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91.
Expression of human CD81 in transgenic mice does not confer susceptibility to hepatitis C virus infection 总被引:6,自引:0,他引:6
Masciopinto F Freer G Burgio VL Levy S Galli-Stampino L Bendinelli M Houghton M Abrignani S Uematsu Y 《Virology》2002,304(2):187-196
We previously demonstrated that hepatitis C virus (HCV) binds to human CD81 through the E2 glycoprotein. Therefore, expression of the human CD81 molecule in transgenic mice was expected to provide a new tool to study HCV infection in vivo, as the chimpanzee is the only species currently available as a laboratory animal model that can be infected with HCV. We produced transgenic mice expressing the human CD81 protein in a wide variety of tissues. We confirmed binding of recombinant E2 glycoprotein to the liver tissue as well as to thymocytes and splenic lymphocytes in the transgenic mice. We inoculated chimpanzee plasma infected with HCV into these animals. None of these transgenic animals showed evidence of viral replication. Furthermore, human CD81 transgenic mice that lack expression of endogenous mouse CD81 were also resistant to HCV infection. We conclude that expression of human CD81 alone is insufficient to confer susceptibility to HCV infection in the mouse. The presence of additional possible factors for HCV infection is discussed. 相似文献
92.
Wennberg C Kozlenkov A Di Mauro S Fröhlander N Beckman L Hoylaerts MF Millán JL 《Human mutation》2002,19(3):258-267
The D allozyme of placental alkaline phosphatase (PLAP) displays enzymatic properties at variance with those of the common PLAP allozymes. We have deduced the amino acid sequence of the PLAP D allele by PCR cloning of its gene, ALPP. Two coding substitutions were found in comparison with the cDNA of the common PLAP F allele, i.e., 692C>G and 1352A>G, which translate into a P209R and E429G substitution. A single nucleotide primer extension (SNuPE) assay was developed using PCR primers that enable the amplification of a 1.9 kb PLAP fragment. Extension primers were then used on this PCR fragment to detect the 692C>G and 1352A>G substitution. The SNuPE assay on these two nucleotide substitutions enabled us to distinguish the PLAP F and D alleles from the PLAP S/I alleles. Functional studies on the D allozyme were made possible by constructing and expressing a PLAP D cDNA, i.e., [Arg209, Gly429]PLAP, into wild-type Chinese hamster ovary cells. We determined the k(cat) and K(m), of the PLAP S, F, and D allozymes using the non-physiological substrate p-nitrophenylphosphate at an optimal pH (9.8) as well as two physiological substrates, i.e., pyridoxal-5-phosphate and inorganic pyrophosphate at physiological pH (7.5). We found that the biochemical properties of the D allozyme of PLAP are significantly different from those of the common PLAP allozymes. These biochemical findings suggest that a suboptimal enzymatic function by the PLAP D allozyme may be the basis for the apparent negative selective pressure of the PLAP D allele. The development of the SNuPE assay will enable us to test the hypothesis that the PLAP D allele is subjected to intrauterine selection by examining genomic DNA from statistically informative population samples. 相似文献
93.
Claudio Marconi Mauro Marzorati Daniele Sciuto Alessandra Ferri Paolo Cerretelli 《The Journal of physiology》2005,569(2):667-675
High-altitude Tibetans undergo a pattern of adaptations to chronic hypoxia characterized, among others, by a more efficient aerobic performance compared with acclimatized lowlanders. To test whether such changes may persist upon descent to moderate altitude, oxygen uptake of 17 male Tibetan natives lifelong residents at 3500–4500 m was assessed within 1 month upon migration to 1300 m. Exercise protocols were: 5 min treadmill walking at 6 km h−1 on increasing inclines from +5 to +15% and 5 min running at 10 km h−1 on a +5% grade. The data (mean ± s.e.m. ) were compared with those obtained on Nepali lowlanders. When walking on +10, +12.5 and +15% inclines, net of Tibetans was 25.2 ± 0.7, 29.1 ± 1.1 and 31.3 ± 0.9 ml kg−1 min−1 , respectively, i.e. 8, 10 and 13% less ( P < 0.05) than that of Nepali. At the end of the heaviest load, blood lactate concentration was lower in Tibetans than in Nepali (6.0 ± 0.9 versus 8.9 ± 0.6 m m ; P < 0.05) . During running, of Tibetans was 35.1 ± 0.8 versus 39.3 ± 0.7 ml kg−1 min−1 (i.e. 11% less; P < 0.01). In conclusion, during submaximal walking and running at 1300 m, Tibetans are still characterized by lower aerobic energy expenditure than control subjects that is not accounted for by differences in mechanical power output and/or compensated for by anaerobic glycolysis. These findings indicate that chronic hypoxia induces metabolic adaptations whose underlying mechanisms still need to be elucidated, that persist for at least 1 month upon descent to moderate altitude. 相似文献
94.
