Automated piezoresponse force microscopy domain tracking during fast thermally stimulated phase transition in CuInP2S6.

Real-time tracking of dynamic nanoscale processes such as phase transitions by scanning probe microscopy is a challenging task, typically requiring extensive and laborious human supervision. Smart strategies to track specific regions of interest (ROI) in the system during such transformations in a fast and automated manner are necessary to study the evolution of the microscopic changes in such dynamic systems. In this work, we realize automated ROI tracking in piezoresponse force microscopy during a fast (≈0.8°C s-1) thermally stimulated ferroelectric-to-paraelectric phase transition in CuInP2S6. We use a combination of fast (1 frame per second) sparse scanning with compressed sensing image reconstruction and real-time offset correction via phase cross correlation. The applied methodology enablesin situfast and automated functional nanoscale characterization of a certain ROI during external stimulation that generates sample drift and changes local functionality.

A freeze-all strategy does not increase live birth rates in women of advanced reproductive age.

RESEARCH QUESTION: Does a freeze-all strategy improve live birth rates in women of different age groups? DESIGN: Retrospective cohort study of 1882 first embryo transfer cycles, performed between January 2013 and December 2015. Reproductive outcomes between fresh (FRESH) or frozen (FROZEN) embryo transfers were compared in patients stratified by age: < 35, between 35 and 38, or > 38 years. Student's t test for independent samples and χ2 analyses were used as needed. A multivariable logistic regression analysis was performed adjusting for age, triggering drug, number of retrieved oocytes, number of transferred embryos, and percentage of top-quality embryos. MAIN RESULTS AND THE ROLE OF CHANCE: Live birth rates (LBR) were significantly higher for FROZEN in the < 35 years group (43.7% vs 24%; p < 0.001). In both the 35-38 and > 38 years groups, LBR for FROZEN vs FRESH were not statistically different (30.9% in the FROZEN group vs 29.3% in the FRESH group, p = 0.70, and 19.8% in the FROZEN group vs 12.7% in the FRESH group, p = 0.07, respectively). The multivariate analysis found a significantly positive effect of performing FROZEN on LBR in the younger group (OR 2.46, 95% CI 1.31-4.62; p = 0.005) but had no impact in women between 35 and 38 years (OR 1.01, 95% CI 0.55-1.83; p = 0.98) or older (OR 0.96, 95% CI 0.43-2.13; p = 0.92). CONCLUSIONS: Performing a freeze-all strategy seems to result in better reproductive outcomes when compared with a fresh ET in women under 35 years, with no significant impact on older women.

ALWAYS ICSI? A SWOT analysis.

PURPOSE: Intracytroplasmatic sperm injection (ICSI) is a common procedure used to improve reproductive results, even among couples without male factor infertility. However, the evidence available is still uncertain on the possible advantages and deficiencies that this procedure may have in patients with no formal indication for ICSI. METHODS: A SWOT (strengths, weaknesses, opportunities, threats) analysis examines the possible advantages and deficiencies of performing ICSI in these patients with no formal indication. RESULTS: The evidence suggests that ICSI is not justified for non-male factor infertile couples requiring in vitro conception. One of the major strengths associated to the procedure is the virtual elimination of cases further complicated by total fertilization failure and a combination between IVF and ICSI on sibling oocytes has been advised in the literature. Greater technical difficulties, higher costs and performing an unnecessary invasive technique in some cases represent some of the weaknesses of the procedure, and questions regarding safety issues should not be ruled out. CONCLUSION: Despite the widespread use of ICSI in patients without a formal diagnosis of male factor infertility, evidence demonstrating its effectiveness in this population is still lacking. Additional large and well-designed randomized controlled trials are needed to clarify definitive indications for ICSI in non-male factor infertility.

Oral versus transdermal oestrogen delivery for endometrial preparation before embryo transfer: a prospective, comparative, randomized clinical trial.

