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31.
The effects of antioxidant supplementation during Percoll preparation on human sperm DNA integrity 总被引:20,自引:6,他引:20
Hughes CM; Lewis SE; McKelvey-Martin VJ; Thompson W 《Human reproduction (Oxford, England)》1998,13(5):1240-1247
The integrity of sperm DNA is crucial for the maintenance of genetic
health. A major source of damage is reactive oxygen species (ROS)
generation; therefore, antioxidants may afford protection to sperm DNA. The
objectives of the study were, first, to measure the effects of antioxidant
supplementation in vitro on endogenous DNA damage in spermatozoa using the
single cell gel electrophoresis (comet) assay and, second, to assess the
effect of antioxidant supplementation given prior to X-ray irradiation on
induced DNA damage. Spermatozoa from 150 patients were prepared by Percoll
centrifugation in the presence of ascorbic acid (300, 600 microM), alpha
tocopherol (30, 60 microM), urate (200, 400 microM), or acetyl cysteine (5,
10 microM). DNA damage was induced by 30 Gy X-irradiation. DNA strand
breakage was measured using the comet assay. Sperm DNA was protected from
DNA damage by ascorbic acid (600 microM), alpha tocopherol (30 and 60
microM) and urate (400 microM). These antioxidants provided protection from
subsequent DNA damage by X-ray irradiation. In contrast, acetyl cysteine or
ascorbate and alpha tocopherol together induced further DNA damage.
Supplementation in vitro with the antioxidants ascorbate, urate and alpha
tocopherol separately has beneficial effects for sperm DNA integrity.
相似文献
32.
Development and application of genetic probes for detection of Enterocytozoon bieneusi in formalin-fixed stools and in intestinal biopsy specimens from infected patients. 总被引:2,自引:0,他引:2
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A Carville K Mansfield G Widmer A Lackner D Kotler P Wiest T Gumbo S Sarbah S Tzipori 《Clinical and Vaccine Immunology : CVI》1997,4(4):405-408
The microsporidium Enterocytozoon bieneusi is closely linked to wasting and diarrhea in a high proportion of individuals with AIDS. However, its relative contribution to disease is uncertain because diagnosis until recently depended on procedures involving endoscopy. A sensitive PCR technique which amplifies a fragment of the small-subunit rRNA gene of E. bieneusi from formalin-fixed stool samples was developed. Of 80 formalin-fixed stool samples collected from 74 Zimbabweans and 6 U.S. patients who were human immunodeficiency virus positive, 50% tested positive for E. bieneusi by PCR, whereas 24% tested positive for E. bieneusi by light microscopy of trichrome-stained fecal smears. In addition, we describe an in situ hybridization technique which detected and identified E. bieneusi as the causative agent in all six intestinal biopsy specimens tested. Both the PCR and in situ hybridization procedures are sensitive diagnostic tools which will complement currently available techniques and enable the differentiation of E. bieneusi from other microsporidia to be made. 相似文献
33.
Sheoran AS Feng X Singh I Chapman-Bonofiglio S Kitaka S Hanawalt J Nunnari J Mansfield K Tumwine JK Tzipori S 《Clinical and diagnostic laboratory immunology》2005,12(9):1109-1113
Enterocytozoon bieneusi is clinically the most significant among the microsporidia infecting humans, causing chronic diarrhea, wasting, and cholangitis in individuals with human immunodeficiency virus/AIDS. The lack of immune reagents is largely due to the absence of methods for laboratory propagation of E. bieneusi. We recently described a procedure for the concentration and purification of spores from diarrheic stool of infected humans. Purified spores were used to immunize mice for production and screening of monoclonal antibodies (MAbs) against E. bieneusi. The eight immunoglobulin M MAbs generated and fully characterized did not cross-react with other human microsporidia or with other microorganisms normally present in stool. One of the MAbs, 2G4, reacted with E. bieneusi spores in stools from monkeys and humans, without background fluorescence, which makes it an ideal diagnostic reagent. It also recognizes intracellular stages of the parasite and will be suitable for determining tissue distribution of E. bieneusi in infected hosts. At least two immunodominant antigens of E. bieneusi of 33,000 and 35,000 Da exist, which were recognized by rabbit and mouse antisera. The availability of MAbs against E. bieneusi will simplify considerably the diagnosis of this infection in humans and will provide tools for epidemiologic investigations regarding the true prevalence of the infection in various human and mammalian populations and the environmental sources of infection. 相似文献
34.
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
35.
Astrid LA. Kuijpers Rolph Pfundt Patrick LJM Zeeuwen Henri OF. Molhuizen Edwin CM. Mariman Peter CM. van de Kerkhof Joost Schalkwijk 《Clinical genetics》1998,54(1):96-101
Psoriasis is a multifactorial skin disease characterised by epidermal abnormalities and infiltration by lymphocytes and polymorphonuclear leukocytes (PMN). Skin-derived antileukoproteinase (SKALP), also known as elafin, is a potent inhibitor of human leukocyte elastase and proteinase 3, two PMN-derived proteinases implicated in tissue destruction and leukocyte migration. We have shown that, at least at the protein level, SKALP is significantly decreased in lesional skin of patients with pustular psoriasis compared with plaque-type psoriasis. This finding raised the possibility that SKALP could be one of the candidate genes for pustular forms of psoriasis. We therefore performed single strand conformation polymorphism (SSCP) analysis on the SKALP gene to screen for mutations/polymorphisms in the exons of 30 patients with plaque-type psoriasis, 15 patients with pustular psoriasis and 48 healthy controls. In exon 1 a polymorphism was detected at position + 43 relative to the translation start site, resulting in a substitution of threonine for alanine in the signal peptide. In the promoter region a dinucleotide repeat polymorphism was identified. Both polymorphisms were not associated with pustular psoriasis, or psoriasis in general. Our data indicate that the decrease in SKALP activity in pustular psoriasis is not caused by mutations in the coding region of the gene, and that there is no allelic association between pustular psoriasis and SKALP gene polymorphisms. 相似文献
36.
