RESUMO
The combination of supported rhodium metal catalysts and supercritical carbon dioxide solvent was effective for the stereoselective ring hydrogenations of aromatic compounds at low temperature. Higher solubility of hydrogen in supercritical carbon dioxide provides higher concentration of hydrogen on the rhodium surface, but lower that of the intermediate on rhodium surface, which suppresses the flipping of surface intermediate, leading to higher catalyst activities and cis selectivities to the corresponding ring-hydrogenated products as compared with those in organic solvents.
RESUMO
In blunt force trauma to the head caused by attacks with blunt instruments, contact forces can be estimated based on the conservation of momentum if impact velocities are known. The aims of this work were to measure maximum striking velocities and to examine the influence of rod parameters such as rod mass and length as well as volunteer parameters such as sex, age, body height, body mass, body mass index and the average amount of physical exercise. Steel rods with masses of 500, 1000 and 1500 g as well as lengths of 40, 65 and 90 cm were exemplarily tested as blunt instruments. Twenty-nine men and 22 women participated in this study. Each volunteer performed several vertical strikes with the steel rods onto a passive immobile target. Maximum striking velocities were measured by means of a Qualisys motion capture system using high-speed cameras and infrared light. Male volunteers achieved maximum striking velocities between 14.0 and 35.5 m/s whereas female volunteers achieved values between 10.4 and 28.3 m/s. Results show that maximum striking velocities increased with smaller rod masses and less consistently with higher rod lengths. Statistically significant influences were found in the volunteers' sex and average amount of physical exercise.
Assuntos
Desenho de Equipamento , Cinética , Armas , Ferimentos não Penetrantes , Feminino , Ciências Forenses , Humanos , Modelos Lineares , Masculino , Aptidão Física , Fatores SexuaisRESUMO
Forensic experts often have to assess injury and fatality risks in the context of violent blunt force trauma. Maximum striking velocities in one- and two-handed strikes with a rod-like implement can be of particular interest. Current literature lacks studies addressing this problem. The purpose of this study was therefore to measure and analyse maximum striking velocities in one-handed and two-handed strikes in female and male volunteers. We hypothesised higher striking velocities in two-handed strikes compared to one-handed strikes. Fifty volunteers performed one- and two-handed strikes from top to bottom using a steel rod of 65 cm length and 1000 g weight. A Qualisys™ Motion Analysis system registered displacements of reflecting markers fixed to the rod as well as to the volunteer's body. In one-handed strikes, the mean maximum striking velocity was 17.2 m/s in the female sample and 23.9 m/s in the male sample. Statistically not significantly different maximum striking velocities were found in two-handed strikes with mean values of 18.3 m/s in the female sample and 24.2 m/s in the male sample. Female and male volunteers also yielded similar mean maximum striking velocities in two-handed strikes comparing 'overhead' and 'overshoulder' striking techniques. In conclusion, the striking technique did not relevantly influence maximum striking velocities in our setup.
Assuntos
Desenho de Equipamento , Armas , Ferimentos não Penetrantes , Fenômenos Biofísicos/fisiologia , Feminino , Ciências Forenses , Mãos , Humanos , Cinética , Masculino , Aptidão Física , Fatores Sexuais , SuéciaRESUMO
As the availability of data sets increases, meta-analysis leveraging aggregated and interoperable data types is proving valuable. This study leveraged a meta-analysis workflow to identify mutations that could improve robustness to reactive oxygen species (ROS) stresses using an industrially important melatonin production strain as an example. ROS stresses often occur during cultivation and negatively affect strain performance. Cellular response to ROS is also linked to the SOS response and resistance to pH fluctuations, which is important to strain robustness in large-scale biomanufacturing. This work integrated more than 7000 E. coli adaptive laboratory evolution (ALE) mutations across 59 experiments to statistically associate mutated genes to 2 ROS tolerance ALE conditions from 72 unique conditions. Mutant oxyR, fur, iscR, and ygfZ were significantly associated and hypothesized to contribute fitness in ROS stress. Across these genes, 259 total mutations were inspected in conjunction with transcriptomics from 46 iModulon experiments. Ten mutations were chosen for reintroduction based on mutation clustering and coinciding transcriptional changes as evidence of fitness impact. Strains with mutations reintroduced into oxyR, fur, iscR, and ygfZ exhibited increased tolerance to H2O2 and acid stress and reduced SOS response, all of which are related to ROS. Additionally, new evidence was generated toward understanding the function of ygfZ, an uncharacterized gene. This meta-analysis approach utilized aggregated and interoperable multiomics data sets to identify mutations conferring industrially relevant phenotypes with the least drawbacks, describing an approach for data-driven strain engineering to optimize microbial cell factories.