RESEARCH QUESTION: To determine whether the transdermal route is equal or superior to the oral route, when preparing the endometrium with oestrogens for embryo transfer. DESIGN: Prospective, randomized controlled trial; 140 patients randomized; the pills group followed a protocol with oestradiol valerate pills and the patches group followed a protocol with oestradiol hemihydrate patches. The primary variable was endometrial thickness on day 10 ± 1 of treatment. Secondary variables were endometrial thickness on day 15 ± 1 of treatment, patient satisfaction, plasma levels of oestradiol, rates of pregnancy, miscarriage and delivery. Endometrial thickness was measured on day 10 ± 1 of the cycle, if the lining was 7 mm or less in thickness, another measurement was made on day 15 ± 1. Blood oestradiol levels were analysed on the day the endometrial lining was greater than 7 mm (day 10 ± 1 or day 15 ± 1). Patients completed a survey to evaluate comfort and side-effects. RESULTS: The patches group achieved significantly thicker endometrium by the first check-up on day 10 ± 1 (7.6 mm versus 7.0 mm; P = 0.026), with lower blood levels of oestradiol (159.2 pg/ml versus 237.1 pg/ml; P < 0.001) when the endometrial thickness was over 7mm. The pills group considered the treatment more comfortable, with less side-effects. No significant differences in the rates of pregnancy, miscarriage or live birth were found. CONCLUSIONS: Transdermal oestrogen treatment allows patients to reach a higher endometrial thickness after 10 days of treatment, with lower plasma levels of oestradiol, although it is not tolerated as well.

Hysteroscopic septum resection and reproductive medicine: A SWOT analysis.

Hysteroscopic septum resection in women with unfavourable reproductive and clinical outcomes has become common practice worldwide to improve reproductive results. No clear evidence on the possible advantages and drawbacks of this procedure has been published. In this opinion paper, based on a SWOT (strengths, weaknesses, opportunities, threats) analysis, the different aspects of this strategy are evaluated. Currently, no level 1 published evidence supports uterine resection in women with septate uterus. Clinical evidence from the studies analysed matches the more recent guidelines and suggests an improvement in reproductive outcomes after hysteroscopic resection of the septum, particularly in infertile women and women who have experienced recurrent miscarriages. In a patient with no history of infertility or prior pregnancy loss, it may be reasonable to consider septum incision after counselling about the potential risks and benefits of the procedure. Published clinical data in favour of the intervention, however, are based on studies with important methodological limitations. In this situation, the clinician and patient should reach an agreement together, based on the pros and cons of this intervention. Well-designed randomized controlled trials are required to confirm the clinical benefits and cost-effectiveness of this procedure.

There is no evidence that the time from egg retrieval to embryo transfer affects live birth rates in a freeze-all strategy.

