Female genital cosmetic surgery: a cross-sectional survey exploring knowledge, attitude and practice of general practitioners.

OBJECTIVE: To explore general practitioner's (GP) knowledge, attitudes and practice regarding female genital cosmetic surgery (FGCS) in Australia. DESIGN: Cross-sectional survey. SETTING: Australia. SAMPLE: GPs who attended a women's health seminar and GPs who subscribed to a non-governmental, national health professional organisation database that provides education to primary care professionals. METHOD: A national online survey of GPs was conducted for the 10-week period, starting 1 week prior and 2 months after a Women's Health seminar was held in Perth on 8 August 2015. 31 questions prompted GPs' knowledge, attitudes and practice in managing patients asking about FGCS. RESULTS: The survey was fully completed by 443 GPs; 54% had seen patients requesting FGCS. Overall, 75% (95% CI 71% to 79%) of GPs rated their knowledge of FGCS as inadequate and 97% (95% CI 94% to 99%) had been asked by women of all ages about genital normality. Of those who had seen patients requesting FGCS, nearly half (44%, 95% CI 38% to 51%) reported they had insufficient knowledge of risks of FGCS procedures and 35% (95% CI 29% to 41%) reported seeing females younger than 18 years of age requesting FGCS. Just over half (56%, 95% CI 51% to 60%) of the GPs felt that women should be counselled before making a referral for FGCS. More than half the GPs suspected psychological disturbances in their patients requesting FGCS such as depression, anxiety, relationship difficulties and body dysmorphic disorder. CONCLUSIONS: GPs see women of all ages presenting with genital anatomy concerns and in those who request FGCS, GPs often suspected a range of mental health difficulties. GPs require greater education to support their patients who request FGCS.

Oncogenic pathways impinging on the G2-restriction point.

In the absence of mitogenic stimuli, cells normally arrest in G(1/0), because they fail to pass the G1-restriction point. However, abrogation of the G1-restriction point (by loss of the retinoblastoma gene family) reveals a second-restriction point that arrests cells in G2. Serum-starvation-induced G2 arrest is effectuated through inhibitory interactions of p27(KIP1) and p21(CIP1) with cyclins A and B1 and can be reversed through mitogen re-addition. In this study, we have investigated the pathways that allow cell cycle re-entry from this G2 arrest. We provide evidence that recovery from G2 arrest depends on the rat sarcoma viral oncogene (RAS) and phosphatidylinositol-3 kinase pathways and show that oncogenic hits, such as overexpression of c-MYC or mutational activation of RAS can abrogate the G2-restriction point. Together, our results provide new mechanistic insight into multistep carcinogenesis.

The metabolism of linoleic acid in healthy subjects after intake of a single dose of (13)C-linoleic acid.

OBJECTIVE: To investigate the in vivo oxidation of (13)C18:2n-6 and its conversion into longer-chain polyunsaturates (LCPs) in healthy subjects. DESIGN: Blood samples were collected from six subjects before (fasted) and 7, 11 (non-fasted), 24, 48, 72, 168 and 336 h (fasted) after ingestion of a single bolus of 45 mg uniformly labeled (13)C18:2n-6 dissolved in 8 g olive oil. In three subjects, breath was also sampled and CO(2) production measured every hour during the first 12 h. Subjects consumed their habitual diets. Plasma (13)C-enrichments were measured by GC-C-IRMS and fatty acid compositions by GC/FID. SETTING: Maastricht University, Department of Human Biology. SUBJECTS: Three men and three women, recruited by local advertisement. RESULTS: The tracer/tracee ratio (TTR) of C18:2n-6 in plasma total lipids was already increased 5 h after tracer intake. The mean peak amount (+/-s.e.m) of (13)C18:2n-6 (3.4+/-0.8 mg; 7.6% of dose) was found after about 17 h, (13)C18:3n-6 (0.018+/-0.008 mg; 0.04% of dose) after 7-48 h, and (13)C20:3n-6 (0.028+/-0.011 mg; 0.06% of dose) after 48-336 h. Time to peak TTRs of C20:4n-6 varied between subjects and were on average 0.022+/-0.006 mg (0.05% of dose). The proportion of (13)C18:2n-6 recovered in breath after 12 h ranged between 16.8 and 25.1%. CONCLUSIONS: These findings suggest that a single bolus of 45 mg U-(13)C18:2n-6 can be used to study the oxidation of (13)C18:2n-6. However, because of the low TTRs for C20:4n-6, a higher dose is recommended for studying the conversion of (13)C18:2n-6 into LCPs. In addition, since only about 35% of the tracer was found in plasma total lipids and as (13)CO(2) in breath, it might be necessary to study other accessible lipid fractions as well to study the overall conversion of linoleic acid.

