowed a distinct HPL growth factor profile to be delineated. With a previously established two‐step clinical‐scale procedure, HPL was reproducibly more efficient than FBS in supporting MSC outgrowth. With only 3 × 105 primary culture‐derived MSCs, a mean of 4.36 × 108 HPL‐MSCs (range, 3.01 × 108‐5.40 × 108) was obtained within a single secondary 11‐ to 13‐day culture step. Although morphologically distinct, HPL‐MSCs and FBS‐MSCs did not differ significantly in terms of immunophenotype, differentiation potential in vitro, and lack of tumorigenicity in nude mice in vivo.CONCLUSIONS: Replacing FBS with HPL prevents bovine prion, viral, and zoonose contamination of the stem cell product. This new efficient FBS‐free two‐step procedure for clinical‐scale MSC propagation may represent a major step toward challenging new stem cell therapies.
Cryopreservation of cellular products in a closed‐bag system with an incorporated dimethyl sulfoxide–resistant sterile filter outside of cleanroom facilities
Authors: Humpe Andreas , Jansen Pal , Tischer Bernd Karsten , Schubert Sabine , Beck Christian , Adamzik Ilse‐Dorothea , Maas Jens‐Holger , Strate Alexander , Gramatzki Martin , Riggert Joachim 
Published: May 2007

BACKGROUND: Manipulations, for example, cryopreservation, of cellular therapeutics carried out in an open system must be performed in a class A environment with surrounding class B environment. To avoid cleanroom facilities, a new closed‐bag system with an incorporated dimethyl sulfoxide–resistant sterile filter for cryopreservation of cellular products was evaluated at two different centers.STUDY DESIGN AND METHODS: A total of 44 different products (22 buffy coats [BCs] and 22 leukapheresis [LK] products) were split and cryopreserved in parallel in cleanroom facilities (Method I) and with the closed system on the bench of a “normal” laboratory (Method II). Viability analyzed by 7‐aminoactinomycin D staining and flow cytometric analysis and sterility of the products were analyzed.RESULTS: Independent of the cellular source (BC or LK), the median viability of CD45+ cells decreased significantly (p  < 0.01) during cryopreservation: namely, in BCs, −15.8 percent with both methods, and in LK products, −5.4 percent with Method I and −4.8 percent with Method II, respectively. CD3+ as well as CD14+ cells exhibited a similar pattern and were also found significantly (p  < 0.01) diminished after thawing independent of the handling system. For CD19+ cells, the small decrease of viability was only for the BC group significant (p = 0.027) when the cells had been processed with Method I. No bacterial contamination was detected neither in fresh products nor in products after cryopreservation.CONCLUSION: The closed system for cryopreservation of cellular products appears to be equivalent to cleanroom‐based methods regarding cellular integrity and sterility when appropriate quality of sterile filters is assured.
Protein A immunoadsorption therapy for refractory, mitomycin C–associated thrombotic microangiopathy
Authors: Kasper Stefan , Neurath Markus F. , Huber Christoph , Theobald Matthias , Scharrer Inge 
Published: May 2007

BACKGROUND: Mitomycin C–associated thrombotic microangiopathy (TMA) has a poor prognosis with limited therapeutic options. Most patients die within 4 months of diagnosis due to pulmonary or renal failure. Here, a patient resistant to total plasma exchange (TPE) and immunosuppressive therapy with glucocorticoids, rituximab, vincristine, and splenectomy who was successfully treated with protein A immunoadsorption is described.CASE REPORT: A 29‐year‐old woman developed a TMA after chemotherapy with mitomycin C. She presented with thrombocytopenia, pulmonary edema, hemolytic anemia with presence of schistocytes, and renal failure. Immediate TPE (>120 times) and immunosuppressive therapy with glucocorticoids, however, did not improve her clinical situation. Furthermore, she was refractory to subsequent immunosuppressive therapy with rituximab and vincristine and laparoscopic splenectomy. Finally, after 12 cycles of extracorporeal protein A immunoadsorption with a commercially available immunoadsorption system (Immunosorba, Fresenius AG), platelet counts increased with disappearance of hemolytic syndromes.CONCLUSION: Extracorporeal protein A immunoadsorption with the Immunosorba system emerges as a potentially effective and safe treatment for refractory mitomycin C–associated TMA with only moderate side effects. This therapeutic option may be considered at an early state of the disease to prevent extensive immunosuppression.
Occurrence and severity of adverse events after autologous hematopoietic progenitor cell infusion are related to the amount of granulocytes in the apheresis product
Authors: Calmels Boris , Lemarié Claude , Esterni Benjamin , Malugani Caroline , Charbonnier Aude , Coso Diane , Schiano de Colella Jean‐Marc , Deconinck Eric , Caillot Denis , Viret Frédéric , Ladaique Patrick , Lapierre Valérie , Chabannon Christian 
Published: May 2007