Association of DLG5 R30Q variant with inflammatory bowel disease 总被引:6,自引:0,他引:6
Daly MJ Pearce AV Farwell L Fisher SA Latiano A Prescott NJ Forbes A Mansfield J Sanderson J Langelier D Cohen A Bitton A Wild G Lewis CM Annese V Mathew CG Rioux JD 《European journal of human genetics : EJHG》2005,13(7):835-839
Crohn's disease (CD) and ulcerative colitis (UC) are chronic inflammatory diseases of the gastrointestinal system known as the inflammatory bowel diseases (IBD). Recently, Stoll and colleagues reported a novel finding of genetic variation in the DLG5 gene that is associated with IBD (CD and UC combined). We present here a study of the genetic variation described in that report in two well-powered, independent case-control cohorts and one family-based collection, and confirm the proposed association between IBD and the R30Q variant of DLG5 in two of the three studies. We are, however, unable to replicate the other proposed association to the common haplotype described in Stoll et al and suggest that this other finding could conceivably have been partially a statistical fluctuation and partially a result of LD with the replicated R30Q association. This study provides support for the hypothesis that DLG5 constitutes a true IBD risk factor of modest effect. 相似文献
37.
Five hundred allergy clinic patients were prick skin tested with papain, 1 mg/mL, in addition to usual local aeroallergens. Five of 475 subjects with seasonal allergic disease had positive skin tests to both papain and local pollens. None of the 25 individuals with negative skin test to pollens had skin reactivity to papain. The five subjects with positive skin tests to papain underwent double-blind placebo-papain challenges. All papain challenges were positive. Placebo challenges were negative. Papain-induced symptoms included palatal itching, watering itchy eyes, sneezing, rhinorrhea, abdominal cramps, diarrhea, and diaphoresis. Circulating papain-specific IgE was detected in all the papain-sensitive individuals, but not in control subjects. Confirmed papain sensitivity occurred in 1.05% of allergic subjects. In the papain-sensitive patients, cross-reacting antibodies with chymopapain were found. The small number of non-allergic subjects did not show any papain or chymopapain sensitivity in vitro. 相似文献
38.
Dal Zotto L; Quaderi NA; Elliott R; Lingerfelter PA; Carrel L; Valsecchi V; Montini E; Yen CH; Chapman V; Kalcheva I; Arrigo G; Zuffardi O; Thomas S; Willard HF; Ballabio A; Disteche CM; Rugarli EI 《Human molecular genetics》1998,7(3):489-499
We have recently reported isolation of the gene responsible for X- linked
Opitz G/BBB syndrome, a defect of midline development. MID1 is located on
the distal short arm of the human X chromosome (Xp22. 3) and encodes a
novel member of the B box family of zinc finger proteins. We have now
cloned the murine homolog of MID1 and performed preliminary expression
studies during development. Mid1 expression in undifferentiated cells in
the central nervous, gastrointestinal and urogenital systems suggests that
abnormal cell proliferation may underlie the defect in midline development
characteristic of Opitz syndrome. We have also found that Mid1 is located
within the mouse pseudoautosomal region (PAR) in Mus musculus , while it
seems to be X- specific in Mus spretus. Therefore, Mid1 is likely to be a
recent acquisition of the M. musculus PAR. Genetic and FISH analyses also
demonstrated a high frequency of unequal crossovers in the murine PAR,
creating spontaneous deletion/duplication events involving Mid1. These data
provide evidence for the first time that genetic instability of the PAR may
affect functionally important genes. In addition, we show that MID1 is the
first example of a gene subject to X-inactivation in man while escaping it
in mouse. These data contribute to a better understanding of the molecular
content and evolution of the rodent PAR.
相似文献
39.
40.
Harry S. Spaulding Jr. M.D.COL MC Lyndon E. Mansfield M.D.LTC MC Mark R. Stein M.D. John C. Sellner M.D. Daniel E. Gremillion M.D. 《The Journal of allergy and clinical immunology》1982,69(6):516-521
A double-blind modification of the intraesophageal acid perfusion challenge (Bernstein procedure) was performed in asthmatic subjects with and without gastroesophageal reflux, nonasthmatic subjects with reflux, and normal subjects. Conventional spirometric functions and total respiratory resistance (Rrs) were measured prior to and after the infusion. There were no changes in pulmonary functions except in the asthmatic subjects who had had a positive add challenge. The greatest changes occurred in Rrs, which increased significantly with reflux symptoms (p < 0.01) and decreased toward baseline (p < 0.05) when these symptoms were relieved with antacids. The response was even greater in asthmatic subjects who associated reflux symptoms with attacks of asthma. These results support previous findings that acid reflux symptoms could cause a bronchoconstrictive response in certain asthmatic patients. 相似文献