Prophylactic Bilateral Salpingo-oophorectomy in BRCA2 Mutation with Incidental Finding of Serous Tubal Intraepithelial Carcinoma (STIC) and Subsequent Diagnosis of Primary Peritoneal Carcinoma (PPC): A Case Report and Review of Current Literature.

A major effort to prevent serous cancer in genetically susceptible women with breast cancer susceptibility gene (BRCA) mutations has recently introduced the practice of risk-reducing prophylactic salpingo-oophorectomy. A small number of those who undergo prophylactic salpingo-oophorectomy will be found to have occult carcinomas. The majority of these appear to originate in the fallopian tube, reinforcing the theory that a significant proportion of high-grade serous carcinoma pelvic tumours have a fimbrial origin. In addition to this, histopathological and molecular biological characteristics suggest that among other serous carcinomas, fallopian tube serous carcinoma and primary peritoneal serous carcinoma really represent one entity. We present a case with breast cancer susceptibility gene 2 (BRCA2) mutation that was found to have serous tubal intraepithelial carcinoma (STIC) following prophylactic salpingo-oophorectomy. Subsequently, she was diagnosed with advanced primary peritoneal carcinoma. This prompted our team to reflect upon the case, review the current literature and recommend a rigorous preoperative assessment and meticulous intraoperative examination for prevention and early detection of high grade serous pelvic carcinomas.

Squamous cell carcinoma arising from a vulval epidermal cyst.

Malignant transformation of epidermal cyst is rare, with most cases arising in the head and neck. Here, we report a case of squamous cell carcinoma arising from a longstanding epidermal cyst in the right vulva of a 65-year-old woman. The patient presented with a sudden increase in size of the cyst over 6 weeks to the size of a golf ball. Excisional biopsy and histology indicated moderately differentiated squamous cell carcinoma within an epidermal cyst (FIGO stage IB). Strong p16 immunopositivity was also demonstrated and the potential significance of this is discussed. Limited right hemivulvectomy and right groin node dissection were performed.

Pregnancy outcome after electrosurgical cervical cone biopsy using Fischer cone biopsy excisor.

OBJECTIVE: To evaluate the pregnancy outcomes of all patients who underwent electrosurgical cone biopsy of the cervix between January 2000 and December 2011 and subsequently became pregnant. STUDY DESIGN: Retrospective cohort study. SETTINGS: District General Hospital in the North East of England. METHODS: Patients were identified from the local colposcopy electronic data, Hospital Episode Statistics and Maternity electronic data. Data were collected on a pro forma with two sections: (1) treatment section and (2) pregnancy section. In the treatment section, year and indication for treatment, volume of cervix removed, histological results and marginal status of specimen were documented. In the pregnancy section, time interval between treatment and pregnancy, pre-treatment obstetric history, cervical length measurements, cervical suture, gestation and mode of delivery and neonatal outcome were documented. Data were analysed using SPSS. RESULTS: 25 women achieved 47 pregnancies after electrosurgical cone biopsy treatment. Most common indication for cone biopsy was glandular neoplasia accounting for nearly half of the procedures; 21.2% of pregnancies ended in first-trimester miscarriages. The preterm delivery rate (<37 weeks) was 19.4%. Volume of cervix excised was significantly greater in women who delivered preterm compared to women who delivered at term (p = 0.028). The rate of preterm delivery was significantly higher in post treatment pregnancies when compared to pregnancies before treatment in the same women (p = 0.02). The preterm delivery in post-treatment pregnancies was not related to the time interval between treatment and pregnancy (p = 0.54). There was no significant difference in miscarriage rates in pre- and post-treatment pregnancies (p = 0.98). CONCLUSION: Electrosurgical cone biopsy of cervix is associated with increased risk of preterm labour that is related to the volume of cervix excised.

Liquid chromatography-tandem mass spectrometry method for the estimation of adefovir in human plasma: Application to a pharmacokinetic study.

An analytical method based on solid phase extraction was developed and validated for analysis of adefovir in human plasma. Adefovir-d4 was used as an internal standard and Synergi MAX RP80A (150 mm×4.6 mm, 4 µm) column provided the desired chromatographic separation of compounds followed by detection with mass spectrometry. The method used simple isocratic chromatographic condition and mass spectrometric detection in the positive ionization mode. The calibration curves were linear over the range of 0.50-42.47 ng/mL with the lower limit of quantitation validated at 0.50 ng/mL. Matrix effect was assessed by post-column infusion experiment to monitor phospholipids and post-extraction addition experiment was performed. The degree of matrix effect for adefovir was determined as 7.5% and ion-enhancement in five different lots of human plasma was 7.1% and had no impact on study samples analysis with 4.5 min run time. The intra- and inter-day precision values were within 7.7% and 7.8%, respectively, for adefovir at the lower limit of quantification level. Validated bioanalytical method was successfully applied to clinical sample analysis.