BACKGROUND: Adverse events (AEs) after hematopoietic progenitor cell (HPC) infusion are rare but might be life‐threatening. These reactions have traditionally been associated with the amount of infused cryoprotectant, but persistence of such events after dimethyl sulfoxide (DMSO) depletion has questioned this assumption.STUDY DESIGN AND METHODS: The incidence of AEs on a cohort of 460 patients (490 HPC infusions) undergoing autologous DMSO‐reduced HPC transplantation was prospectively evaluated. HPCs were collected from adult patients with various hematologic or solid malignancies. After quality control (QC) on fresh apheresis products and subsequent cryopreservation, HPC grafts were thawed and washed at the cell therapy facility. QC was performed on each graft after washing, and clinical data were collected for each infusion.RESULTS: AEs were reported in 66 cases (13.5%) and were graded according to the NCI‐CTC scale from 1 to 4. Although none of the factors associated with patient characteristics or infusion procedure were different between the two groups (no AE vs. occurrence of AE), it was found that the absolute number of granulocytes measured before freezing was considerably higher in the AE group. Furthermore, within this group, there was a strong correlation between the amount of granulocytes and the grading of the reaction.CONCLUSION: This survey demonstrates that AEs occurring in the setting of DMSO‐reduced HPC grafts are directly related to the amount of granulocytes and thus emphasizes the need for high‐quality apheresis products so as to improve the safety of HPC infusion.
Nonleukoreduced red blood cell transfusion induces a sustained inhibition of neutrophil chemotaxis by stimulating in vivo production of transforming growth factor‐β1 by neutrophils: role of the immunoglobulinlike transcript 1, sFasL, and sHLA‐I
Authors: Ottonello Luciano , Ghio Massimo , Contini Paola , Bertolotto Maria , Bianchi Giordano , Montecucco Fabrizio , Colonna Marco , Mazzei Clemente , Dallegri Franco , Indiveri Franco 
Published: May 2007

BACKGROUND: Red blood cell (RBC) transfusion has been linked to increased susceptibility to infections in critically ill patients and to augmented incidence of postoperative infections. The mechanisms by which transfusions can induce immunosuppression are only partially defined. Recently, it has been demonstrated that RBC supernatants inhibit neutrophil migration. Such inhibitory activity is due to transforming growth factor (TGF)‐β1 contained in the supernatants that desensitize neutrophils to subsequent chemotaxic stimulation.STUDY DESIGN AND METHODS: In ancillary experiments, it was observed that plasma from transfused patients maintained its capacity of inhibiting neutrophil chemotaxis several days after RBC transfusion. Thus, this study was planned to investigate the mechanism(s) responsible for the prolonged inhibition of neutrophil chemotaxis observed after RBC transfusion.RESULTS: Plasma samples obtained from subjects who underwent RBC transfusion display a capability of inhibiting neutrophil chemotaxis, which is detectable up to 15 days after the transfusion. The inhibition is related to the capacity of FasL and HLA‐I molecules contained in RBC supernatants to induce in vivo TGF‐β1 synthesis by neutrophils. The induction of TGF‐β1 secretion in neutrophils by HLA‐I molecules depends on immunoglobulinlike transcript 1/CD85 triggering.CONCLUSION: The property of RBC transfusion of inducing a sustained inhibition of neutrophil chemotaxis seems to be a potential mechanism that concurs to the susceptibility to infections in patients who receive transfusions. Furthermore, our findings, showing neutrophil production of TGF‐β1 in response to FasL and HLA‐I molecules, confirm that neutrophils are endowed not only with effector functions but also with immunomodulatory properties possibly involved in the regulation of inflammatory processes.
Genotype probabilities of pairs of individuals for X‐chromosome markers
Authors: Toni Chiara , Domenici Ranieri , Presciuttini Silvano 
Published: May 2007

