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51.
Rapid detection of methicillin-resistant Staphylococcus aureus directly from sterile or nonsterile clinical samples by a new molecular assay 总被引:13,自引:0,他引:13
Francois P Pittet D Bento M Pepey B Vaudaux P Lew D Schrenzel J 《Journal of clinical microbiology》2003,41(1):254-260
A rapid procedure was developed for detection and identification of methicillin-resistant Staphylococcus aureus (MRSA) directly from sterile sites or mixed flora samples (e.g., nose or inguinal swabs). After a rapid conditioning of samples, the method consists of two main steps: (i) immunomagnetic enrichment in S. aureus and (ii) amplification-detection profile on DNA extracts using multiplex quantitative PCR (5'-exonuclease qPCR, TaqMan). The triplex qPCR assay measures simultaneously the following targets: (i) mecA gene, conferring methicillin resistance, common to both S. aureus and Staphylococcus epidermidis; (ii) femA gene from S. aureus; and (iii) femA gene from S. epidermidis. This quantitative approach allows discrimination of the origin of the measured mecA signal. qPCR data were calibrated using two reference strains (MRSA and methicillin-resistant S. epidermidis) processed in parallel to clinical samples. This 96-well format assay allowed analysis of 30 swab samples per run and detection of the presence of MRSA with exquisite sensitivity compared to optimal culture-based techniques. The complete protocol may provide results in less than 6 h (while standard procedure needs 2 to 3 days), thus allowing prompt and cost-effective implementation of contact precautions. 相似文献
52.
The effects of co-culture with human fibroblasts on human embryo development in vitro and implantation 总被引:5,自引:0,他引:5
Wetzels AM; Bastiaans BA; Hendriks JC; Goverde HJ; Punt-van der Zalm AP; Verbeet JG; Braat DD 《Human reproduction (Oxford, England)》1998,13(5):1325-1330
In a human in-vitro fertilization (IVF) programme, the effect of co-
culture of embryos with human fibroblasts was evaluated with respect to
pregnancy rate and embryo development. Patients were included in the study
after giving informed written consent. The IVF treatments were randomly
assigned by stratification of both age (<36 versus > or =36 years)
and previous IVF attempts (yes versus no). After fertilization was
established, the zygotes were transferred to a 4-well dish with or without
fibroblasts and cultured for 2 days. On the third day after ovum pick-up
(OPU), cell number and quality [5 (good) to 1 (poor)] of the embryos were
scored and a maximum of three embryos was transferred. Supernumerary
embryos of good quality were cryopreserved. The design of this study was a
group sequential trial with the objective of detecting differences between
pregnancy rates following IVF with conventional incubation or incubation in
co-culture with fibroblasts. This design included one evaluation at
half-way data collection. In the study, 148 patients had an OPU, of whom 77
were allocated to the co-culture group. There was no statistically
significant difference in pregnancy rate, cell number and embryo quality
between the two groups. The ongoing pregnancy rate per embryo transfer was
27% in co-culture and 30% in the conventional culture group. The
implantation rates per transferred embryo were 17 and 18% respectively.
Using a multivariate logistic regression model for the probability of
ongoing pregnancies, the odds ratio of co-culture, adjusted for age and
previous IVF attempts, was not statistically significant. In conclusion,
co-culture with human fibroblasts does not contribute to an improvement of
embryo quality nor to a higher pregnancy rate after IVF in an unselected
group of patients.
相似文献
53.
Expression and self-assembly of recombinant capsid protein from the antigenically distinct Hawaii human calicivirus. 总被引:4,自引:7,他引:4 下载免费PDF全文
K Y Green A Z Kapikian J Valdesuso S Sosnovtsev J J Treanor J F Lew 《Journal of clinical microbiology》1997,35(7):1909-1914
The Norwalk and Hawaii viruses are antigenically distinct members of the family Caliciviridae and are considered to be important etiologic agents of epidemic gastroenteritis, with most studies focusing on the role of Norwalk virus. To further investigate the importance of Hawaii virus, Hawaii virus-like particles (VLPs) were produced by expression of its capsid protein in the baculovirus system and these VLPs were used as the antigen in an enzyme-linked immunosorbent assay that was efficient in the detection of a serologic response to Hawaii virus. The ready availability of Hawaii VLPs should enable larger-scale epidemiological studies to further elucidate the importance of this agent. 相似文献
54.
55.
56.
Anderson RA; Wallace AM; Kicman AT; Wu FC 《Human reproduction (Oxford, England)》1997,12(8):1657-1662
Administration of supraphysiological doses of testosterone to normal men
causes inhibition of spermatogenesis, but while most become azoospermic,
30-55% maintain a low rate of spermatogenesis. We have investigated whether
there are differences in endogenous androgen production, of testicular and
adrenal origin, which may be related to the degree of suppression of
spermatogenesis. Thirty-three healthy Caucasian men were given weekly i.m.
injections of 200 mg testosterone oenanthate (TE), 18 became azoospermic,
while 15 remained oligozoospermic. Urinary excretion of epitestosterone, a
specific testicular product, was reduced to <10% of pretreatment values,
with no differences between the groups. Similar results were obtained for
other markers of testicular steroidogenesis. Urinary and plasma adrenal
androgens were also reduced during TE treatment: a statistically
significant decrease in both (P < 0.001 and P < 0.05 respectively)
was seen in the azoospermic but not oligozoospermic responders. These
results suggest that testicular steroidogenesis is decreased to <10% by
the administration of supraphysiological doses of exogenous testosterone.
Differences in the degree of ongoing steroidogenesis in the testis do not
appear to account for incomplete suppression of spermatogenesis, thus
differences in androgen metabolism may underlie this heterogeneous
response. A small but significant reduction in secretion of adrenal
androgens was also detectable, the relevance of which is unclear.