STUDY QUESTION: Does the time from ovum pick-up (OPU) to frozen embryo transfer (FET) affect reproductive outcomes in a freeze-all strategy? SUMMARY ANSWER: Our study did not detect statistically significant differences between first and subsequent cycles, clinically relevant differences are not ruled out and further and larger studies are required. WHAT IS KNOWN ALREADY: Following controlled ovarian hyperstimulation (COH) delaying FET until the endometrium has returned to an optimal pre-stimulation state may have a significant emotional impact on patients, which adds to the stress and anxiety accompanying a standard IVF cycle. Currently there is no agreement on the best time to perform a FET after a freeze-all cycle in order to maximize reproductive outcomes for the patient. STUDY DESIGN, SIZE, DURATION: Retrospective cohort study of 512 freeze-all cycles, performed between January 2012 and December 2014. COH was performed by either a GnRH antagonist (n = 397) or a long GnRH agonist protocol (n = 115). Ovulation was triggered using either a GnRH agonist (n = 258) or hCG (n = 254). Endometrial preparation was performed in an artificial cycle by either oral (n = 238) or transdermal (n = 274) oestrogen. Differences were considered significant if P < 0.05. PARTICIPANTS/MATERIALS, SETTING, METHODS: Reproductive outcomes between FETs which took place either within the first menstrual cycle following OPU (Cycle 1; n = 263) or afterwards (Cycle ≥2; n = 249) were compared. Student's t-test for independent samples, Mann-Whitney U-test and Chi-square analysis were used where appropriate. A multivariable logistic regression analysis was performed adjusting for maternal age, drug used for ovulation trigger, number of retrieved oocytes, number of embryos obtained, day of embryonic development at transfer, number of embryos transferred and type of endometrial preparation. Differences were considered significant if P < 0.05. MAIN RESULTS AND THE ROLE OF CHANCE: Live birth rate (LBR) was significantly higher in FET performed during Cycle 1 vs Cycle ≥2 (37.6% vs 27.3%, respectively; P = 0.01) before adjusting for confounding factors. We found no difference for biochemical pregnancy (49.8% vs 43.8%; P = 0.17), clinical pregnancy (44.1% vs 36.1%; P = 0.07) or pregnancy loss (11.8% vs 16.1%; P = 0.16). A multivariable analysis found no impact of timing of elective FET on LBR (odds ratio, OR 0.73; 95% CI 0.49-1.08). The impact remained not significant after adjusting for number of retrieved oocytes, drug used for ovulation trigger (hCG vs GnRH agonist) and reason for cryopreservation. The factors that significantly affected LBR were: maternal age in both age categories (women between 35 and 40 years vs women below 35 years, OR 0.63, 95% CI 0.4-0.95; and women over 40 years vs women below 35 years, OR 0.34, 95% CI 0.2-0.7), day of embryonic development at transfer (day +4 vs +3; OR 1.7, 95% CI 1.1-2.8) and number of transferred embryos (OR 2.2, 95% CI 1.4-3.3) and oestrogen used for endometrial preparation (transdermal vs oral; OR 0.62, 95% CI 0.4-0.9). LIMITATIONS REASONS FOR CAUTION: The main limitation of our study is its retrospective nature. Although we adjusted our statistical analysis for a number of known and suspected confounders, we cannot exclude the possibility of residual confounding factors. WIDER IMPLICATIONS OF THE FINDINGS: According to our results, clinicians might not need to wait more than one menstrual cycle before performing FET. This allows us to reduce unnecessary delays in FET, without compromising reproductive outcomes. STUDY FUNDING/COMPETING INTERESTS: No funding was sought for this study. Authors declare no competing interests. TRIAL REGISTRATION NUMBER: NA.

Obesity decreases the oxidant stress induced by tobacco smoke in a rat model.

Obesity and emphysema are associated with low-grade systemic inflammation and oxidant stress. Assuming that the oxidant stress induced by emphysema would be decreased by obesity, we analyzed the oxidant/antioxidant state in a rat model combining both diseases simultaneously. Obesity was induced using sucrose, while emphysema by exposure to tobacco smoke. End-points evaluated were: body weight, abdominal fat, plasma dyslipidemia and malondialdehyde (MDA), insulin and glucose AUC, activities of Mn-superoxide dismutase (Mn-SOD), glutathione reductase (GR), glutathione transferase (GST) and glutathione peroxidase (GPx); lung MnSOD and 3-nitrotyrosine (3-NT) immunostaining, and expression of αV and ß6 integrin subunits. In rats with obesity, the body weight, abdominal fat, plasma triglyceride levels, glucose AUC, insulin levels, GST activity, and αV and ß6 integrin expressions were amplified. The rats with emphysema had lower values of body weight, abdominal fat, plasma insulin, triglycerides and glucose AUC but higher values of plasma MDA, GPx activity, and the lung expression of the αV and ß6 integrins. The combination of obesity and emphysema compared to either condition alone led to diminished body weight, abdominal fat, plasma insulin MDA levels, GPx and GST activities, and αV and ß6 integrin expressions; these parameters were all previously increased by obesity. Immunostaining for MnSOD augmented in all experimental groups, but the staining for 3-NT only increased in rats treated with tobacco alone or combined with sucrose. Results showed that obesity reduces oxidant stress and integrin expression, increasing antioxidant enzyme activities; these changes seem to partly contribute to a protective mechanism of obesity against emphysema development.

Renin is an angiotensin-independent profibrotic mediator: role in pulmonary fibrosis.