BACKGROUND: The usual set of autosomal markers (A‐STRs) available in commercial kits is often insufficient to discriminate between close relationships when only two subjects are available for analysis. X‐chromosome markers (X‐STRs) provide higher statistical power in special cases.STUDY DESIGN AND METHODS: Formulas are derived for the probabilities of all possible genotype pairs for X‐STRs of any sex combination for seven common relationships. The power of exclusion (PE) of X‐STRs in parentage analysis is compared with that of A‐STRs of equivalent distribution of allele frequency.RESULTS: Seventy‐three equations were obtained, from which the likelihood ratio of any two alternative hypotheses about the relationship between two individuals can be obtained by division and simplification. For father‐daughter and mother‐son duos, the PE of X‐STRs is almost twice the corresponding value of A‐STRs for moderately low values of heterozygosity (0.6‐0.75); for alleged pairs of sisters and pairs of half‐sisters the PE is equivalent to that of A‐STRs in parent‐child duos. Considering four real unlinked X‐STRs, the cumulative PE for father‐daughter and mother‐son duos was 99 percent, compared with 94 percent if they were autosomal.CONCLUSIONS: X‐STRs can substantially increase the discrimination capacity of standard A‐STRs in parentage analyses involving pairs of individuals. Up to four unlinked X‐STRs may be treated as independent loci. When linked loci are included, computer programs that calculate pedigree likelihoods can be used.
Identification and quantification of fetal red blood cells in maternal blood by a dual‐color flow cytometric method: evaluation of the Fetal Cell Count kit
Authors: Porra Valérie , Bernaud Janine , Gueret Pierre , Bricca Pascaline , Rigal Dominique , Follea Gilles , Blanchard Dominique 
Published: May 2007

BACKGROUND: As an alternative to the cumbersome Kleihauer‐Betke test (KBT), flow cytometry represents a powerful method for the identification and quantification of fetal red blood cells (RBCs) in maternal circulation.STUDY DESIGN AND METHODS: The aim of this study was to evaluate the Fetal Cell Count kit (IQ Products), an innovative flow cytometric method, based on the combination of antibodies directed, respectively, against fetal hemoglobin (HbF) and carbonic anhydrase (CA), a marker expressed after birth, to discriminate fetal RBCs from adult F cells containing HbF. The investigation was performed by two French laboratories that compared the data obtained by flow cytometry and KBT in 455 pregnant or just‐delivered women as well as in 124 artificial mixtures containing from 0.01 to 5.00 percent cord cells.RESULTS: The FL1/FL2 histogram allowed distinction between fetal RBCs (HbF+, CA–), F cells (HbF+, CA+), and adult RBCs (HbF–, CA+). The limits of detection and quantification were determined at 0.03 and 0.10 percent or 0.02 and 0.05 percent when analyzing 100,000 or 200,000 events, respectively. Linearity was demonstrated between 0.01 and 5.00 percent fetal cells in the mixtures (r = 0.95, p < 0.01). A good correlation between fluorescence‐activated cell sorting (FACS) and KBT results was obtained with artificial mixtures (r = 0.94, p < 0.01). From the 405 Kleihauer‐negative samples, none were identified as positive by FACS. Among the 50 Kleihauer‐positive samples, 6 were shown not to contain fetal cells but F cells by FACS.CONCLUSION: With this new dual‐color flow cytometric method, accurate evaluation of fetomaternal hemorrhage was achieved even in the face of HbF of maternal origin.
Do patients with autoantibodies or clinically insignificant alloantibodies require an indirect antiglobulin test crossmatch?
Authors: Lee Edmond , Redman Martin , Burgess Gordon , Win Nay 
Published: May 2007

