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21.
Responses of cytotoxic T-cells (Tc) to human cytomegalovirus (CMV) represent the predominant mechanism by which hosts resist CMV infection. The CMV major immediate-early protein (IE) is present throughout the virus replicative cycle. Studies were performed to determine whether Tc specific for IE effectively lyse CMV-infected targets and are thus capable of providing protective immunity against infection. After in vitro stimulation of peripheral blood mononuclear cells with CMV-infected autologous fibroblasts, Tc specific for IE were not readily detectable in CMV-reactive polyclonal Tc lines. However, after stimulation of peripheral blood mononuclear cells with cells selectively expressing IE, weak but detectable IE-specific Tc responses were observed. The frequency of IE-specific Tc clones derived from cultures stimulated with IE-expressing cells was 50 to 100 times lower than the frequency of Tc clones specific for other CMV proteins isolated from cultures stimulated with CMV-infected cells. All of the IE-specific Tc clones, which efficiently lysed targets selectively expressing IE, demonstrated minimal lysis of CMV-infected fibroblasts, despite abundant IE expression in these target cells. In contrast to these results with IE, other viral proteins were efficiently presented during all phases of CMV infection. These data suggest that CMV has evolved a unique mechanism for selectively limiting the presentation of the potentially immunogenic IE protein, which may preclude IE-specific Tc from providing protective immunity to CMV infection. 相似文献
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The peripherin gene has three potential ATG translation initiation sites at positions 38, 56, and 290. The second ATG has been proposed to be the initiation codon used for translation of the protein, but there is no experimental evidence for this conjecture. We have isolated a full-length peripherin cDNA (designated as p61-11) from a rat brain cDNA library. Upon sequencing, we found that this cDNA contains a point mutation at the second potential translation initiation codon, which changes this ATG to ACG. When expressed in SW13 cl.2 vim- cells, a cell line without any detectable cytoplasmic intermediate filaments, the protein product of p61-11 cannot form a filamentous network and the major product is 45 kDa in size, which is most likely initiated from the third ATG. The protein product from the first ATG (57 kDa in size) of p61-11 is also detected albeit in smaller amounts. We introduced a frame-shift mutation upstream of the third ATG in p61-11 to create p61-11FS and showed that the third ATG is able to initiate translation efficiently even in the presence of the first ATG, and the 45 kDa protein leads to a diffuse nonfilamentous staining pattern in vim- cells confirming that the first ATG may not be the preferred translation initiation codon, since it cannot suppress a downstream ATG. We increased the translation efficiency from the first ATG of p61-11 by mutating the three nucleotides preceding this first ATG and thereby placing it in a better Kozak consensus sequence for translation initiation. The resulting 57 kDa protein is able to form a filamentous network in vim- cells. We corrected the mutation in the original p61-11 by polymerase chain reaction and generated two peripherin constructs: perM1M2 (which contains all three translation initiation codons) and per delta 1M2 (the first ATG is deleted, but the other two are present). When transfected, their protein products, about 57 kDa in size, form filamentous networks in the absence of other cytoplasmic intermediate filaments. Since there is no 45 kDa protein detected for these latter two constructs, it is reasonable to conclude that in the presence of the second ATG, little or no translation is initiated from the third ATG. Taken together, these results strongly suggest that the second ATG is the preferred translation initiation codon for the peripherin gene. 相似文献
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The annual time-series analysis examines the impact of changes in per capita alcohol consumption (NIAAA,AEDS) on changes in community hospital admission rates (AHA) in the United States from 1950 to 1992 (n = 43). Increases in per capita alcohol consumption were expected to increase hospital admission rates contemporaneously and several years thereafter following an exponential risk function. Distributed lag models based on differenced data controlling for changes in: (1) per capita cigarette consumption; (2) private hospital insurance coverage; (3) the drinking age population; (4) per capita disposable personal income; and (5) health care regulatory interventions show a contemporaneous effect of per capita alcohol consumption on hospital admission rates. The time-series analyses imply that between 22-26% of US community hospital admissions are alcohol related. A comparable analysis indicates that per capita alcohol and tobacco expenditures contribute to approximately 28% of US community hospital admissions. The absence of statistically significant lagged effects is inconsistent with an exponentially declining risk functions. However, the contemporaneous effects of per capita alcohol and tobacco consumption suggest that a reduction in smoking and drinking will produce quick reductions in morbidity and hospitalizations. 相似文献
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Solanum melongena (eggplant) cv. Picentia and the wild species Solanum integrifolium were transformed with both a wild type (wt) and four mutagenized versions of Bacillus thuringiensis (Bt) gene Bt43 belonging to the cry3 class. The Bt gene was partly modified in its nucleotide sequence by replacing four target regions (W: +1 to +170; X: +592 to +1057; Y: +1203 to +1376; Z: +1376 to +1984) with synthetic fragments obtained by polymerase chain reaction amplification of crude oligonucleotides. The synthetic Bt genes were designed to avoid, in their modified regions, sequences such as ATTTA sequence, polyadenylation sequences and splicing sites, which might destabilize the messenger RNA. Furthermore, the codon usage was improved for a better expression in the plant system. The amino acid composition was not altered. Four versions of the modified Bt gene were obtained, BtE, BtF, BtH and BtI, with a nucleotide subtitution percentage of 8.2, 8.6, 14, and 16%, respectively, in comparison to the wt gene Bt43. Modified versions contained different subsets of substituted regions: BtE-W + Z, BtF - Y + Z, BtH-X + Y + Z, BtI - W + X + Y + Z. In the final modified version (BtI), overall guanine+cytosine was increased from the 34.1% of the wt gene to 45.5%, and most of the destabilizing sequences were eliminated. Transgenic plants obtained with the more modified versions, BtH and BtI, were fully resistant to Leptinotarsa decemlineata Say first- and third-instar larvae, while Bt43 wt, BtE and BtF genotypes did not cause mortality and did not affect larval development. 相似文献
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This study examined the relationship of several maternal variables to the duration of exclusive breast-feeding and the total duration of breast-feeding, along with attitudes, perceptions, and beliefs about breast-feeding among women living in poor neighborhoods of Managua, Nicaragua. The field work was carried out in December 1992 and January 1993 using qualitative and quantitative methods. A structured questionnaire was administered in interviews with 556 mothers of children under 12 months of age, and meetings of four directed discussion groups were held, in which a total of 20 women participated. At one week of age, almost all the children of the mothers who were surveyed had been breast-fed, but only 45% had been exclusively breast-fed. At 12 weeks old, 30% were already completely weaned. The discussion groups revealed the coexistence of positive opinions about both breast-feeding and bottle-feeding. However, exclusive breast-feeding was considered harmful for the mother, and breast milk was not thought to be sufficient nourishment for the child. Previous experience was strongly related to the duration of exclusive breast-feeding and to total breast-feeding duration. Attitudes, social support, and work situation were important factors influencing the total length of time women breast-fed a child. In general, the results obtained through the interviews and in the discussion groups were in agreement and showed that the elements needed to promote exclusive breast-feeding were social support, a favorable community environment, and policies that dealt with problems faced by working mothers. The complementary research methods were useful for obtaining information about the relative importance of different factors that determine the duration of breast-feeding and for understanding that practice in greater depth from the mother's point of view. 相似文献
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