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41.
A conserved protein network controls assembly of the outer kinetochore and its ability to sustain tension 总被引:5,自引:0,他引:5
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Cheeseman IM Niessen S Anderson S Hyndman F Yates JR Oegema K Desai A 《Genes & development》2004,18(18):2255-2268
Kinetochores play an essential role in chromosome segregation by forming dynamic connections with spindle microtubules. Here, we identify a set of 10 copurifying kinetochore proteins from Caenorhabditis elegans, seven of which were previously uncharacterized. Using in vivo assays to monitor chromosome segregation, kinetochore assembly, and the mechanical stability of chromosome-microtubule attachments, we show that this copurifying protein network plays a central role at the kinetochore-microtubule interface. In addition, our analysis suggests that the network is comprised of three groups of proteins that contribute in distinct ways to this interface: KNL proteins act after the assembly of centromeric chromatin to generate the core of the microtubule-binding interface, MIS proteins control the rate and extent of formation of this interface, and NDC proteins are necessary to sustain tension during interactions with spindle microtubules. We also purify a similar set of associated proteins from human cells that includes four novel proteins and has recognizable homologs from each functional class. Thus, this protein network is a conserved constituent of the outer kinetochore, and the functions defined by our analysis in C. elegans are likely to be widely relevant. 相似文献
42.
Morphological evaluation of human embryos and derivation of an embryo quality scoring system specific for day 3 embryos: a preliminary study 总被引:16,自引:0,他引:16
Desai NN Goldstein J Rowland DY Goldfarb JM 《Human reproduction (Oxford, England)》2000,15(10):2190-2196
A scoring system specific for day 3 embryos has not been extensively explored. Most IVF laboratories continue to grade embryos solely on the basis of cell number and percentage fragmentation as was traditionally done for day 2 embryos. Additional morphological features, some unique to day 3 embryos, may be useful in selecting embryos most likely to blastulate and implant. The objective of this study was to derive an embryo scoring system for day 3 transfers which is predictive of positive pregnancy outcomes. A total of 316 transferred embryos from 93 patients was recorded on videotape and evaluated. The following parameters were used to grade the embryos: cell number, fragmentation pattern (FP), cytoplasmic pitting, compaction, equal sized blastomeres, blastomere expansion and absence of vacuoles. The clinical pregnancy rate was 41.9%, with an implantation rate of 18% per embryo transferred. The mean number of embryos transferred per patient was 3.4. Three formulae were derived to score embryo quality in each transfer based on the average score of individual embryos transferred. In the first scoring system, cell number alone was used to predict pregnancy outcome. The second scoring system was based on blastomere number and the observed FP. The third scoring system utilized both blastomere number and FP but also combined this with five morphological criteria to yield a final day 3 embryo quality (D3EQ) score. We found the D3EQ score to be prognostic of pregnancy outcome. This study suggests that although cell number and FP are certainly predictors of positive pregnancy outcomes, additional parameters specific to day 3 embryos should be used to stratify a cohort of embryos further. 相似文献
43.
134 swabs in viral transport medium were received from 126 patients with suspected clinical HSV-1 and HSV-2 infections. They were tested by (i) nested multiplex polymerase chain reaction NMPCR (strongly positive specimens had visible bands on both rounds of PCR) without prior extraction, (ii) culture in primary rhesus monkey kidney, E6-Vero, RD and HEp-2 cells and (iii) antigen detection by immunofluorescence (IF). Antigen detection employed four novel pools (A-D) of monoclonal antibodies (Mab): A was HSV-1 specific, B was HSV-2 specific while C and D were generic. In comparison to NMPCR the sensitivity and specificity of (i) culture was 59% (22/37) and 100% (134/134), (ii) IF by Pool A was 59% (16/27) and 100% (117/117), (iii) IF by Pool B was 40% (4/10) and 100% (130/130) and (iv) IF by Pools C and D were 60% (18/30) and 100% (96/96). Specimens positive by culture were more likely to be strongly positive by NMPCR (chi2 P = 0.004). Typing by each method concurred on all occasions. NMPCR was cost effective, easier to perform and was the most sensitive method for HSV detection. It should become the method of choice for HSV diagnosis. 相似文献
44.