The pathogenesis of idiopathic pulmonary fibrosis (IPF) is probably the result of interplay between cytokines/chemokines and growth factors. The renin-angiotensin (Ang) system is involved, although its profibrotic effect is attributed to Ang II. However, recent studies suggest that renin, through a specific receptor, is implicated in fibrogenesis. In this study, the expression of renin and renin receptor was examined in normal and IPF lungs and fibroblasts. Normal human lung fibroblasts were stimulated with renin or transfected with renin small interfering RNA (siRNA), and the expression of transforming growth factor (TGF)-ß1 and α-1-type I collagen was analysed. Normal lungs and lung fibroblasts expressed renin, which was strongly upregulated in IPF lungs and fibroblasts (∼10-fold increase; p<0.05). Immunocytochemistry showed intense renin staining in IPF fibroblasts. Renin-stimulated lung fibroblasts displayed an increase in the expression of TGF-ß1 (mean ± sd 1.8 × 10(3) ± 0.2 × 10(3) versus 1.2 × 10(3)± 0.3 × 10(3) mRNA copies per 18S ribosomal RNA; p<0.01) and collagen (5.93 × 10(2)± 0.66 × 10(2) versus 3.28 × 10(2) ± 0.5 × 10(2); p<0.01), while knocking down renin expression using siRNA provoked a strong decrease of both molecules. These effects were independent of Ang II, since neither losartan nor captopril decreased these effects. Renin also decreased matrix metalloprotease-1 expression and induced TGF-ß1 activation (163 ± 34 versus 110 ± 15 pg active TGF-ß1 per mg total protein). These findings highlight the possible role of renin as an Ang II-independent profibrotic factor in lung fibrosis.

Analysis of pain perception associated with urodynamic testing in children over 3 years old. / Análisis de la percepción del dolor asociado al estudio urodinámico en niños mayores de tres años.

OBJECTIVE: To measure the tolerance of urodynamic testing (UDT) in the pediatric patient by means of the Visual Analog Scale (VAS). To analyze which clinical and UDT-related variables influence pain perception. MATERIAL AND METHODS: Cross-sectional study of 139 pediatric patients undergoing UDT (December 2013 - May 2018). INCLUSION CRITERIA: understanding and expressing their experience after UDT (preschool and school age). No adolescents were included. Measurement instrument Visual Analog Scale (0-10). Other clinical and UDT-associated variables were obtained. STATISTICAL ANALYSIS: Mann-Whitney U test, Kruskal Wallis test. Spearman's rank correlation analysis (rs). Multivariate analysis through ordinal logistic regression. Significance p < 0.05. RESULTS: Mean age 7.7 years (SD 2.4), median VAS score, 2 (2-6). In 41% (n = 57), the score was ≥ 4 (moderate pain). Multivariate analysis. Explanatory variables for obtaining a high VAS score: high APUDT score (identifying patient anxiety prior to UDT), sensory-motor alteration in the lower limbs, difficult bladder catheterization and the appearance of pain during the filling phase. Age and duration of the UDT have not influenced the VAS score. CONCLUSIONS: Although the UDT has resulted in 40% of the pediatric patients in our study expressing discomfort or pain, it is a well-tolerated test. The variables that have influenced on pain perception were patient's anxiety prior to UDT, a sensory-motor alteration located in the lumbosacral metameres, difficult bladder catheterization and the appearance of pain during bladder filling.

Out-of-protocol concurrent use of cisplatin and radiation therapy in locally advanced cervical cancer: feasibility and survival.