相似文献
57.
Zhan Y Brown LE Deliyannis G Seah S Wijburg OL Price J Strugnell RA O'Connell PJ Lew AM 《Immunologic research》2004,30(1):1-14
The most common models of CD4 T-cell deficiency are mice exogenously injected with anti-CD4 antibody (Ab), CD4 knockout (CD4−/−) and major histocompatibility complex (MHC) class II knockout (class II−/−) mice. We recently described the anti-CD4 Ab transgenic mouse (GK) as an improved CD4 cell-deficient model. This review compares
this new GK mouse model with the widely available class II−/− and CD4−/− mice, when exposed to complex antigens (foreign grafts and during bacterial or viral infection). We highlight here the cytometric
and functional differences (including Ab isotype, viral or bacterial clearance, and graft survival) among these CD4 cell-deficient
models. For example, whereas grafts are generally rejected in class II−/− and CD4−/− mice as quickly as in wild-type mice, they survive longer in GK mice. Also, CD4−/− mice produce IgG against both simple model and complex antigens, but class II−/− and GK mice produce small amounts of IgG2a against complex antigens but not simple model antigens. These differences harbinger
the caveats in the use of these various mice. 相似文献
58.
N E Morton D C Rao H Lang-Brown C J Maclean R D Bart R Lew 《Journal of medical genetics》1977,14(1):1-9
This reanalysis of a classic survey leads to inferences about design of genetic studies, resolution of heterogeneity, and the role of autosomal and sex-linked genes in mental retardation, which is no longer refractory to segregation analysis. By discriminating between sociofamilial and biological types we estimate that at least 351 autosomal loci can produce mental retardation, with an inbred load of 0.83 detrimental equivalents and a mutation rate of 0.008 per gamete, or less than 2.4 X 10(-5) per locus. The distribution of probands was estimated as: 7 per cent medical, 60 per cent sociofamilial, and 33 per cent biological. Simple genetic mechanisms account for virtually all the biological category. Within the sociofamilial group cultural inheritance and polygenes could not be resolved. 相似文献
59.
Time course of hypo-osmotic swellings of human spermatozoa: evidence of ordered transition between swelling subtypes 总被引:2,自引:1,他引:2
Hossain AM; Rizk B; Barik S; Huff C; Thorneycroft IH 《Human reproduction (Oxford, England)》1998,13(6):1578-1583
The hypo-osmotic swelling test (HOST or HOS test) usually takes into
consideration the total HOS response value with no emphasis either on the
value of the response subtypes or the response evaluation time. This study
investigated the time course of HOS responses and analysed their
physiological relevance. Raw semen spermatozoa and Percoll washed
spermatozoa were used in the experiment. The morphological changes in the
sperm tail were monitored by incubating the spermatozoa in the hypo-
osmotic solution for 16 different time periods. The HOS reactive
spermatozoa and the type of HOS reaction (swelling subtypes) of the samples
subjected to different duration of treatment were identified under a phase
contrast microscope. Also the fate of individual spermatozoa in a
hypo-osmotic environment were monitored for 30 min. In spermatozoa exposed
to a hypo-osmotic solution, the motility lasted usually less than 2 min and
motility characteristics were uniquely different from that of the
spermatozoa under iso-osmotic conditions. The HOS response development was
permanent but the motility loss due to hypo-osmotic shock was reversible up
to 1 min of incubation. There was an indication of ordered transition among
the HOS swelling subtypes apparently initiating with subtype b destined to
c, d, e, f and g. Further, the subtypes a and g showed gradual decrease and
increase, respectively, while subtype b showed abrupt initial increase and
then gradual decrease. Transition from b to g could be direct or via one or
more than one subtypes. Ultrastructure based analysis indicated that HOS
response subtypes are the apparent reflection of the differences in the
cytoskeletal assembly of the sperm tail and thus may be identifying
different physiological variants in the sperm population. These results
indicate that shorter incubation is essential to document the kinetics of
various HOS responses but the conventional HOS test misses these important
HOS features because of lengthy incubation. Since the time course of
ordered transition of HOS responses will vary more than the total HOS
response in semen of different aetiologies, the importance of HOS response
subtypes and response evaluation time should be taken into consideration
when applying HOS test.
相似文献
60.
The effect of ovarian steroids on epithelial ciliary beat frequency in the human Fallopian tube 总被引:3,自引:3,他引:3
Mahmood T; Saridogan E; Smutna S; Habib AM; Djahanbakhch O 《Human reproduction (Oxford, England)》1998,13(11):2991-2994
Using a method that detects variations in light intensity we have studied
the effect of ovarian steroids on human Fallopian tube epithelial ciliary
beat frequency in vitro. We have found that baseline ciliary beat frequency
averages between 5-6 Hz. Cilia from ampullary segments of the Fallopian
tube beat significantly faster (5.4 Hz+/-0.2) than those from fimbrial
segments (4.8 Hz+/-0.2). There was no significant difference in baseline
ciliary beat frequency at any other anatomical site in the Fallopian tube.
Incubation with progesterone (10 micromol/l) suppresses human Fallopian
tube epithelial ciliary beat frequency by 40-50%. This inhibition was
observed at similar magnitudes in all Fallopian tubes studied irrespective
of anatomical site. Progesterone-induced reductions in ciliary beat
frequency were concentration dependent and prevented by the progesterone
receptor antagonist mifepristone (RU486). Oestradiol alone (10 micromol/l)
had no effect on ciliary beat frequency at any anatomical site in the
Fallopian tube but did prevent the reduction in ciliary beat frequency seen
with progesterone when tissues were incubated with these two steroids
together.
相似文献