Effects of dietary alpha-linolenic acid on the conversion and oxidation of 13C-alpha-linolenic acid.

The effects of a diet rich in alpha-linolenic acid vs. one rich in oleic acid on the oxidation of uniformly labeled 13C-alpha-linolenic acid and its conversion into longer-chain polyunsaturates (LCP) were investigated in vivo in healthy human subjects. Volunteers received a diet rich in oleic acid (n = 5) or a diet rich in alpha-linolenic acid (n = 7; 8.3 g/d) for 6 wk before and during the study. After 6 wk, subjects were given 45 mg of 13C-alpha-linolenic acid dissolved in olive oil. Blood samples were collected at t = 0, 5, 11, 24, 96, and 336 h. Breath was sampled and CO2 production was measured each hour for the first 12 h. The mean (+/- SEM) maximal absolute amount of 13C-eicosapentaenoic acid (EPA) in plasma total lipids was 0.04 +/- 0.01 mg in the alpha-linolenic acid group, which was significantly lower (P = 0.01) than the amount of 0.12 +/- 0.03 mg 13C-EPA in the oleic acid group. Amounts of 13C-docosapentaenoic acid (DPA) and 13C-docosahexaenoic acid (DHA) tended to be lower as well. The mean proportion of labeled alpha-linolenic acid (ALA) recovered as 13CO2 in breath after 12 h was 20.4% in the ALA and 15.7% in the oleic acid group, which was not significantly different (P = 0.12). The cumulative recovery of 13C from 13C-ALA in breath during the first 12 h was negatively correlated with the maximal amounts of plasma 13C-EPA (r = -0.58, P = 0.047) and 13C-DPA (r = -0.63, P = 0.027), but not of 13C-DHA (r = -0.49, P = 0.108). In conclusion, conversion of 13C-ALA into its LCP may be decreased on diets rich in ALA, while oxidation of 13C-ALA is negatively correlated with its conversion into LCP. In a few pilot samples, low 13C enrichments of n-3 LCP were observed in a diet rich in EPA/DHA as compared to oleic acid.

Effects of point mutations in the readthrough domain of the beet western yellows virus minor capsid protein on virus accumulation in planta and on transmission by aphids.

Point mutations were introduced into or near five conserved sequence motifs of the readthrough domain of the beet western yellows virus minor capsid protein P74. The mutant virus was tested for its ability to accumulate efficiently in agroinfected plants and to be transmitted by its aphid vector, Myzus persicae. The stability of the mutants in the agroinfected and aphid-infected plants was followed by sequence analysis of the progeny virus. Only the mutation Y201D was found to strongly inhibit virus accumulation in planta following agroinfection, but high accumulation levels were restored by reversion or pseudoreversion at this site. Four of the five mutants were poorly aphid transmissible, but in three cases successful transmission was restored by pseudoreversion or second-site mutations. The same second-site mutations in the nonconserved motif PVT(32-34) were shown to compensate for two distinct primary mutations (R24A and E59A/D60A), one on each side of the PVT sequence. In the latter case, a second-site mutation in the PVT motif restored the ability of the virus to move from the hemocoel through the accessory salivary gland following microinjection of mutant virus into the aphid hemocoel but did not permit virus movement across the epithelium separating the intestine from the hemocoel. Successful movement of the mutant virus across both barriers was accompanied by conversion of A59 to E or T, indicating that distinct features of the readthrough domain in this region operate at different stages of the transmission process.