Fluorescent amplified fragment length polymorphism genotyping of Campylobacter jejuni and Campylobacter coli strains and its relationship with host specificity, serotyping, and phage typing 总被引:4,自引:0,他引:4
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Fluorescent amplified fragment length polymorphism (FAFLP) analysis was applied to 276 Campylobacter jejuni strains and 87 Campylobacter coli strains isolated from humans, pigs, cattle, poultry, and retail meats to investigate whether certain FAFLP genotypes of C. jejuni and C. coli are associated with a particular host and to determine the degree of association between FAFLP-defined genotypes and heat-stable serotypes and/or phage types. Within C. coli, the poultry strains clustered separately from those of porcine origin. In contrast, no evidence of host specificity was detected among C. jejuni strains. While C. coli strains show host specificity by FAFLP genotyping, C. jejuni strains that are genotypically similar appear to colonize a range of hosts, rather than being host adapted. Some serotypes and/or phage types (C. jejuni serotype HS18, phage type PT6, and serophage type HS19/PT2 and C. coli HS66, PT2, and HS56/PT2) were the most homogeneous by FAFLP genotyping, while others were more heterogeneous (C. jejuni HS5 and PT39, and C. coli HS24 and PT44) and therefore poor indicators of genetic relatedness between strains. The lack of host specificity in C. jejuni suggests that tracing the source of infection during epidemiological investigations will continue to be difficult. The lack of congruence between some serotypes and/or phage types and FAFLP genotype underlines the need for phenotypic testing to be supplemented by genotyping. This study also demonstrates how, in general, FAFLP generates "anonymous" genetic markers for strain characterization and epidemiological investigation of Campylobacter in the food chain. 相似文献
45.
Correlation between macrophage activation and bactericidal function and Mycobacterium leprae antigen presentation in macrophages of leprosy patients and normal individuals. 总被引:2,自引:0,他引:2
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The killing of Mycobacterium leprae by resting and gamma interferon (IFN-gamma)-activated macrophages in normal subjects and leprosy patients was assessed. Resting macrophages from normal individuals demonstrated the ability to kill M. leprae. For macrophages from tuberculoid patients, killing of M. leprae was only achieved in the presence of IFN-gamma, suggesting that initial T-cell activation occurs prior to the killing of M. leprae. In contrast, though activation with IFN-gamma rendered the lepromatous macrophages microbicidal, it failed to induce lymphocyte proliferation, suggesting a defect at either the antigen-presenting cell or the lymphocyte level or both. The concept that T-cell anergy is primarily due to lack of lymphokine generation was ruled out by our results, since responsiveness was restored in only a small proportion of lepromatous patients after exogenous lymphokine addition. In conclusion, this study demonstrated that killing and antigen presentation are two independent events. It appears that the ability of the macrophages per se to kill M. leprae may be of greater importance than lymphocyte-mediated activation for protection against M. leprae infection. 相似文献
46.
Permanent impairment of embryo development by hydrosalpinges 总被引:6,自引:9,他引:6
Recent reports suggest a deleterious effect of hydrosalpinges on pregnancy
outcome for in-vitro fertilization (IVF) and improvement following surgical
treatment. We compared the effect of hydrosalpinx on pregnancy outcome in
286 patients having 348 IVF cycles and followed the development of
untransferred embryos for 7 days to determine if hydrosalpinges affect
oocyte quality or embryo development. The delivery rate per retrieval was
significantly lower for patients with hydrosalpinx, but was restored by
surgical treatment to that of patients without hydrosalpinx. However, the
implantation rate per embryo transferred and normal blastulation of
untransferred embryos, which were significantly decreased in patients with
hydrosalpinx, and growth arrest and degeneration of untransferred embryos,
which were significantly increased compared to patients without
hydrosalpinx, were not restored by surgical treatment of hydrosalpinges. We
conclude that surgical treatment of hydrosalpinges decreases early
pregnancy loss and improves pregnancy outcome, possibly by diminishing
reversible deleterious effects exerted on the endometrium. As we have seen
in our laboratory, hydrosalpinges may have a permanent negative influence
on ovarian function, follicular development and oocyte quality since
implantation of transferred embryos and normal blastulation of
untransferred embryos remain low, and in-vitro growth arrest and
degeneration remain high despite surgical treatment of hydrosalpinges.
相似文献
47.
A Kowal-Vern B A Birdsong G Dizikes R Radvany S N Desai H Schumacher 《Archives of pathology & laboratory medicine》1990,114(7):676-678
We present, to our knowledge, the first extensively studied case of lymphoid L2 blast crisis of chronic myelogenous leukemia with a hand mirror cell (HMC) variant. Special stains revealed the leukemic cells to be terminal deoxynucleotidyl transferase positive by immunofluorescence and cytochemically positive for alpha-naphthyl acetate esterase and acid phosphatase (diffuse granular). Immunophenotyping identified the major leukemic cell population as B-cells that expressed CD10+, CD19+, and HLA-DR+. It was not possible to separate the HMC and the non-HMC leukemic population by gating various cell populations, dual staining, cytochemistry, or by terminal deoxynucleotidyl transferase. Gene rearrangements were observed in both Ig heavy-chain alleles and one T-cell antigen receptor gamma-subunit allele. The rearrangements occupied all of the cells, indicating that the HMC and non-HMC were of a common clonal origin. The patient had a mosaic karyotype, with 90% of the cells having t(9;22), t(8;14), and t(9;15) translocations, an additional chromosome 8, and deleted chromosomes 9 and 15. Antibodies to simian sarcoma-associated virus and baboon endogenous virus were isolated in the patient's peripheral blood plasma. 相似文献
48.
