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41.
Johanna H Kattenberg Eleanor A Ochodo Kimberly R Boer Henk DFH Schallig Petra F Mens Mariska MG Leeflang 《Malaria journal》2011,10(1):1-18
Background
To prepare field sites for malaria vaccine trials, it is important to determine baseline antibody and T cell responses to candidate malaria vaccine antigens. Assessing T cell responses is especially challenging, given genetic restriction, low responses observed in endemic areas, their variability over time, potential suppression by parasitaemia and the intrinsic variability of the assays.Methods
In Part A of this study, antibody titres were measured in adults from urban and rural communities in Ghana to recombinant Plasmodium falciparum CSP, SSP2/TRAP, LSA1, EXP1, MSP1, MSP3 and EBA175 by ELISA, and to sporozoites and infected erythrocytes by IFA. Positive ELISA responses were determined using two methods. T cell responses to defined CD8 or CD4 T cell epitopes from CSP, SSP2/TRAP, LSA1 and EXP1 were measured by ex vivo IFN-γ ELISpot assays using HLA-matched Class I- and DR-restricted synthetic peptides. In Part B, the reproducibility of the ELISpot assay to CSP and AMA1 was measured by repeating assays of individual samples using peptide pools and low, medium or high stringency criteria for defining positive responses, and by comparing samples collected two weeks apart.Results
In Part A, positive antibody responses varied widely from 17%-100%, according to the antigen and statistical method, with blood stage antigens showing more frequent and higher magnitude responses. ELISA titres were higher in rural subjects, while IFA titres and the frequencies and magnitudes of ex vivo ELISpot activities were similar in both communities. DR-restricted peptides showed stronger responses than Class I-restricted peptides. In Part B, the most stringent statistical criteria gave the fewest, and the least stringent the most positive responses, with reproducibility slightly higher using the least stringent method when assays were repeated. Results varied significantly between the two-week time-points for many participants.Conclusions
All participants were positive for at least one malaria protein by ELISA, with results dependent on the criteria for positivity. Likewise, ELISpot responses varied among participants, but were relatively reproducible by the three methods tested, especially the least stringent, when assays were repeated. However, results often differed between samples taken two weeks apart, indicating significant biological variability over short intervals. 相似文献42.
MG Oliveira Alves CFL Carta M-E Padín-Iruegas M Pérez-Sayáns JM Suarez-Peñaranda JS Issa 《Biotechnic & histochemistry》2016,91(4):263-268
We investigated the gene and protein expressions of V-type ATPase protein subunit C1 (ATP6V1C1) in cases of oral squamous cell carcinoma (OSCC) and contralateral normal mucosa in smokers, nonsmokers and former smokers. Subjects were separated into five groups of 15: group 1, smokers with OSCC; group 2, normal contralateral mucosa of OSCC patients; group 3, chronic smokers; group 4, former smokers who had stopped smoking 1 year earlier; group 5, individuals who had never smoked. Exfoliative cytology specimens from oral mucosa of smokers, former smokers and nonsmokers showed normal gene and protein expression. We found significantly greater gene expression in the OSCC group than in the nonsmoker groups. No difference in gene expression was observed between normal contralateral mucosa and nonsmoker groups, smoker and nonsmoker groups or former smoker and nonsmoker groups. We observed intense immunostaining for ATP6V1C1 protein in all cases of OSCC and weak or no staining in smoker, former smoker and nonsmoker groups. Significantly greater expression of ATP6V1C1 protein was observed in the OSCC group compared to the other groups, which supports the role of ATP6V1C1 in effecting changes associated with oral cancer. Analysis of the mucosae of chronic smokers, former smokers and the normal contralateral mucosa of patients with OSCC showed unaltered ATP6V1C1 gene and protein expression. Early stages of carcinogenesis, represented by altered epithelium of chronic smokers, had neither gene nor protein alterations as seen in OSCC. Therefore, we infer that the changes in ATP6V1C1 occur during later stages of carcinogenesis. Our preliminary study provides a basis for future studies of using ATP6V1C1 levels for detecting early stage OSCC. 相似文献
43.
Equine arteritis virus (EAV) induces apoptosis in infected cells. Cell death caused by EAV has been studied mainly using three cell lines, BHK-21, RK-13 and Vero cells. The mechanism of apoptosis varies among cell lines and results cannot be correlated owing to differences in EAV strains used. We evaluated different markers for apoptosis in BHK-21, RK-13 and Vero cell lines using the Bucyrus EAV reference strain. Acridine orange/ethidium bromide staining revealed morphological changes in infected cells, while flow cytometry indicated the extent of apoptosis. We also observed DNA fragmentation, but the DNA ladder was detected at different times post-infection depending on the cell line, i.e., 48, 72 and 96 h post-infection in RK-13, Vero and BHK-21 cells, respectively. Measurement of viral titers obtained with each cell line indicated that apoptosis causes interference with viral replication and therefore decreased viral titers. As an unequivocal marker of apoptosis, we measured the expression of caspase-3 and caspases-8 and -9 as extrinsic and intrinsic markers of apoptosis pathways, respectively. Caspase-8 in BHK-21 cells was the only protease that was not detected at any of the times assayed. We found that Bucyrus EAV strain exhibited a distinctive apoptosis pathway depending on the cell line. 相似文献
44.
