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Evaluation of three substitutes for Percoll in sperm isolation by density gradient centrifugation 总被引:9,自引:4,他引:9
Silane-coated silica particle solutions (ISolate(TM) and PureSperm)TM)) and
iodixanol (OptiPrep(TM)) were compared to polyvinylpyrrolidone (PVP)-coated
silica particles (Percoll(TM)) in their efficacy to recover spermatozoa by
gradient centrifugation for use in assisted reproductive procedures.
Efficacy was assessed in terms of percentages of sperm recovery, sperm
vitality and motility, normal sperm morphology and normal sperm chromatin
condensation. No significant difference was found in the recovery of
spermatozoa for men with both normal sperm counts and oligozoospermia,
between PVP-coated and silane-coated particle solutions. Iodixanol had
significantly lower sperm recovery compared to the other products. Sperm
vitality, progressive motility, normal morphology and normal chromatin
condensation did not differ significantly between any of the sperm
isolation products.
相似文献
6.
An extracorporeal circulation technique was developed for use in rats to provide equilibrated blood samples for multiple hormone assays. The inclusion of the extracorporeal circulation did not significantly alter arterial blood pressure, cardiac output, heart rate or central venous pressure in either Brattleboro rats with hereditary diabetes insipidus (BDI) or normal rats of the parent Long Evans (LE) strain. Plasma adrenaline and noradrenaline levels did not alter in either BDI or LE rats following inclusion of the extracorporeal circulation but the vasopressin concentration rose significantly in the LE rats. The impaired recovery of the mean arterial blood pressure following haemorrhage in the BDI rats compared with normal LE animals was not further influenced by the inclusion of the extracorporeal circulation. Plasma vasopressin and adrenaline (but not nor-adrenaline) levels were significantly raised during, and after, haemorrhage in the LE rats while in the BDI rats only plasma adrenaline levels were significantly increased. These results show that the insertion of an extracorporeal circulation into an anaesthetized BDI or LE rat does not adversely affect the cardiovascular system despite the increase in baseline plasma vasopressin concentration in normal rats, and its subsequent removal provides an additional equilibrated blood sample for multiple hormone assay within the same animal. The increased release of both adrenaline and vasopressin (but not noradrenaline) after haemorrhage in the same animal is detected using this technique, and the importance of vasopressin to the normal recovery process confirmed. 相似文献
7.
Mutation spectrum and genotype-phenotype analyses in Cowden disease and Bannayan-Zonana syndrome, two hamartoma syndromes with germline PTEN mutation 总被引:22,自引:1,他引:22
Marsh DJ; Coulon V; Lunetta KL; Rocca-Serra P; Dahia PL; Zheng Z; Liaw D; Caron S; Duboue B; Lin AY; Richardson AL; Bonnetblanc JM; Bressieux JM; Cabarrot-Moreau A; Chompret A; Demange L; Eeles RA; Yahanda AM; Fearon ER; Fricker JP; Gorlin RJ; Hodgson SV; Huson S; Lacombe D; Eng C 《Human molecular genetics》1998,7(3):507-515
The tumour suppressor gene PTEN , which maps to 10q23.3 and encodes a 403
amino acid dual specificity phosphatase (protein tyrosine phosphatase;
PTPase), was shown recently to play a broad role in human malignancy.
Somatic PTEN deletions and mutations were observed in sporadic breast,
brain, prostate and kidney cancer cell lines and in several primary tumours
such as endometrial carcinomas, malignant melanoma and thyroid tumours. In
addition, PTEN was identified as the susceptibility gene for two hamartoma
syndromes: Cowden disease (CD; MIM 158350) and Bannayan-Zonana (BZS) or
Ruvalcaba-Riley-Smith syndrome (MIM 153480). Constitutive DNA from 37 CD
families and seven BZS families was screened for germline PTEN mutations.
PTEN mutations were identified in 30 of 37 (81%) CD families, including
missense and nonsense point mutations, deletions, insertions, a
deletion/insertion and splice site mutations. These mutations were
scattered over the entire length of PTEN , with the exception of the first,
fourth and last exons. A 'hot spot' for PTEN mutation in CD was identified
in exon 5 that contains the PTPase core motif, with 13 of 30 (43%) CD
mutations identified in this exon. Seven of 30 (23%) were within the core
motif, the majority (five of seven) of which were missense mutations,
possibly pointing to the functional significance of this region. Germline
PTEN mutations were identified in four of seven (57%) BZS families studied.