Assuntos
Escherichia coli , Mutação , Estresse Oxidativo , Espécies Reativas de Oxigênio , Estresse Oxidativo/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Melatonina/metabolismo , Evolução Molecular Direcionada/métodosRESUMO
Leg morphology is an important outcome of evolution. A remarkable morphological leg feature is the existence of biarticular muscles that span adjacent joints. Diverse studies from different fields of research suggest a less coherent understanding of the muscles' functionality in cyclic, sagittal plane locomotion. We structured this review of biarticular muscle function by reflecting biomechanical template models, human experiments and robotic system designs. Within these approaches, we surveyed the contribution of biarticular muscles to the locomotor subfunctions (stance, balance and swing). While mono- and biarticular muscles do not show physiological differences, the reviewed studies provide evidence for complementary and locomotor subfunction-specific contributions of mono- and biarticular muscles. In stance, biarticular muscles coordinate joint movements, improve economy (e.g. by transferring energy) and secure the zig-zag configuration of the leg against joint overextension. These commonly known functions are extended by an explicit role of biarticular muscles in controlling the angular momentum for balance and swing. Human-like leg arrangement and intrinsic (compliant) properties of biarticular structures improve the controllability and energy efficiency of legged robots and assistive devices. Future interdisciplinary research on biarticular muscles should address their role for sensing and control as well as non-cyclic and/or non-sagittal motions, and non-static moment arms.
Assuntos
Robótica , Braço , Fenômenos Biomecânicos , Humanos , Perna (Membro) , Músculo EsqueléticoRESUMO
The 4,639,221-base pair sequence of Escherichia coli K-12 is presented. Of 4288 protein-coding genes annotated, 38 percent have no attributed function. Comparison with five other sequenced microbes reveals ubiquitous as well as narrowly distributed gene families; many families of similar genes within E. coli are also evident. The largest family of paralogous proteins contains 80 ABC transporters. The genome as a whole is strikingly organized with respect to the local direction of replication; guanines, oligonucleotides possibly related to replication and recombination, and most genes are so oriented. The genome also contains insertion sequence (IS) elements, phage remnants, and many other patches of unusual composition indicating genome plasticity through horizontal transfer.
Assuntos
Escherichia coli/genética , Genoma Bacteriano , Análise de Sequência de DNA , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Bacteriófago lambda/genética , Composição de Bases , Sítios de Ligação , Mapeamento Cromossômico , Replicação do DNA , Elementos de DNA Transponíveis , DNA Bacteriano/genética , Genes Bacterianos , Dados de Sequência Molecular , Mutação , Óperon , RNA Bacteriano/genética , RNA de Transferência/genética , Recombinação Genética , Sequências Reguladoras de Ácido Nucleico , Sequências Repetitivas de Ácido Nucleico , Homologia de Sequência de AminoácidosRESUMO
Our ability of screening broad communities for clinically asymptomatic diseases critically drives population health. Sensory chewing gums are presented targeting the tongue as 24/7 detector allowing diagnosis by "anyone, anywhere, anytime". The chewing gum contains peptide sensors consisting of a protease cleavable linker in between a bitter substance and a microparticle. Matrix metalloproteinases in the oral cavity, as upregulated in peri-implant disease, specifically target the protease cleavable linker while chewing the gum, thereby generating bitterness for detection by the tongue. The peptide sensors prove significant success in discriminating saliva collected from patients with peri-implant disease versus clinically asymptomatic volunteers. Superior outcome is demonstrated over commercially available protease-based tests in saliva. "Anyone, anywhere, anytime" diagnostics are within reach for oral inflammation. Expanding this platform technology to other diseases in the future features this diagnostic as a massive screening tool potentially maximizing impact on population health.Early detection of gum inflammation caused by dental implants helps prevent tissue damage. Here, the authors present a peptide sensor that generates a bitter taste when cleaved by proteases present in peri-implant disease, embed it in a chewing gum, and compare the probe to existing sensors using patient saliva.