Effects of mutations in the beet western yellows virus readthrough protein on its expression and packaging and on virus accumulation, symptoms, and aphid transmission.

Virions of beet western yellows luteovirus contain a major capsid protein (P22.5) and a minor readthrough protein (P74), produced by translational readthrough of the major capsid protein sequence into the neighboring open reading frame, which encodes the readthrough domain (RTD). The RTD contains determinants required for efficient virus accumulation in agroinfected plants and for aphid transmission. The C-terminal halves of the RTD are not well conserved among luteoviruses but the N-terminal halves contain many conserved sequence motifs, including a proline-rich sequence separating the rest of the RTD from the sequence corresponding to the major coat protein. To map different biological functions to these regions, short in-frame deletions were introduced at different sites in the RTD and the mutant genomes were transmitted to protoplasts as transcripts and to Nicotiana clevelandii by agroinfection. Deletions in the nonconserved portion of the RTD did not block aphid transmission but had a moderate inhibitory effect on virus accumulation in plants and abolished symptoms. Deletion of the proline tract and the junction between the conserved and nonconserved regions inhibited readthrough protein accumulation in protoplasts by at least 10-fold. The mutants accumulated small amounts of virus in plants, did not induce symptoms, and were nontransmissible by aphids using agroinfected plants, extracts of infected protoplasts, or purified virus as a source of inoculum. Other deletions in the conserved portion of the RTD did not markedly diminish readthrough protein accumulation but abolished its incorporation into virions. These mutants accumulated to low levels in agroinfected plants and elicited symptoms, but could not be aphid-transmitted. A preliminary map has been produced mapping these functions to different parts of the RTD.

Essential fatty acid status in neonates after fish-oil supplementation during late pregnancy.

Healthy pregnant women (n 23) were supplemented with fish-oil capsules (2.7 g n-3 polyunsaturated fatty acids/d) from the 30th week of gestation until delivery. Subjects in a control group were either supplemented with olive-oil capsules (4 g/d, n 6) or received no supplementation (n 10). Fatty acid compositions of the phospholipids isolated from umbilical plasma and umbilical arterial and venous vessel walls were determined. Fatty acid compositions of maternal venous plasma phospholipids were determined as well. Maternal plasma phospholipids of the fish-oil-supplemented group contained more n-3 fatty acids and less n-6 fatty acids. Moreover, the amounts of the essential fatty acid deficiency markers Mead acid (20:3n-9) and Osbond acid (22:5n-6) were significantly lower. The extra amount of n-3 fatty acids consumed by the mothers resulted in higher contents of n-3 fatty acids, and of docosahexaenoic acid (22:6n-3) in particular, in the phospholipids of umbilical plasma and vessel walls. It is, indeed, possible to interfere with the docosahexaenoic acid status at birth: children born to mothers supplemented with fish oil in the last trimester of pregnancy start with a better docosahexaenoic acid status at birth, which may be beneficial to neonatal neurodevelopment.

Effects of dietary fatty acids on signal transduction and membrane cholesterol content in rat platelets.