Interestingly, none of these mutations was observed in the PTPase core
motif. It is also worthy of note that a single nonsense point mutation,
R233X, was observed in the germline DNA from two unrelated CD families and
one BZS family. Genotype-phenotype studies were not performed on this small
group of BZS families. However, genotype-phenotype analysis inthe group of
CD families revealed two possible associations worthy of follow-up in
independent analyses. The first was an association noted in the group of CD
families with breast disease. A correlation was observed between the
presence/absence of a PTEN mutation and the type of breast involvement
(unaffected versus benign versus malignant). Specifically and more
directly, an association was also observed between the presence of a PTEN
mutation and malignant breast disease. Secondly, there appeared to be an
interdependent association between mutations upstream and within the PTPase
core motif, the core motif containing the majority of missense mutations,
and the involvement of all major organ systems (central nervous system,
thyroid, breast, skin and gastrointestinal tract). However, these
observations would need to be confirmed by studying a larger number of CD
families.
相似文献
8.
Lambert I.B.; Carroll C.; Laycock N.; Duval L.; Whiteway J.; Lawford I.; Turner G.; Booth R.; Douville S.; Nokhbeh M.R. 《Mutagenesis》1998,13(1):9-18
We have examined the mutational specificity of 1-nitroso-6-nitropyrene(1,6-NONP), an activated metabolite of the carcinogen 1,6-dinitropyrene,in the lacI gene of Escherichia coli strains which are deficientin nucleotide excision repair (strain NR6113, 相似文献
9.
The injection of antigen into the peritoneal cavities of actively sensitised rats produced an immediate reaction characterised by an increase in concentrations in the peritoneal fluids, collected 5 min later, of extravasated dye labelled plasma proteins, histamine and slow reacting substance of anaphylaxis (SRS-A). Changes were also produced in the numbers of leucocytes in the blood and peritoneal cavity. 5 min after antigen challenge there was a reduction in the number of cells that could be washed from the peritoneal cavity. 4 h after antigen there was an increase in numbers of neutrophils both in the blood and peritoneal washings and these fell to the levels in control rats at 24 h. 24 h after antigen, and continuing for 72 h, there was an increase in numbers of eosinophils and mononuclear cells in the peritoneal washings.The rats were injected intravenously with sephadex particles to produce a blood eosinophilia at the time of antigen challenge, this increased the numbers of eosinophils migrating into the peritoneal cavity but had no effect on antibody levels, the numbers of other leucocytes or on the immediate reaction.An inhibitor of lipoxygenase and cyclo-oxygenase metabolism of arachidonic acid, phenidone, at 100 mg/kg p.o., inhibited SRS-A release to control levels, in the immediate reaction, but had not effect on the leucocyte changes. The glucocorticosteroid, dexamethasone, at doses of 0.1 and 1 mg/kg p.o., produced little inhibition of SRS-A release but significantly inhibited neutrophil, eosinophil and mononuclear cell infiltration into the peritoneal cavity. These results suggest that arachidonic acid metabolites released in the immediate reaction are not the prime mediators of the cellular changes. Isoprenaline, at 0.05 and 0.2 mg/kg s.c., inhibited extravasation in the immediate reaction with no effect on histamine release but only the higher dose inhibited neutrophil and eosinophil infiltration into the peritoneal cavity. Aminophylline, at 50 mg/kg p.o., had no effect on the immediate reaction but inhibited the neutrophil infiltration. Disodium cromoglycate (DSCG) at 20 and 100 mg/kg s.c. inhibited the immediate reaction but this had no effect upon the cellular changes taking place after 5 min. Cyproheptadine at 1 mg/kg s.c. inhibited extravasation but had no effect on the cellular changes. It appears therefore that factors other than those derived from the mast cell were responsible for the cellular changes in this system. DSCG at 100 mg/kg s.c. and aminophylline at 25 and 50 mg/kg p.o. prevented the reduction in the number of cells that could be washed from the peritoneal acvity 5 min after antigen challenge. 相似文献
10.
We report our clinical experience with phototherapy in 3802 infants; 3629 were exposed to "standard" daylight phototherapy and 173 to "high-intensity" blue-light phototherapy. High-intensity blue-light phototherapy was twice as effective as standard daylight phototherapy in decreasing bilirubin concentrations. No failures occurred with high-intensity phototherapy compared with an overall failure rate of 1.84/1000 with daylight lamps; these cases were transferred to high-intensity phototherapy with prompt response. Rebound after cessation of phototherapy was greater in those exposed to high-intensity blue light with a significantly greater number requiring a second exposure. However, the incidence was still low. No third exposure was required in any infant. Nursing of infants under high-intensity blue light was more difficult and inconvenient as was clinical monitoring. The light also caused more stress on the nursing and medical personnel. However, the infants tolerated both types of phototherapy equally well. High-intensity blue-light phototherapy would seem to be the treatment of choice for infants with rapidly increasing or very high bilirubin levels, as well as in those not responding adequately to daylight phototherapy. 相似文献