Assuntos
Goma de Mascar , Implantes Dentários , Gengivite/diagnóstico , Metaloproteinases da Matriz/metabolismo , Peptídeos/metabolismo , Periodontite/diagnóstico , Paladar , Gengivite/metabolismo , Humanos , Periodontite/metabolismo , Saliva/enzimologiaRESUMO
The preparation method of metal based Indian traditional drugs involves conversion of a pure metal into its oxide by repeated high temperature calcination cycles. In this work, the effect of number of calcination cycles followed in the preparation of tin oxide based Ayurvedic drug, 'vanga bhasma' was studied by a systematic characterization of the drug samples after various calcination stages. It was found that tin was in the form of Sn4+ state and that the formation of SnO2 proceeded step-wise through Sn(OH)4.
Assuntos
Química Farmacêutica/métodos , Ayurveda , Extratos Vegetais/química , Compostos de Estanho/química , Oxirredução , Tamanho da PartículaRESUMO
bcl-2 is the first member of a new class of protooncogenes the products of which inhibit programmed cell death (PCD) or apoptosis. We have previously determined that Bcl-2 is expressed in a significant percentage of untreated primary neuroblastoma (NBL) tumors. In these specimens Bcl-2 expression correlated with other markers of poor prognosis suggesting a role for Bcl-2 in the malignant behavior of NBL tumor cells. To investigate this possibility, a Bcl-2-negative human NBL cell line (Shep-1) was transfected with a bcl-2 expression vector (pSFFVneo-bcl-2). Multiple unique clones were isolated which showed variable levels of Bcl-2 protein by quantitative immunoprecipitation. Vector-transfected controls were generated simultaneously. Clones expressing high levels of Bcl-2 were resistant to cisplatin- and etoposide-induced cytotoxicity in a dose-dependent manner. Analysis of propidium iodide-stained nuclei by flow cytometry after cisplatin or etoposide treatment revealed marked DNA degradation in vector-transfected controls whereas bcl-2 transfectants showed a dose-dependent inhibition of DNA degradation. Analysis by pulsed-field gel electrophoresis revealed relatively large fragment DNA degradation (approximately 50 kilobases) in the absence of internucleosomal degradation in vector-transfected control cells treated with either cisplatin or etoposide. In contrast, Bcl-2-expressing cells showed significantly less DNA degradation at all time points. These single gene transfection experiments have revealed that expression of Bcl-2 renders specific NBL cells resistant to chemotherapy-induced PCD and support the hypothesis that Bcl-2 enhances the malignant phenotype of NBL by promoting tumor resistance to chemotherapy agents.