杨善元 《植物生理与分子生物学学报》1989,15(1):83-87
叶绿素b由单体聚合成多聚体后吸收光谱明显红移。叶绿素b聚集体膜在光下产生150 mV的光电位,停止照光后此电位消失速度比叶绿素a聚集体膜的慢。叶绿素b聚集体吸收光能后形成相当稳定的能化态,半生命期约几分钟。当聚集体解聚时所吸收的光能又以光的形式辐射出来。叶绿素b聚集体能化态的能量可快速而有效地传给叶绿素a聚集体,以叶绿素a的延迟发光形式释放出来。 相似文献
45.
46.
Phylogenetic relationships among prokaryotic and eukaryotic catalases 总被引:13,自引:1,他引:12
Seventy-four catalase protein sequences, including 29 bacterial, 8 fungal,
7 animal, and 30 plant sequences, were compiled, and 70 were used for
phylogenetic reconstruction. The core of the resulting tree revealed
unique, separate groups of plant and animal catalases, two groups of fungal
catalases, and three groups of bacterial catalases. The only overlap of
kingdoms occurred within one branch and involved fungal and bacterial
large-subunit enzymes. The other fungal branch was closely linked to the
group of animal enzymes. Group I bacterial catalases were more closely
related to the plant enzymes and contained such diverse taxa as the
Gram-positive Listeria seeligeri, Deinocococcus radiodurans, and
gamma-proteobacteria. Group III bacterial sequences were more closely
related to fungal and animal sequences and included enzymes from a broad
range of bacteria including high- and low-GC Gram positives,
proteobacteria, and a bacteroides species. Group II was composed of
large-subunit catalases from diverse sources including Gram positives
(low-GC Bacilli and high-GC Mycobacteria), proteobacteria, and species of
the filamentous fungus Aspergillus. These data can be interpreted in terms
of two gene duplication events that produced a minimum of three catalase
gene family members that subsequently evolved in response to environmental
demands. Horizontal gene transfer may have been responsible for the group
II mixture of bacterial and fungal large-subunit catalases.
相似文献
47.
48.
Sequence variation in the guillemot (Alcidae: Cepphus) mitochondrial control region and its nuclear homolog 总被引:4,自引:0,他引:4
We describe sequence variation in the mitochondrial control region and its
nuclear homolog in three species and seven subspecies of guillemots
(Cepphus spp.). Nuclear homologs of the 5' end of the control region were
found in all individuals. Nuclear sequences were approximately 50%
divergent from their mitochondrial counterparts and formed a distinct
phylogenetic clade; the mitochondrial-nuclear introgression event must have
predated the radiation of Cepphus. As in other vertebrates, the guillemot
control region has a relatively conserved central block flanked by
hypervariable 5' and 3' ends. Mean pairwise interspecific divergence values
among control regions were lower than those in other birds. All individuals
were heteroplasmic for the number of simple tandem nucleotide repeats
(A(n)C) at the 3' end of the control region. Phylogenetic analyses suggest
that black guillemots are basal to pigeon and spectacled guillemots, but
evolutionary relationships among subspecies remain unresolved, possibly due
to incomplete lineage sorting. Describing molecular variation in nuclear
homologs of mitochondrial genes is of general interest in phylogenetics
because, if undetected, the homologs may confound interpretations of
mitochondrial phylogenies.
相似文献
49.
A phylogenetic survey using the polymerase chain reaction (PCR) has
identified four major P element subfamilies in the saltans and willistoni
species groups of Drosophila. One subfamily, containing about half of the
sequences studied, consists of elements that are very similar to the
canonical (and active) P element from D. melanogaster. Within this
subfamily, nucleotide sequence differentiation among different copies from
the same species and among elements from different species is relatively
low. This observation suggests that the canonical elements are relatively
recent additions to the genome or, less likely, are evolving slowly
relative to the other subfamilies. Elements belonging to the three
noncanonical lineages are distinct from the canonical elements and from one
another. Furthermore, there is considerably more sequence variation, on the
average, within the noncanonical subfamilies compared to the canonical
elements. Horizontal transfer and the coexistence of multiple,
independently evolving element subfamilies in the same genome may explain
the distribution of P elements in the saltans and willistoni species
groups. Such explanations are not mutually exclusive, and each may be
involved to varying degrees in the maintenance of P elements in natural
populations of Drosophila.
相似文献
50.
Pig Reticulocytes. III. Glucose permeability in naturally occurring reticulocytes and red cells from newborn piglets 总被引:1,自引:1,他引:0
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The loss of facilitated glucose transport of red cells occurring in the newborn pig was monitored in 11 density-separated cells from birth to a 4 wk of age. At birth there was a threefold increase in glucose permeability from the lightest cells to the most dense, suggesting that cells having progressively less glucose permeability are released into the circulation as gestation proceeds. Because of extraordinary stimulation of erythropoietic activity, the uppermost top fraction constituting 2-3 percent of the total cells is composed purely of reticulocytes in the growing animal. The glucose permeability of these reticulocytes which at birth has a slow but significant rate of 3.7 μmol/ml cell x min at 25 degrees C is rapidly decreased within 3-4 days to the level of reticulocytes produced in the adult in response to phenylhydrazine assault. Moreover, reticulocytes themselves discard their membrane permeability to glucose in the course of maturation to red cells. Thus, even though reticulocytes at birth are permeable to glucose, they will become red cells practically impervious to glucose within a few days. These findings suggest that the transition from a glucose- permeable fetal state to a glucose-impermeable postnatal state is brought about by two mechanisms: (a) dilution of fetal cells by glucose-impervious cells produced coincidentally with or shortly after birth; and (b) elimination of fetal cells, which have a shorter half-life, from the circulation. 相似文献