PURPOSE OF INVESTIGATION: We assessed the feasibility, response rates, and overall survival of patients with locally advanced cervical cancer treated with cisplatin-based chemotherapy during radiation therapy on an out-of-protocol basis. METHODS: Sixty-nine consecutive newly diagnosed untreated patients with locally advanced cervical cancer who received chemoradiation between 1999 and 2003 were retrospectively reviewed. Treatment consisted in external beam radiation followed by one 137-cessium intracavitary application. Cisplatin was administered for six weeks during external beam radiation. RESULTS: Treatment was well tolerated, although 52 patients presented some degree of acute adverse toxicity (gastrointestinal 65%, hematological 48%, genitourinary 10%). The 3-year survival rate was 61.8% (95% CI 54.5-69.0), with a mean 41.8 months (95% CI 35.7-48.3). Overall survival after adjusting by FIGO Stage IB2-IIA and IIB-IVA was 73.9% and 50%, respectively (p = 0.1839). Overall survival according to Stages IB2-IIb and III-IVA was 74.8% and 34.9%, respectively (P = 0.0376). CONCLUSION: In patients with locally advanced cervical cancer, adding a weekly regimen of cisplatin to standard pelvic radiation in an out-of-protocol basis is feasible, effective, and showed no unexpected toxicity.

Discovery, characterization and validation of single nucleotide polymorphisms within 206 bovine genes that may be considered as candidate genes for beef production and quality.

A large number of putative single nucleotide polymorphisms (SNPs) have been identified from the bovine genome-sequencing project. However, few of these have been validated and many will turn out to be sequencing artefacts or have low minor allele frequencies. In addition, there is little information available on SNPs within coding regions, which are likely to be responsible for phenotypic variation. Therefore, additional SNP discovery is necessary to identify and validate polymorphisms both in specific genes and genome-wide. Sequence-tagged sites within 286 genes were resequenced from a panel of animals representing a wide range of European cattle breeds. For 80 genes, no polymorphisms were identified, and 672 putative SNPs were identified within 206 genes. Fifteen European cattle breeds (436 individuals plus available parents) were genotyped with these putative SNPs, and 389 SNPs were confirmed to have minor allele frequencies above 10%. The genes containing SNPs were localized on chromosomes by radiation hybrid mapping and on the bovine genome sequence by Blast. Flanking microsatellite loci were identified, to facilitate the alignment of the genes containing the SNPs in relation to mapped quantitative trait loci. Of the 672 putative SNPs discovered in this work, only 11 were found among the validated SNPs and 100 were found among the approximately 2.3 million putative SNPs currently in dbSNP. The genes studied in this work could be considered as candidates for traits associated with beef production and the SNPs reported will help to assess the role of the genes in the genetic control of muscle development and meat quality. The allele frequency data presented allows the general utility of the SNPs to be assessed.

Frequency-dependent force between ac-voltage-biased plates in electrolyte solutions.

We analyze the frequency dependence of the force between ac-voltage-biased plates in electrolyte solutions. To this end we solve analytically the Poisson-Nernst-Planck transport model in the dilute concentration and low voltage regime for a 1:1 symmetric electrolyte with blocking electrodes under a dc+ac applied voltage. The total force, which is the resultant of the electric and osmotic forces, shows a complex dependence on plate separation, frequency, ion concentration, and compact layer properties, different from that predicted from electrostatic current models or equivalent circuit models, due to the relevance of the osmotic force contribution in almost the whole range of frequencies. For the total dc force, we show that it decays at fixed ion concentration, linearly with plate separation for separations larger than a few times the Debye screening length. This linear dependence is due to the assumption about the conservation of the number of ions in the system. Moreover, the 1ω and 2ω ac harmonics of the total force show a broad peak at intermediate frequencies; it is centered at about the inverse of the charging time of the double layer capacitance, and covers the frequency range between the inverse of the diffusion time and the inverse of the electrolyte dielectric relaxation time. Finally, the 1ω ac harmonic component attains its high frequency asymptotic value at frequencies much higher than the inverse of the electrolyte dielectric relaxation time due to the very slow relaxation of the osmotic 1ω harmonic component at high frequencies. The derived analytical expressions for the total force remain valid up to voltages of the order of the thermal voltage, as has been assessed by means of numerical calculations. The numerical calculations are also used to explore the onset of higher force harmonics for larger applied voltages. Understanding the frequency dependence of the force acting on voltage-biased plates in electrolyte solutions can be of relevance for electrical actuation strategies in microelectromechanical systems and for the interpretation of some emerging electric scanning probe force microscopy techniques operating in electrolyte solutions.

Graphene oxide/titania photocatalytic ozonation of primidone in a visible LED photoreactor.