Assuntos
Apoptose/efeitos dos fármacos , Cisplatino/farmacologia , Etoposídeo/farmacologia , Neuroblastoma/tratamento farmacológico , Neuroblastoma/patologia , Proteínas Proto-Oncogênicas/fisiologia , Apoptose/fisiologia , Divisão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , DNA de Neoplasias/efeitos dos fármacos , DNA de Neoplasias/metabolismo , Resistência a Medicamentos , Citometria de Fluxo , Expressão Gênica , Humanos , Neuroblastoma/genética , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas c-bcl-2 , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
bcl-x is a member of the bcl-2 family of genes and by alternative splicing gives rise to two distinct mRNAs: bcl-xL and bcl-xS. We have previously investigated the expression of Bcl-x in neuroblastoma (NB) cell lines and have shown that Bcl-xL is expressed and functions to inhibit chemotherapy-induced apoptosis. However, none of the NB cell lines expressed Bcl-xS. The aim of the present study was to determine the effects of Bcl-xS expression on the viability of NB cells. A panel of NB cell lines (CHP-382, GOTO, SHEP-1, SHSY-5Y, and GI-CA-N) were infected with either a bcl-xS adenovirus (pAdRSV-bcl-xS) or a control virus (pAdRSV-lac-z). NB cells showed loss of viability with both viruses, although the bcl-xS virus was most toxic. Importantly, infection with the bcl-xS adenovirus resulted in rapid loss of cell viability, DNA fragmentation, and morphological features of apoptosis even in NB cells transfected to overexpress Bcl-2 and Bcl-xL. These findings suggest that deregulated expression of Bcl-xS using an adenovirus may provide a novel mechanism for initiating cell death in tumors that express Bcl-2 or Bcl-xL.
Assuntos
Apoptose/genética , Genes bcl-2/genética , Neuroblastoma/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2 , Proteínas Proto-Oncogênicas/genética , Adenoviridae/genética , Fragmentação do DNA , DNA de Neoplasias/genética , Vetores Genéticos/genética , Humanos , Neuroblastoma/genética , Neuroblastoma/patologia , Proteínas Proto-Oncogênicas/metabolismo , Transfecção , Células Tumorais Cultivadas , Proteína bcl-XRESUMO
The copper based Indian traditional drug 'tamra bhasma' is administered for various ailments since long. Its synthesis involves treating metallic copper with plant juices and then repeated calcination in presence of air so that the metallic state is transformed into the corresponding oxide form traditionally known as 'bhasma'. In this work, we present a systematic characterization of this traditional drug using various techniques like X-ray diffraction (XRD), scanning electron microscopy (SEM)-energy dispersive X-ray analysis (EDX), X-ray photoelectron spectroscopy (XPS), infrared spectroscopy (IR), thermogravimetry (TG) and surface area measurement. The results obtained were found to match very well with those of a standard copper oxide confirming the composition of the drug sample. In addition, some specific findings were also made which could help in interpreting the therapeutic properties of the traditional drug 'tamra bhasma'.
Assuntos
Ácido Ascórbico/análise , Cobre/análise , Compostos Ferrosos/análise , Combinação de Medicamentos , Composição de Medicamentos , Índia , Ayurveda , Microscopia Eletrônica de Varredura , Tamanho da Partícula , Espectrometria por Raios X , Espectrofotometria Infravermelho , Propriedades de Superfície , Termogravimetria , Difração de Raios XRESUMO
Remission marrow from patients with BCR-ABL+ acute lymphoblastic leukemia (ALL) achieving clinical remission (CR) after induction or consolidation chemotherapy according to the German multicenter adult ALL (GMALL) protocol showed high titers of residual BCR-ABL+ cells. Therefore, we initiated a pilot study to monitor circulating BCR-ABL+ cells and to collect, purge, and autograft peripheral blood stem cells (PBSC) in these patients. After GMALL 05/93 high-risk phase II of induction chemotherapy (high-dose AraC 3 g/m2 x 8 does and mitoxantrone 10 mg/m2 x 3 doses), patients received 5-10 micrograms/kg subcutaneous recombinant human granulocyte colony-stimulating factor (rhG-CSF) daily. Mobilized CD34+ cells peaked between 20 and 26 days after starting chemotherapy at 4.8-75.6 (median 10.8) x 10(4)/mL peripheral blood (PB) (n = 5). Patients treated with additional chemotherapy cycles failed to mobilize adequate numbers of CD34+ cells. PB stem cells (PBSC) were purged using a cocktail of CD10, CD19, and AB4 monoclonal antibodies (mAbs) coupled to immunomagnetic beads (IMB). The median recoveries of total nucleated cells (TNC) and CD34+ cells after mAb/IMB purging were 84 and 81%. The peak numbers of CD34+ cells collected in a single leukapheresis were median 8.6 x 10(6)/kg pre- and 5.2 x 10(6)/kg postpurge (n = 4). The absolute prepurge CD19+ cells were as low as median 2.7 (range 1.4-19) x 10(6) per leukapheresis. Residual BCR-ABL+ cells in unpurged leukapheresis products were assessed by limiting-log10-dilution nested reverse-transcriptase polymerase chain reaction (RT-PCR) as one in 10(5) to one in 10(6) normal cells and were consistently undetectable in all purged PBSC autografts. We conclude that sufficient numbers of CD34+ cells for PBSCT can be collected after phase II but not at later stages of the GMALL 05/93 high risk protocol; PBSC grafts are 3 log less contaminated with residual BCR-ABL+ cells compared to an historical series of 13 autologous BM grafts; and purging of PBSC with mAb/IMB is feasible with minor loss of CD34+ cells and abolished BCR-ABL signals in the grafts.