Ghazzawi IM; Sarraf MG; Taher MR; Khalifa FA 《Human reproduction (Oxford, England)》1998,13(2):348-352
A prospective study was carried out to compare the fertilizing capability
and pregnancy outcome following intracytoplasmic sperm injection (ICSI)
using spermatozoa obtained from ejaculates, or surgically from epididymis
or seminiferous tubules. A total of 77 ICSI cycles (one per patient) was
included. In all, 28 patients had severe oligoasthenoteratozoospermia, 19
patients had obstructive azoospermia and 30 patients had non-obstructive
azoospermia. The main outcome measures were fertilization rate per injected
metaphase II oocyte and the clinical pregnancy rate per embryo transferred
back to the female recipients. In patients with severe
oligoasthenoteratozoospermia, the fertilization and pregnancy rates were 79
and 25 %. In patients with obstructive azoospermia, for whom epididymal
spermatozoa were used, these were 75 and 28%, and in the non-obstructive
group for which testicular spermatozoa were used for injection, they were
69 and 21% respectively. These rates were not significantly different in
the three groups (P = 0.85 and P = 0.14 respectively), suggesting that
spermatozoa from the ejaculates and epididymal or testicular biopsies are
able to fertilize equally by using ICSI. Live birth per embryo transfer was
significantly reduced in patients with non-obstructive azoospermia compared
to the other two groups. The high abortion rate (50%) in the group in which
testicular spermatozoa were used raises doubts about the developmental
competence of such embryos.
相似文献
49.
Ante mortem diagnosis of human rabies using saliva samples: comparison of real time and conventional RT-PCR techniques. 总被引:1,自引:0,他引:1
T Nagaraj Joel P Vasanth Anita Desai Anupa Kamat S N Madhusudana V Ravi 《Journal of clinical virology》2006,36(1):17-23
BACKGROUND: Rabies is an enzootic and fatal disease and is still a major problem in developing countries. Ante mortem diagnosis in human cases is necessary for medical management of the patient and to ensure appropriate post-exposure treatment of contacts. Both conventional RT-PCR and Real time PCR (TaqMan) have been described for the detection of rabies virus RNA from saliva and tissue respectively, however to date, there have been no studies comparing conventional and real time PCR assays for detection of rabies virus nucleic acid using saliva samples for ante mortem diagnosis. OBJECTIVES: In this study, we evaluated the utility of conventional RT-PCR and SYBR Green I Real time PCR in the ante mortem diagnosis of rabies using saliva samples. STUDY DESIGN: Saliva samples collected from twenty-four patients presenting with typical clinical manifestations of rabies were tested in the two assays. RESULTS: Amongst the 24 samples tested, 21 samples (87.5%) were positive by either of the two molecular methods. Of these 21, rabies virus RNA was detected in 6/21 in the conventional RT-PCR assay while SYBR Green I Real time PCR could detect RNA in 18/21 samples. CONCLUSION: Real time PCR assay was more sensitive than conventional RT-PCR assay (sensitivity 75% versus 37%, p=0.0189). This study highlights the utility of molecular diagnostic tests in establishing ante mortem diagnosis of rabies using saliva samples within a few hours. 相似文献
50.
Jhingan HP Jain R Desai NG Vaswani M Tripathi BM Pandey RM 《Indian journal of medical sciences》2002,56(10):495-500
Self-report validity of recent drug use among heroin abusers depends on many factors including the population being studied and the setting in which the study is carried out. This study was conducted by the treating physicians to assess the self-report validity of recent heroin use by heroin dependent patients in the outdoor setting using 'thin layer chromatography' (TLC) and two highly sensitive methods of urinalysis viz. 'gas liquid chromatography' (GLC) and 'high performance liquid chromatography' (HPLC). Out of seventy-six heroin dependent patients who entered the study, 64 provided urine sample on the same day. Patients' self-report about recent opiate use was found to have a moderate agreement with urinalysis report. However, it is important to validate it with urinalysis during the treatment process because a substantial proportion of patients fails to report recent opiate use. It is recommended that all drug dependence treatment centres should be equipped with a sensitive urinalysis facility. Otherwise, the outcome of the treatment process should be considered with caution. 相似文献