A graphene oxide-titania (GO/TiO2) composite was synthesized via sol-gel method, and studied in aqueous Primidone mineralization with ozone and LED visible light. The photocatalyst was characterized by different techniques (XRD, TEM, SBET, TGA, UV-vis diffuse reflectance spectroscopy). The band gap value decrease from 3.14 eV for bare TiO2 samples to 2.5 eV in GO/TiO2 composites clearly shows the interaction of GO with TiO2 structure. Approximately 20 mg L-1 of Primidone was removed in less than 20 min if ozone was applied, regardless of the presence or absence of light and catalyst. However, reactivity tests show a synergism effect between photocatalysis and ozonation for mineralization purposes. The combination of ozone and GO improved the activation of TiO2 under visible light. Process optimization led us to select a catalyst dosage of 0.25 g L-1, a light radiance of 359 W m-2 and a GO loading in the catalyst around 0.75%. At these conditions, with photocatalytic ozonation, the presence of GO in the catalyst improved mineralization up to 82% in 2 h compared to 70% reached with bare TiO2. Catalyst reusability shows no decrease of photocatalytic activity. Scavenger tests point to hydroxyl radicals as the main species responsible for Primidone removal.

[Mycophenolate mofetil in the treatment of lupus nephritis, in patients with failure, intolerance or relapses after treatment with steroids and cyclophosphamide]. / Micofenolato mofetil en el tratamiento de la nefritis lúpica en pacientes con fracaso, intolerancia o recidivas tras tratamiento con esteroides y ciclofosfamida.

Intravenous cyclophosphamide (IVCP) in combination with oral steroids (ST) is the most widely accepted therapy for severe lupus nephritis (LN); however, its side effects, lack of response and relapses, have led to other treatment alternatives. being sought. Mycophenolate mofetil (MMF) has been shown to be effective in these cases. We studied the course over 12 months of 28 patients with LN WHO class III(n=3), IV(n=22) or V(n=3), with 38,1 +/- 11,4 tears of age, proteinuria 4,2 +/- 2,6 g /24 hours and serum creatinine 1,4 +/- 0,8 mg/dL, who, after being initially treated with ST and IVCP, showed no response(n=21), frequent relapses(n=6), or adverse side effects(n=1). All patients were treated with MMF in doses of 1000 to 2000 mg/day combined with ST or cyclosporine for one year. Four patients withdrew from treatment before the end of the follow-up. None of the patients who completed the study showed changes in hematologic parameters. Creatinine and creatinine clearance remained stable. Resulted in a significant improvement; serum albumine (3 +/- 0,8 vs 3,9 +/- 0,5 g/dL) p<0.01, and decreased of proteinuria (4,2 +/- 2,6 vs 1,8 +/- 2,2 g/ 24 hours) p<0.05, complement fractions improvement significantly, C3 and CH50 p<0.05, C4 p<0.01. Antinuclear antibodies (ANA) and anti-DNA antibodies decreased significantly (p<0.05). During follow-up, a reduction in the ST dose was achieved: 18.3 +/- 10,5 vs 10,1 +/- 4,1 mg/24h (p<0.01). Three mild side effects related to MMF were observed and only 1 case required discontinuation of treatment. We concluded that MMF is a useful drug in the treatment and control of lupus nephritis, which also allows for a significant reduction in the dose of ST, with minimal side effects.

A comparison of raloxifene and calcium plus vitamin D on vaginal atrophy after discontinuation of long-standing postmenopausal hormone therapy in osteoporotic women. A randomized, masked-evaluator, one-year, prospective study.