Assuntos
DNA/análise , Proteínas de Fusão bcr-abl/genética , Células-Tronco Hematopoéticas/citologia , Separação Imunomagnética , Cromossomo Filadélfia , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Adulto , Antígenos CD19/análise , Antígenos CD34/análise , Feminino , Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas/química , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Reação em Cadeia da Polimerase , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Fatores de Tempo , Transplante AutólogoRESUMO
Electrophoretic separation of macrorestriction fragments containing a particular genomic interval has until recently depended on fortuitously placed native rare restriction sites. We present new IS10-based transposons carrying the yeast intron-encoded I-SceI restriction site which is absent from most prokaryotic and eukaryotic genomes. Construction of the plasmid vectors containing them is described. Analysis by conventional or Pulsed Field gel electrophoresis of the DNA fragments generated by the I-SceI digestion reveals the physical distance between genomic insertions of these transposons: use of the same approach to subdivide the chromosome of Escherichia coli K-12 into equivalently sized contiguous/nonoverlapping I-SceI fragments is demonstrated. Because coordinates for the loci delimited by their insertions can be readily determined in different isolates by either physical or genetic manipulations, these transposons allow sufficient flexibility for species-wide bacterial genomics.
Assuntos
Elementos de DNA Transponíveis , Desoxirribonucleases de Sítio Específico do Tipo II/metabolismo , Endodesoxirribonucleases/metabolismo , Genoma Bacteriano , Animais , Sítios de Ligação , Escherichia coli/genética , Bactérias Gram-Negativas/genética , Proteínas de Saccharomyces cerevisiaeRESUMO
Genetic and biophysical techniques have traditionally been applied to genome mapping independently of one another. We present a series of Escherichia coli mini-Tn10 insertions that contain the rare-cutting polylinker 1 (RCP1) of rare restriction sites [including BlnI/AvrII, SpeI, NheI, XbaI, NotI, PacI and SfiI; Mahillon and Kleckner, Gene 116 (1992) 69-74] which allows them to be used not just for genetic mapping, but also for rapid physical mapping and integrated physical and genetic mapping of the E. coli chromosome. Their isolation and their physical and genetic coordinates in K-12 strain MG1655 are presented. Also, their use in purifying insertion-delimited DNAs from E. coli K-12 and in macrorestriction mapping of a pathogenic strain's chromosome is demonstrated. These insertions allow integration of (i) different macrorestriction patterns of a single strain's chromosome, (ii) the physical map of a single strain's chromosome with the genetic map of the species, and (iii) the physical maps of different strains' chromosomes.