BACKGROUND: Compatibility testing is the standard protocol that identifies suitable blood for patients requiring transfusion. If the antibody screen is negative or no clinically significant antibodies are detected, BCSH guidelines and AABB standards allow an immediate‐spin crossmatch (IS XM) or even electronic issue. The testing requirement is less clear where autoantibodies or non–clinically significant alloantibodies compromise the indirect antiglobulin test crossmatch (IAT XM). Performing an IAT XM will give a mismatched result anyway, delays the supply of blood to the patient, and provides no additional benefit or safety.STUDY DESIGN AND METHODS: From January 2002 to April 2006, the provision of blood for autoimmune hemolytic anemia (AIHA) patients with autoantibodies and no alloantibodies as well as patients with alloantibodies that exhibited a “high‐titer, low‐avidity” (HTLA) mode of reactivity was reviewed.RESULTS: A total of 222 AIHA patients (428 samples) with autoantibodies had 1585 units of red cells supplied after IAT XM; 1308 (82.5%) were mismatched. In 50 patients (80 samples) with HTLA‐like antibodies, 286 units of 328 (87.2%) were mismatched by IAT XM.CONCLUSION: No adverse reactions were reported for the study groups where “suitable” blood was provided after a serologically mismatched IAT XM. No additional benefit for these patients can be claimed by performing an IAT XM over an IS XM, as a check of ABO match. The IAT XM is both costly and time‐consuming. It is proposed that for these study group patients, a reduction to an IS XM can be applied and can be beneficial.
Two missense mutations in the CD44 gene encode two new antigens of the Indian blood group system
Authors: Poole Joyce , Tilley Louise , Warke Nicole , Spring Frances A. , Overbeeke Marijke A.M. , Van Der Mark‐Zoet José A.C.M. , Ahrens Norbert , Armstrong Diane , Williams Mark , Daniels Geoff 
Published: May 2007

BACKGROUND: Blood samples were referred over a 10‐year period from five patients whose serum samples contained antibodies to unidentified high‐incidence antigens. Three patients (A, B, C) were of Moroccan origin and their antibodies and red blood cells (RBCs) were mutually compatible, but incompatible with those of the other two patients (D, E), who were of Pakistani origin. The antibodies and RBCs of D and E were mutually compatible, but incompatible with those of Patients A, B, and C. All the antibodies were detected during pregnancy.STUDY DESIGN AND METHODS: Serologic tests, including the use of enzyme‐treated and chemically modified RBCs, suggested a relationship to CD44 (Indian blood group system). The monoclonal antibody immobilization of erythrocyte antigens (MAIEA) assay with monoclonal CD44 antibodies, immunoblotting of RBC membranes, and CD44 gene sequencing were carried out.RESULTS: Positive reactions in the MAIEA assay confirmed that the patients' antibodies are directed at CD44. Immunoblotting with two of the antibodies gave positive reactions of identical size to monoclonal anti‐CD44 and failed to react with the RBCs of a CD44‐deficient patient. One of the antibodies reacted with purified CD44. Sequencing of Exons 1 to 5 of CD44 revealed 255C>G in Exon 3 for A, B, and C encoding H85Q and 488C>A in Exon 5 for D and E encoding T163R.CONCLUSION: Two novel CD44 antigens of high incidence have been identified: IN3 (INFI) and IN4 (INJA) in the IN (Indian) blood group system. Lack of IN3 and IN4 results from homozygosity for mutations encoding H85Q and T163R in CD44, respectively.
Shock and Awe vs. Hearts and Minds at the Movies
Authors: GARDELS NATHAN , MEDAVOY MIKE 
Published: May 2007

In the age of globalization we are no longer home alone. Migration brings other worlds into our own just as the global reach of the mass media transmits our world into the hearts and minds of others. The resulting clashes and fusions are shaping the soul of the first global civilization in the making.In this section, one of Hollywood's top producers, Mike Medavoy, and the Oscar‐nominated director of Babel, Alejandro González Iñárritu share their thoughts with NPQ.
North Korea Accord Is a Model for Iranian Crisis
Authors: BLIX HANS 
Published: May 2007