Previously, we have reported that dietary fatty acids can modify the thromboxane A2-dependent activation of rat platelets. Here, we present evidence that this dietary effect is part of a more general effect on platelet signal transduction, putatively involving structural changes in the platelet membranes. Four experiments were performed, where Wistar rats were fed with a high-fat diet enriched in either saturated, n-6 polyunsaturated or n-3 polyunsaturated fatty acids, or with a low-fat diet enriched in n-6 polyunsaturated fatty acids. The type of diet hardly influenced mean number of double bonds in the major platelet phospholipids. Platelet membranes from the rats fed with the saturated-fat diet had phospholipids with relatively high levels of arachidonate, but were low in cholesterol/phospholipid ratio. When compared to this diet group, platelets from other groups had an arachidonate content that was 21 to 47% lower and a cholesterol/phospholipid ratio 3 to 5% higher. The saturated-fat diet resulted in platelets that, in general, were less responsive to agonists than the platelets from other groups: with thrombin, collagen and thromboxane A2 analogue U46619, both early (shape change and phospholipase C-dependent rise in [Ca2+]i) and late (exocytosis and aggregation) responses were relatively low. However, platelet activation evoked by ADP was not influenced by diet type. When the cholesterol content of rat platelets was modified in vitro, it appeared that the early and late responses to thrombin and U46619 increased with the cholesterol/phospholipid ratio. Taken together, these results suggest that in rat platelets (i) the membrane cholesterol/phospholipid ratio can be modulated by a diet rich in saturated fatty acids, explaining, at least in part, the dietary effect on phospholipase C-mediated platelet activation, and (ii) relatively small changes in cholesterol content can have a more profound effect on platelet activation than substantial changes in arachidonate level.

Essential fatty acid status of neonates born to Inuit mothers: comparison with Caucasian neonates and effect of diet.

Fatty acid compositions were determined of phospholipids isolated from venous cord plasma and from the walls of umbilical arteries and veins, collected from healthy, a terme, Inuit and Caucasian (Dutch) neonates. The Inuit fatty acid profiles were characterized by a lower essential fatty acid (EFA) status, with higher levels of monounsaturated fatty acids, of Mead acid [20:3(n-9)] and its direct elongation product, and with lower amounts of the longer chain (greater than or equal to 20 carbon atoms), highly unsaturated (greater than or equal to 4 double bonds) fatty acids of both the (n-3) and (n-6) families. Levels of linoleic- and dihomo-gamma-linolenic acids were higher in Inuit as compared to Caucasian neonates, which suggests a low activity of the delta-5-desaturase in the Inuit. Within the Inuit group, a higher intake of marine food was associated with a better neonatal (n-3) status. Although the differences between Inuit and Caucasian neonates may be of genetic rather than of dietary origin, the results imply that dietary long-chain (n-3) or (n-6) fatty acids may be particularly important during pregnancy in Inuit mothers. Further studies are indicated with respect to the EFA content of the habitual Inuit diet and levels of delta-5-desaturase activity in the Inuit.

Ovarian function in premenopausal women affected by breast cancer: the measurement of glucuronoconjugate metabolites of 17 beta-estradiol and progesterone throughout one entire menstrual cycle.

For many years, hypersecretion of estrogens has been suspected of being one of the major risk factors of breast cancer for premenopausal women. Seventeen premenopausal women, who had undergone lumpectomy because of breast cancer (T1a No Mo) 3 yr before entering the study, were compared to 9 normal women of similar age, parity and body weight. A chemiluminescent method was used for the determination of estrone-3-glucuronide (E1-3G) and pregnanediol-3-glucuronide (Pd-3G) in early morning urine samples collected for an entire menstrual cycle of each of the 26 subjects. During the follicular phase, no significant differences in E1-3G and/or Pd-3G excretion were found between the two groups. During the luteal phase the E1-3G/Pd-3G ratio in the early, middle and late luteal phase had significantly increased in the women with breast cancer, in spite of normal Pd-3G excretion. Therefore, the measurement of glucuronoconjugate metabolites of ovarian hormones in overnight urine might be conveniently applied to the study of ovarian function in subjects with breast cancer. Furthermore, the results of this study may indicate that an estrogen/progesterone imbalance is an additional risk factor for the premenopausal breast cancer patient.

Fatty acid composition of umbilical arteries and veins: possible implications for the fetal EFA-status.