Assuntos
Cromossomos Bacterianos/ultraestrutura , DNA Bacteriano , Escherichia coli/genética , Mapeamento por Restrição , Mapeamento Cromossômico , Elementos de DNA Transponíveis , Desoxirribonucleases de Sítio Específico do Tipo II , Genes BacterianosRESUMO
A transposon-based method of introducing unique restriction sites was used for subdivision of the Escherichia coli genome into a contiguous series of large non-overlapping segments spanning 2.5Mb. The segments, sizes ranging from 150 to 250kb, were isolated from the chromosome using the inserted restriction sites and shotgun cloned into an M13 vector for DNA sequencing. These shotgun sizes proved easily manageable, allowing the genomic sequence of E. coli to be completed more efficiently and rapidly than was possible by previously available methods. The 9bp duplication generated during transposition was used as a tag for accurate splicing of the segments; no further sequence redundancy at the junction sites was needed. The system is applicable to larger genomes even if they are not already well-characterized. We present the technology for segment sequencing, results of applying this method to E. coli, and the sequences of the transposon cassettes.
Assuntos
Cromossomos Bacterianos , Elementos de DNA Transponíveis , Desoxirribonucleases de Sítio Específico do Tipo II/genética , Escherichia coli/genética , Análise de Sequência de DNA/métodos , Biblioteca Gênica , Genoma Bacteriano , Proteínas de Saccharomyces cerevisiaeRESUMO
Cerebral lateralization in females is probably modulated by the menstrual cycle such that a part of hemispheric asymmetries are diminished with an increase of the steroids estrogen and progesterone during the follicular and luteal phase and enhanced with steroid decreases during the menstrual phase. However, previous data were contradictory with regard to the hemispheric side of modulation and could not analyze which steroid is mainly responsible for cycle dependent lateralization shifts. Therefore, in the present study estrogen and progesterone changes were assessed separately and related to changes in cerebral asymmetry. Plasma levels of estrogen and progesterone were measured once during luteal and once during menstrual cycle phase while 20 females subjects performed a verbal and a figural lateralized matching task. The results showed a significant cycle phase x lateralization interaction for the right hemisphere dominated figural comparison task but not for the left hemispheric lexical condition. Although the lateralization was modulated by the menstrual cycle, a within-subject regression analysis demonstrated that the asymmetry shift was not under direct influence of estrogen or progesterone. Thus, the present study provides further empirical support for cycle dependent alterations in lateralization but makes it unlikely that this effect is directly caused by estrogen or progesterone plasma level variations.
Assuntos
Encéfalo/fisiologia , Estrogênios/sangue , Lateralidade Funcional/fisiologia , Ciclo Menstrual/fisiologia , Progesterona/sangue , Adulto , Feminino , Humanos , Testes de Linguagem , Fase Luteal/fisiologia , Tempo de Reação , Percepção da FalaRESUMO
When given a choice between two otherwise equivalent options - one in which the probability information is stated and another in which it is missing - most people avoid the option with missing probability information (Camerer & Weber, 1992). This robust, frequently replicated tendency is known as the ambiguity effect. It is unclear, however, why the ambiguity effect occurs. Experiments 1 and 2, which separated effects of the comparison process from those related to missing probability information, demonstrate that the ambiguity effect is elicited by missing probabilities rather than by comparison of options. Experiments 3 and 4 test predictions drawn from the literature on behavioral ecology. It is suggested that choices between two options should reflect three parameters: (1) the need of the organism, (2) the mean expected outcome of each option; and (3) the variance associated with each option's outcome. It is hypothesized that unknown probabilities are avoided because they co-occur with high outcome variability. In Experiment 3 it was found that subjects systematically avoid options with high outcome variability regardless of whether probabilities are explicitly stated or not. In Experiment 4, we reversed the ambiguity effect: when participants' need was greater than the known option's expected mean outcome, subjects preferred the ambiguous (high variance) option. From these experiments we conclude that people do not generally avoid ambiguous options. Instead, they take into account expected outcome, outcome variability, and their need in order to arrive at a decision that is most likely to satisfy this need.