As the Soviet Union collapsed and China shifted to the market, it was said that communism was nothing more than the longest road between capitalism and capitalism. It now looks like the Bush‐era neo‐conservatism is similarly a long road back to the realism from whence it departed. The US is now talking with North Korea and Iran instead of preempting them through war.A dissident Iranian scientist, the former top UN arms inspector and a former US secretary of state ponder this state of affairs.
The Cost of the Last Six Years, From North Korea to Kosovo
Authors: ALBRIGHT MADELEINE 
Published: May 2007

As the Soviet Union collapsed and China shifted to the market, it was said that communism was nothing more than the longest road between capitalism and capitalism. It now looks like the Bush‐era neo‐conservatism is similarly a long road back to the realism from whence it departed. The US is now talking with North Korea and Iran instead of preempting them through war.A dissident Iranian scientist, the former top UN arms inspector and a former US secretary of state ponder this state of affairs.
The Efficacy of Inducibility and Circumferential Ablation with Pulmonary Vein Isolation in Patients with Paroxysmal Atrial Fibrillation
Authors: CHANG SHIH‐LIN , TAI CHING‐TAI , LIN YENN‐JIANG , WONGCHAROEN WANWARANG , LO LI‐WEI , TUAN TA‐CHUAN , UDYAVAR AMEYA R. , CHANG SHENG‐HSIUNG , TSAO HSUAN‐MING , HSIEH MING‐HSIUNG , HU YU‐FENG , CHEN YI‐JEN , CHEN SHIH‐ANN 
Published: May 2007

Introduction: Some conflicting results of the efficacy of the inducibility test used in the catheter ablation of atrial fibrillation (AF) have been reported. The aim of this study was to investigate the inducibility and efficacy of circumferential ablation with pulmonary vein isolation (PVI) in patients with paroxysmal AF and its relationship to the atrial substrate. Methods and Results: This study consisted of 88 patients with paroxysmal AF who underwent catheter ablation. Electroanatomic mapping using a NavX system was performed and the biatrial voltage was obtained during sinus rhythm. After successful circumferential ablation with PVI, an inducibility test was performed to determine the requirement for creating left atrial (LA) ablation line. After procedure, patients with inducible AF had a higher recurrence rate than did those with noninducibility of AF (55% vs 18%, P = 0.02). The patients with inducible AF after the PVI had a lower biatrial voltage than did those with negative inducibility. The patients with inducible AF after the final procedure who had a recurrence had a lower LA voltage (1.3 ± 0.4 vs 1.8 ± 0.4 mV, P = 0.02) and longer LA total activation time (99 ± 18 vs 88 ± 13 msec, P = 0.02) than did those with noninducible AF and no recurrence. None of the patients had occurrence of LA flutter during the follow‐up. Conclusion: After a single procedure of circumferential ablation with PVI and noninducibility, 82% patients did not have recurrence of AF. The inducibility of AF was related to the recurrence of AF. The atrial substrate affected the outcome of the inducibility.
Supermarkets, wholesalers, and tomato growers in Guatemala
Authors: Hernández Ricardo , Reardon Thomas , Berdegué Julio 
Published: May 2007

AbstractThe article shows the asset‐related determinants and the impacts of the participation of small farmers in supermarkets versus traditional market channels in Guatemala. Compared to farmers selling only to the traditional market channels, farmers selling to supermarket channels are larger (but are in the upper tier of the “small farmer” category), have more capital, and are much more specialized in commercial horticulture in general and in tomatoes in particular. While they have higher yields, they also have higher input use, including use of chemicals. In fact, they severely overuse pesticides and fungicides. Moreover, these greater input expenditures mean that their profit rates are roughly similar to those of farmers in the traditional market channel. Supermarket‐channel farmers prefer the more demanding wholesale‐supermarket channel because it offers lower risks and lower transaction costs to market, a variety of quality grades of tomatoes, all year long. In turn, the supermarkets, who do not buy direct but rather source from a few specialized‐dedicated wholesalers, rely on this year‐round supply, lower transaction costs, and consistency of quality.