OBJECTIVES: We assessed the effects of the discontinuation of long-standing transdermal estrogen replacement therapy (>4 years) and substitution of this treatment by calcium or raloxifene on the vaginal epithelium and climateric symptoms in a study population of osteoporotic women. METHODS: A total of 136 women (mean age 55.5 years) were randomized to calcium (500 mg elemental calcium, 400 IU Vitamin D3) (n=40), raloxifene (60 mg/day) (n=48), or estrogen patches (3.9 mg estradiol) and progesterone (100 mg/day) (n=48). Treatment was given for 1 year. The vaginal maturation value (VMV), serum estradiol levels, and climateric symptoms using a 12-item modification of the SF-36 quality of life questionnaire were evaluated at baseline and at 6 and 12 months. RESULTS: At 6 months, mean VMV decreased significantly (P<0.001) in the calcium (-51.8%) and raloxifene (-18.6%) groups compared with baseline and the estrogen/progesterone group. At 12 months, significant decreases of mean VMV in the calcium (-38.7%) and raloxifene groups (-32%) (P<0.001) were also observed. Serum estradiol levels and changes of VMV correlated significantly at 6 months (rho=0.361, P<0.01) and at 12 months (rho=0.269, P<0.035). A significantly higher number of patients complained of hot flushes and palpitations in the calcium and raloxifene groups than in the estrogen/progesterone group. Raloxifene-treated women reported a significantly higher number of adverse events at 6 months compared to the other treatment groups. CONCLUSIONS: Withdrawal and change of long-standing transdermal hormone replacement therapy by treatment with calcium or raloxifene resulted in worsening of vaginal atrophy assessed by the VMV, although it was not clinically perceived by the patients. However, increases in dyspareunia and urinary leaks were reported. Menopausal complaints related to vasomotor symptoms worsened in the calcium- and raloxifene-treated groups and persisted throughout the study period.

Trends in the use of emergency contraception: an epidemiological study in Barcelona, Spain (1994-2002).

We conducted a retrospective review of the medical records of women requesting emergency contraception (EC) at our emergency department over a 9-year period (1994-2002). EC accounted for 5.9% of all visits (n = 95,288) and increased from 1.26% in 1994 to 9.82% in 2002 (p < 0.001). Reasons for EC were condom problems in 79.5% of cases. EC was used only once by 93% of women. The mean daily number of visits was significantly higher in August (2.46), July (2.01) and September (2.02) than in other months (p < 0.05), and was more frequent on Sunday (3.26), Saturday (2.92) and Monday (2.05) compared to other week days (p < 0.001). New Year's Day and the St. John's Night registered the highest number of visits (mean of 17.2 and 11.7, respectively), with significant differences compared to the remaining days of the year (p < 0.001).

Determination of vitamin B(6) in cooked sausages.

A reverse-phase high-performance liquid chromatography (HPLC) method has been described for the determination of various active forms of vitamin B(6) in meat products. Different extracting agents were tested to solubilize fully the analyte for quantification. The best data were obtained by extracting the samples with 5% (w/v) metaphosphoric acid. Separation by HPLC was performed with fluorescence detection (excitation, 290 nm; emission, 395 nm), on a 10 cm x 0.46 cm i.d. Hypersil BDS C(18) 5 microm column using a mixture of 50 mM phosphate buffer (pH 3.2) and acetonitrile (99:1, v/v) as mobile phase. Precision of the method was 0.5% (within a day) and 4.3% (between days). The detection limits were 0.020 mg/100 g for pyridoxal and pyridoxamine, 0.017 mg/100 g for pyridoxamine phosphate, 0.500 mg/100 g for pyridoxal phosphate, and 0.033 mg/100 g for pyridoxol, with a signal-to-noise ratio of 3. The recovery ranged from 92.0 to 100.0%.

Determination of total riboflavin in cooked sausages.

A simple and rapid method for determining riboflavin content in cooked sausages by ion-pair reversed-phase liquid chromatography has been set up. Samples were subjected to acid and enzymatic hydrolysis. Sample extracts were directly chromatographed, avoiding purification and concentration treatment. Final determination was performed by high-performance liquid chromatography with fluorescence detector (excitation, 227 nm; emission, 520 nm), on a 25 cm x 4 mm i.d. Spherisorb ODS-2 cartridge using a mixture of 5 mM heptanesulfonic acid adjusted to pH 2.7 with phosphoric acid and acetonitrile (75:25, v/v) as mobile phase. Precision of the method was 1.3% (within a day) and 2.6% (between days). The detection limit was 0.015 mg/100 g. The recovery was >95%.