Suligoi B Massi M Galli C Sciandra M Di Sora F Pezzotti P Recchia O Montella F Sinicco A Rezza G 《Journal of acquired immune deficiency syndromes (1999)》2003,32(4):424-428
We evaluated a procedure for identifying recent HIV infections, using sequential serum samples from 47 HIV-positive persons for whom the seroconversion date could be accurately estimated. Each serum sample was divided into two aliquots: one diluted with phosphate-buffered saline and the other diluted with 1 M guanidine. We assayed the aliquots with the automated AxSYM HIV1/2gO test (Abbott Diagnostics Division), without modifying the manufacturer's protocol. We then calculated the avidity index (AI): the ratio of the sample/cutoff value for the guanidine aliquot to that of the phosphate-buffered saline aliquot. We analyzed 216 serum samples: 34 samples were collected within 6 months of seroconversion (recent seroconversions), and 182 were collected after 6 months. The mean AIs, by time from seroconversion, were 0.68 +/- 0.16 (within 6 months) and 0.98 +/- 0.10 (after 6 months) (P < 0.0001). AI of <0.90 correctly identified 88.2% of recent infections but misclassified as recent infections 13.2% of serum samples collected afterward. The probability of an infection being classified as recent and having AI of > or = 0.90 would be 0.7% in a population with 5% recent infections. AI can identify with a certain degree of accuracy recent HIV infections, and being a quantitative index, it provides different levels of sensitivity and specificity, depending on the selected cutoff value. The standard assay procedure is not modified. This test is simple and inexpensive and could be used for surveillance, decision-making in treatment, and prevention. 相似文献
95.
96.
Freer G Giannecchini S Tissot A Bachmann MF Rovero P Serres PF Bendinelli M 《Virology》2004,322(2):360-369
Immunogenicity of the tryptophan-rich motif (TrpM) in the membrane-proximal ectodomain of the transmembrane (TM) glycoprotein of feline immunodeficiency virus (FIV) was investigated. Peptide 59, a peptide containing the TrpM of the TM of FIV, was covalently coupled to Qbeta phage virus-like particles (Qbeta-59) in the attempt to induce potent anti-TrpM B cell responses in cats. All Qbeta-59 immunized cats, but not cats that received a mixture of uncoupled Qbeta and peptide 59, developed antibodies that reacted with a same epitope in extensive binding and binding competition assays. The epitope recognized was composed of three amino acids, two of which are adjacent. However, Qbeta-59-immune sera failed to recognize whole FIV in all binding and neutralization assays performed. Furthermore, no reactivity against the TrpM was detected by screening sera from FIV-infected cats that had reacted with TM peptides, confirming that this epitope does not seem to be serologically functional in the FIV virion. The data suggest that TrpM may not be a suitable target for antiviral vaccine design. 相似文献
97.
98.
