Homologous and illegitimate recombination in developing Xenopus oocytes and eggs.

Exogenous DNA is efficiently recombined when injected into the nuclei of Xenopus laevis oocytes. This reaction proceeds by a homologous resection-annealing mechanism which depends on the activity of a 5'-->3' exonuclease. Two possible functions for this recombination activity have been proposed: it may be a remnant of an early process in oogenesis, such as meiotic recombination or amplification of genes coding for rRNA, or it may reflect materials stored for embryogenesis. To test these hypotheses, recombination capabilities were examined with oocytes at various developmental stages. Late-stage oocytes performed only homologous recombination, whereas the smallest oocytes ligated the restriction ends of the injected DNA but supported no homologous recombination. This transition from ligation to recombination activity was also seen in nuclear extracts from these same stages. Exonuclease activity was measured in the nuclear extracts and found to be low in early stages and then to increase in parallel with recombination capacity in later stages. The accumulation of exonuclease and recombination activities during oogenesis suggests that they are stored for embryogenesis and are not present for oocyte-specific functions. Eggs were also tested and found to catalyze homologous recombination, ligation, and illegitimate recombination. Retention of homologous recombination in eggs is consistent with an embryonic function for the resection-annealing mechanism. The observation of all three reactions in eggs suggests that multiple pathways are available for the repair of double-strand breaks during the extremely rapid cleavage stages after fertilization.     

Divergence in male mating tactics between two populations of the soapberry bug: I. Guarding versus nonguarding

I compared male allocation to prolonged mate guarding versusnot guarding between two populations of the soapberry bug (Jaderahaematoloma) that differ in adult sex ratio: Oklahoma, USA (mean± SD adult sex ratio, 2.70 ± 0.95 males per female),and Florida, USA (1.09 ± 0.26 males per female). To predictthe reproductive performance of each mating tactic in each population,I collected data on search time per mating, time required forguarding to be effective, sperm competition, female rematingpropensity, and female resistance to guarding. Search time alonediffered significantly between the populations, being much greaterin Oklahoma (estimated as 26.2 h per mate) than in Florida (estimatedas 9.6 h per mate). For males in each region, these data wereused to model the costs and benefits of guarding for differentnumbers of oviposition bouts versus not guarding. The reproductiverate of nonguarders in Oklahoma is exceeded by that of guarderswho remain with a female for more than one oviposition bout,but in Florida, the reproductive rate of nonguarders is onlyexceeded by that of guarders who remain with a female for atleast three ovipositions. Consistent with the model, Oklahomamales in field arenas guarded more frequently than did Floridamales. However, nonguarding was common in both populations,and guarding durations were highly variable.     

Serotype F double- and triple-converting phage insertionally inactivate the Staphylococcus aureus β-toxin determinant by a common molecular mechanism

Abstract The precise molecular mechanism of Staphylococcus aureus β -toxin inactivation by the serotype F triple-converting phage φ42, φA1 and φA3 was investigated. Sequence analysis of the φ42 ( attP ) and Staphylococcus aureus ( attB ) attachment sites and the left ( attL ) and right ( attR ) chromosomal/bacteriophage DNA junctions of individual lysogens, each harbouring a triple-converting phage, revealed the presence of a common 14-bp core sequence in all four sites. These findings indicate that the genomes of the triple-converting phage integrate into the 5'-end of the β-toxin gene ( hlb ) by a site- and orientation-specific mechanism identical to that previously described for the serotype F double-converting phage φ13.     

Use of the maize transposonsActivator andDissociation to show that phosphinothricin and spectinomycin resistance genes act non-cell-autonomously in tobacco and tomato seedlings

Cell-autonomous genes have been used to monitor the excision of both endogenous transposons in maize andAntirrhinum, and transposons introduced into transgenic plants. In tobacco andArabidopsis, the streptomycin phosphotransferase (SPT) gene reveals somatic excision of the maize transposonActivator (Ac) as green sectors on a white background in cotyledons of seedlings germinated in the presence of streptomycin. Cotyledons of tomato seedlings germinated on streptomycin-containing medium do not bleach, suggesting that a different assay for transposon excision in tomato is desirable. We have tested the use of the spectinomycin resistance (SPEC) gene (aadA) and a Basta resistance (BAR) gene (phosphinothricin acetyltransferase, or PAT) for monitoring somatic excision ofAc in tobacco and tomato. Both genetic and molecular studies demonstrate that genotypically variegated individuals that carry clones of cells from whichAc orDs have excised from either SPEC or BAR genes, can be phenotypically completely resistant to the corresponding antibiotic. This demonstrates that these genes act non-cell-autonomously, in contrast to the SPT gene in tobacco. Possible reasons for this difference are discussed.     