Metaxalone estimation in biological matrix using high-throughput LC-MS/MS bioanalytical method.

Metaxalone is a skeletal muscle relaxant, an approved drug for pain relief. Published bioanalytical methods lacked detailed stability evaluation in blood and plasma. An accurate, precise, high-throughput tandem mass spectroscopic method has been developed and validated. Following solid phase extraction (SPE), metaxalone and the internal standard metaxalone-d(3) were extracted from an aliquot of 200 µL of human plasma. Chromatographic separation achieved on an Ascentis Express C18 column (50 mm × 4.6 mm i.d., 2.7 µm particle size) with mobile phase is a mixture of 10mM ammonium acetate buffer (pH 4.5)-methanol-acetonitrile (20:50:30, v/v/v), at an isocratic flow rate of 0.7 mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode via electrospray ionization (ESI) source. The mass transitions of metaxalone and metaxalone-d(3) were m/z 222.3→161.2 and m/z 225.3→163.3, respectively. The linear calibration curves were obtained in the concentration range of 0.105-10.081 µg/mL (r(2)≥0.99) with a lower limit of quantification (LLOQ) of 0.105 µg/mL. The intra- and inter-day precisions and relative error were all within 6%. Despite achieving high mean recovery (>78%), no interference peaks or matrix effects were observed. Detailed stability exercises including drug stability in blood, hemolyzed, lipemic and normal plasma were conducted to extend the method applicability in vast majority of clinical studies using 800 mg metaxalone extended release oral dosage form.

ESI-MS/MS stability-indicating bioanalytical method development and validation for simultaneous estimation of donepezil, 5-desmethyl donepezil and 6-desmethyl donepezil in human plasma.

A bioanalytical method was developed and validated to estimate donepezil, 6-desmethyl donepezil and 5-desmethyl donepezil simultaneously in human plasma using galantamine as an internal standard (IS). The chromatographic separation was achieved on a reverse-phase XTerra RP (150 × 4.6 mm, 5 µm) column without affecting recovery (mean recovery > 60% with CV < 10%) for all analytes. ESI-MS/MS multiple reaction monitoring in positive polarity was used to detect mass pairs for donepezil (m/z 380.3 → 91.3), 6-desmethyl donepezil (m/z 366.4 → 91.3), 5-desmethyl donepezil (m/z 366.4 → 91.3) and galantamine m/z (288.1 → 213.0). The linearity was established over a dynamic range of 0.339-51.870, 0.100-15.380 and 0.103-15.763 ng/mL for donepezil, 6-desmethyl donepezil and 5-desmethyl donepezil, respectively. The current method shows that minimal conversion of labile metabolites to parent donepezil in plasma as stability was successfully achieved for 211 days at -15 °C storage temperature. The method was successfully applied to a clinical study after administration of 10 mg donepezil tablets to healthy male Indian volunteers.

Rational design for variability minimization in bioanalytical method validation: illustration with LC-MS/MS assay method for terbinafine estimation in human plasma.

Terbinafine, a widely used antifungal drug, is a challenging molecule for quantitative bioanalysis due to certain factors contributing assay variability. Despite previous attempts at human plasma determination of terbinafine, exhaustive stability of the drug or an internal standard was lacking. Internal standard stability with negligible variation throughout the analysis is an indicator of a reliable bioanalytical method as the majority of LC-MS/MS assays are based on analyte/IS response ratios for quantitation. A newly developed high-throughput simple LC-MS/MS method is described for human plasma determination of terbinafine using naftifine internal standard and eluting all compounds within 2 min. A solid-phase extraction of terbinafine achieving mean recovery of 84.3% (CV < 4%) without compromising sensitivity (limit of quantitation 5.11 ng/mL) or linearity (5.11-3014.19 ng/mL) is delineated in this paper. A heated nebulizer in positive multiple reaction monitoring mode was employed with transitions m/z 292.2 →141.1 and 288.2 →117.0 for terbinafine and naftifine, respectively, resulting in excellent chromatographic separation on a Hypurity Advance (50 x 4.6 mm, 5 µm) column. The developed method was successfully applied to clinical samples and for the first time demonstrated marked improved extraction efficiency and reliable long-term plasma stability results without any internal standard response variation during the entire course of study.