Parra ER Canzian M Saber AM Coêlho RS Rodrigues FG Kairalla RA de Carvalho CR Capelozzi VL 《Pathology, research and practice》2004,200(10):701-705
Previous reports indicate that enlarged hilar and mediastinal lymph nodes caused by sarcoid-like reactions may develop after curative resection of cancer, and their presence does not necessarily denote neoplastic recurrence. Reports further suggest that coexisting pulmonary infiltrates in this setting may be related to sarcoidosis. In this study, we describe two patients who had resected lung and gastric cancer and who later developed pulmonary interstitial infiltrate, concurrent with progressive mediastinal lymphadenopathy initially thought to be caused by intrathoracic dissemination of their cancer. These changes were shown by open lung biopsy to be a benign, granulomatous reaction interpreted as sarcoidosis. Thus, it is important to recognize this clinical pattern when pulmonary infiltrates develop after complete treatment of cancer in an otherwise relapse-free patient and to encourage lung or lymph node biopsy in these particular settings in order to confirm a sarcoid-like reaction, thereby avoiding unnecessary chemotherapy for presumed tumor recurrence. 相似文献
99.
Gil LH Alves FP Zieler H Salcedo JM Durlacher RR Cunha RP Tada MS Camargo LM Camargo EP Pereira-da-Silva LH 《Journal of medical entomology》2003,40(5):636-641
Studies on seasonal anopheline fauna variation were performed in two distinct settlements in the State of Rond?nia, Brazil: one at the Madeira River banks (Portuchuelo) with stable native Amazonian population; the other at an inland lumber-extracting farm (Urupá) in dry land, in which adults are mostly migrants. During a 6-yr period (1994-2000), 8,638 adult anophelines were collected: 2,684 in Urupá and 5,954 in Portuchuelo. Anopheles darlingi represented >95% of total mosquitoes caught. Dissection of 4,424 A. darlingi females yielded a very low sporozoite infection index below 0.1%. Oocysts were found in both localities in approximately 0.1% of dissected mosquitoes. Determination of the hour biting rates disclosed seasonal variations in both localities. However, in Portuchuelo, mosquito density peaked at the acme of the rainy season, whereas at Urupá it peaked in the dry season. The increase in mosquito density and incidence of malaria cases were coincident. The high mosquito densities observed in the riverine settlement of Portochuelo sector B, which permits evaluation in > 10,000 mosquitoes' bites/person/year, could explain, in spite of the low mosquito's infection index, the previously described development of natural immunity in the local population that is not observed in the dry land agroindustrial settlement of Urupá. 相似文献
100.
Merati G Negrini S Sarchi P Mauro F Veicsteinas A 《European journal of applied physiology》2001,85(1-2):41-48
The use of a school backpack is one of the possible causes of back pain in children. Oxygen consumption (
), pulmonary ventilation, and heart rate (f
c) were measured in 35 pre-pubertal subjects [17 girls and 18 boys, mean (SD) age 11.3 (0.6) years]. They took part in a four-step
experiment: (1) standing for 5 min, (2) walking at 3 km·h–1 for 7 min, (3) walking at 3 km·h–1 for 7 min carrying a school backpack weighing 8 kg, and (4) walking at 7 km·h–1 for 5 min with no load. The occurrence of back pain in the last 2–3 years and during the last 15 days was assessed for the
subjects by means of a questionnaire. Mean (SD) standing
was 215 (45) ml.min–1 during walking at 3 km·h–1, 503 (101) ml.min–1 during walking without a load, and increased to 541 (98) ml.min–1 during walking with a load (P<0.01). Carrying a backpack increased f
c only minimally. The energy cost of walking at 3 km.h–1 without the backpack was 10.0 (2.0) ml O2
.m–1, and with the backpack was 10.8 (1.9) ml O2
.m–1 (P<0.01). The net energy cost of locomotion was 0.129 (0.032) ml.kg body mass–1.m–1 for the unloaded condition and slightly lower, at 0.123 (0.025) ml.kg body mass–1.m–1 during loaded walking (P<0.05). Ventilation did not change significantly between unloaded and loaded conditions. When the data were assessed according
to the occurrence of back pain, the f
c/
slope was significantly lower in children without back pain, even though the net energy cost of locomotion was similar. Overall,
these data suggest that the cardiovascular effortrequired for locomotion while carrying a backpack is minimal. However, fatigability
and back pain are more likely to take place in less physical performing subjects. Thus, the occurrence of back pain in schoolchildren
during locomotion while carrying a backpack may improve with an improvement in their level of fitness.
Electronic Publication 相似文献