Inherited prion disease (PrP lysine 200) in Britain: two case reports.

OBJECTIVE--To identify cases of inherited prion diseases in Britain and to assess their phenotypic features. DESIGN--Screening study of patients suspected clinically to have Creutzfeldt-Jakob disease and other neurodegenerative diseases by prion protein gene analysis. SETTING--Biochemical research department. SUBJECTS--Patients suspected to have Creutzfeldt-Jakob disease and other neurodegenerative diseases. RESULTS--Two patients with symptoms characteristic of sporadic Creutzfeldt-Jakob disease were found to have inherited prion protein disease (PrP lysine 200), with a mutation at codon 200 of the prion protein gene. Both were homozygous at codon 129 of the gene. One patient was a man aged 58 of British descent while the other was of Libyan Jewish origin. CONCLUSION--Two foci of inherited prion disease are known, among Libyan Jews and in Slovakia. A separate British focus of the disease may also exist. Heterozygosity at codon 129 may lead to reduced penetrance of the mutation.     

Internal initiation of translation of hepatitis C virus RNA: the ribosome entry site is at the authentic initiation codon.

Hepatitis C viral (HCV) RNA includes an internal ribosome entry segment (IRES) that extends some 30 nt into the coding region and promotes internal initiation of translation at the authentic initiation codon at nt 342. The 5'-boundary of this IRES was mapped by in vitro translation and transfection assays and was found to lie between nt 42 and 71. Within these IRES boundaries there are, in most HCV strains, three AUG triplets upstream of the authentic initiation site. Although the first, 5'-proximal, of these is absolutely conserved, a mutational analysis showed that it is not a functional initiation codon. In particular, the G residue could be substituted provided compensatory mutations were made to maintain base pairing. The other two upstream AUGs are not absolutely conserved, and mutation of the third (5'-distal) had little effect on IRES activity. When an additional AUG codon was introduced by single-site mutation just upstream of the authentic initiation codon, it was found to be used when most of the IRES had been deleted to generate an RNA translated by the scanning ribosome mechanism, but was not used in the background of the full-length IRES when internal initiation is operative. These results argue that the IRES promotes direct ribosome entry immediately at, or indeed very close to, the authentic initiation codon, and that the upstream AUGs do not serve as ribosome entry sites.     

Female family physicians in obstetrics: achieving personal balance.

OBJECTIVE: To describe the experiences of female family physicians who practise obstetrics in balancing professional obligations with personal and family needs, given the unique challenges that such practice poses for these physicians. DESIGN: Qualitative study. SETTING: Ontario. PARTICIPANTS: A purposefully selected sample of nine female family physicians who met the criteria of being married, having children and currently practising obstetrics. OUTCOME MEASURES: Experiences of female family physicians and their strategies in their personal, family and professional lives that enable them to continue practising obstetrics. RESULTS: All participants continued to practise obstetrics because of the pleasure they derived from it, despite the challenges of balancing the unpredictable demands of obstetrics with their personal and family needs. To continue in obstetrics, they needed to make changes in their lives, either through a gradual, evolutionary process or in response to a critical event. Alterations to work and family arrangements permitted them to meet the challenges and led to increased satisfaction. Changes included making supportive call-group arrangements, limiting work hours and the number of births attended and securing help with household duties. CONCLUSIONS: An in-depth examination, through the use of qualitative methods, showed the reasons why some female family physicians continue to practise obstetrics despite the stressful aspects of doing so. This knowledge may be useful for women who are residents or experienced clinicians and who are considering including obstetrics in their practice.