Fatty acid compositions were determined of phospholipids, isolated from umbilical arteries and veins, obtained from Dutch neonates after vaginal delivery, terminating normal pregnancy. The fatty acid profiles of the cord vessels were characterized by the absence of eicosapentaenoic (timnodonic) acid, a low (2-3%) content of linoleic acid and reasonable amounts of arachidonic acid (10-15%) and docosahexaenoic (cervonic) acid (3-5%). Significant amounts of Mead acid (1-4%) and its direct elongation product (0.5-2%) were also observed. In each cord, the efferent blood vessels contained significantly more Mead acid and other fatty acids of the oleic acid (n-9) family and less fatty acids of the linoleic (n-6) and linolenic (n-3) families than the afferent blood vessel. This indicates that the essential fatty acid (EFA) status of 'downstream' neonatal tissue may be marginal. No signs of EFA-deficiency were observed in endothelial and smooth muscle cells in culture, or in blood vessels from adults. In all cords 22:5(n-6) was significantly higher in the artery compared to the vein, whereas for all other (n-6) fatty acids this difference was negative. Since the synthesis of 22:5(n-6) is known to be stimulated when the required amount of cervonic acid, 22:6(n-3), is too low, our observations also suggest that the cervonic acid status of the neonates investigated was not optimal. Further studies are in progress to relate these findings to maternal EFA status and complications of pregnancy.

Preparation of the multienzyme system gramicidin S-synthetase 2 with an aqueous three-phase system.

The distribution of gramicidin S-synthetase activity from disrupted cells suspended in aqueous two- and three-phase systems was investigated. An optimized three-phase system containing 5% dextran, 8% Ficoll, 11% PEG and 6.7% disrupted cells was found to be effective in extracting gramicidin S-synthetase activity. The activity yield achieved was higher in comparison to other preparation methods, and the subsequent purification steps were greatly facilitated. The time needed for the preparation of the labile gramicidin S-synthetase was considerably reduced. The combination of the aqueous phase extraction with chromatographic methods yielded 19 mg gramicidin S-synthetase 2 in essentially pure form from 30 g (wet weight) of cells.

A sodium dodecyl sulfate-gradient gel electrophoresis system that separates polypeptides in the molecular weight range of 1500 to 100,000.

A sodium dodecyl sulfate-polyacrylamide gradient gel system is described. It combines a linear gradient in polyacrylamide from 10.2 to 30.2% in the separating gel and the discontinuous ammediol/glycine buffer system suggested by Bury (A. F. Bury, 1981, J. Chromatogr. 213, 491-500). This urea-free electrophoretic system provides high resolution and clean separation patterns of proteins and polypeptides with molecular weights from 1500 to 100,000. It is especially suited for studying complex mixtures of proteins and proteolytic fragments, in particular with regard to immunoelectrotransfer blot techniques.

Early detection of breast cancer recurrences through periodic follow-up--is it useless?

The authors report on a multicentric consecutive series of 1120 breast cancer first recurrences. Cases detected as subjectively asymptomatic thanks to periodic follow-up examinations are compared to cases detected as symptomatic. The relapse-free interval from primary treatment was shorter for asymptomatic recurrences, confirming that an earlier diagnosis was achieved in these cases. In spite of this diagnostic anticipation, median and actuarial survival from primary treatment did not differ when asymptomatic recurrences were compared to symptomatic recurrences. The study results did not show any prognostic impact of periodic follow-up in breast cancer and urge for prospective controlled studies on this diffuse and expensive practice.

Recombinant plasmids with genes for the biosynthesis of alkaline phosphatase of Escherichia coli.

Restriction maps of several recombinant plasmids representing a section of the E. coli K12 chromosome 35,000 bp in size with the genes phoA, proC and phoB were prepared. The orientation of phoA and the exact position of its N-terminal end on this map were determined by identifying a subfragment which carried the phoA promoter and by determining the nucleotide sequence of a 160 bp portion of this subfragment comprising the codons for the N-terminal end of pre-alkaline phosphatase. From this DNA sequence the leader sequence of alkaline phosphatase which consists of 21 amino acids was derived.