Assuntos
Aprendizagem da Esquiva , Comportamento de Escolha , Dissonância Cognitiva , Assunção de Riscos , Adulto , Análise de Variância , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Modelos PsicológicosRESUMO
Residual leukemia was evaluated in autologous bone marrow grafts harvested in first (n = 11) or second (n = 3) complete remission from 14 patients with BCR-ABL-positive acute lymphoblastic leukemia after treatment according to the German multicenter ALL protocols. The intervals from diagnosis to BM harvest were median 159 (range 78-463) and from preceding chemotherapy to BM harvest median 39 (range 26-69) days, respectively. A limiting log(10)-dilution RT-PCR was used to semiquantify BCR-ABL-positive cells. All autografts appeared to be significantly contaminated with residual leukemic cells. The BCR-ABL-specific titers ranged from 1:10(3) to 1:10(6) (median 1:10(4)) above the limit of detection. This was the rationale to purge the grafts using two cycles of IgM anti-CD10, CD19, and AB4 MoAbs-coated immunomagnetic beads (IMB). Purging depleted median 3 (range 2-4) logs of residual leukemia, resulting in a median 1:10(1) (range 1:10(0) to 1:10(3)) postpurge BCR-ABL-specific titer. The second purging cycle accounted for 1 log of depletion. The mean +/- s.e.m. post-purge recoveries of MNC and CFU-GM were 59 +/- 4%, and 61 +/- 9%, respectively. We conclude that all BCR-ABL-positive ALL patients achieving CR by cytological criteria have critically high levels of residual leukemia in their bone marrow, which can be reduced by median 3 log using immunomagnetic bead purging.
Assuntos
Purging da Medula Óssea , Transplante de Medula Óssea , Proteínas de Fusão bcr-abl/análise , Separação Imunomagnética , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Adolescente , Adulto , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasia Residual , Reação em Cadeia da PolimeraseRESUMO
The most virulent newborn meningitis-associated Escherichia coli are of the serotype O18:K1:H7. We previously isolated a large number of E. coli O18:K1:H7 mutants resulting from transposon TnphoA mutagenesis that fail to invade brain microvascular endothelial cells. We have now determined Ic locations of 45 independent insertions. Twelve were localized to the 98 min region, containing a 120 kb segment that is characteristic of E. coli O18:K1:H7. Another, the previously described insertion ibe-10::TnphoA, was localized to the 87 min region, containing a 20 kb segment found in this E. coli. These noninvasion mutations may define new O18:K1:H7 pathogenicity islands carrying genes for penetration of the blood-brain barrier of newborn mammals.
Assuntos
Cromossomos Bacterianos/genética , Elementos de DNA Transponíveis/genética , Escherichia coli/genética , Genoma Bacteriano , Mutagênese Insercional , Fosfatase Alcalina/genética , Proteínas de Bactérias/genética , Barreira Hematoencefálica , Células Cultivadas , Mapeamento Cromossômico , Endotélio Vascular/microbiologia , Escherichia coli/classificação , Escherichia coli/isolamento & purificação , Escherichia coli/patogenicidade , Infecções por Escherichia coli/microbiologia , Humanos , Recém-Nascido , Meningites Bacterianas/microbiologia , Proteínas Recombinantes de Fusão/genética , Sorotipagem , Virulência/genéticaRESUMO
Pulsed-field gel electrophoresis (PFGE), because of the increased sensitivity it affords over other methods of bacterial genotyping, represents a potentially powerful tool for the characterisation of isolates from hospital infections. Genomic fingerprinting by PFGE was applied to all clinical isolates of Escherichia coli obtained from blood during a 6-month period (78 isolates, 58 patients) at the University of Michigan Medical Center. The rare-restriction patterns of these isolates, in contrast to those of isolates from the E. coli reference collection (ECOR), were not randomly distributed through the E. coli species. Four related clusters, which represented c. 21% of the blood isolates, were identified. Two of these genotypic clusters were also clustered temporally, their members all being isolated within the same 2-week period, while the other two clusters spanned the study period. These observations indicate in-hospital endemic vectors or the occurrence of specialised E. coli lineages that are capable of invading the bloodstream and exploiting in-hospital vectors, or both.