Analysis of free cisplatin in microdialysates and plasma ultrafiltrate by liquid chromatography-tandem mass spectrometry.

Cisplatin is a potent cytotoxic agent used in the treatment of various malignancies and exerts its antitumor effect through malignant cell DNA damage and apoptosis induction. Evaluation of systemic delivery of cisplatin is important in optimization of cisplatin treatment. However, accurate quantification of systemic cisplatin is challenging due to its various forms in circulation. This study aimed to develop a sensitive (LOQ < 0.1 µg/mL) and precise Ultra Performance Liquid Chromatography (UPLC) - Tandem Mass Spectrometry (MS/MS) method for quantifying free cisplatin in microdialysates and plasma. Furthermore the aim was to compare free cisplatin concentrations measured in standard plasma samples with those obtained from intravenous microdialysis catheters in a porcine model. The method developed utilizes dichloro(ethylenediamine)platinum(II) as an internal standard that co-elutes with cisplatin, ensuring precise correction for ion suppression/enhancement effects. The method was validated, demonstrating linearity up to 100 µg/mL and good intermediate precision (CV% < 6 %) in the range of 1.0-100 µg/mL, with an LOQ of 0.03 µg/mL. The pharmacokinetic parameters (AUC0-last, Cmax, T1/2, and Tmax) showed no significant differences between the two sampling methods. This validated LC-MS/MS method provides a reliable tool for quantifying systemic free cisplatin concentrations, facilitating future systemic and local pharmacokinetic evaluations for optimization of cisplatin-based cancer treatments.

Low attendance by non-native women to human papillomavirus vaccination and cervical cancer screening - A Danish nationwide register-based cohort study.

BACKGROUND: Cervical cancer is preventable through human papillomavirus vaccination and cervical cancer screening. However, possibly due to systemic, individual (e.g. low socio-economic staus) and socio-cultural barriers, it is likely that non-natives, especially non-westerns, are more prone to attend neither vaccination nor screening (combined non-attendance). This is disturbing as the non-native population in Denmark is predicted to rise to 21% by 2060. We aimed to investigate differences in combined non-attendance by nativity and region of origin, and to analyse the association between country of origin and combined non-attendance adjusted for socio-economic status. SETTING: 1.6.2007-31.12.2016 Denmark. METHODS: Logistic regression was performed to estimate crude and adjusted odds ratios with 95% confidence intervals for combined non-attendance. RESULTS: 170,158 women were included. Overall combined non-attendance was 11.8% [11.7-12.0]; 10.0% [9.8-10.1] for native women and 27.1% [26.4-27.7] for non-native women, with highest degrees among Middle-Eastern and North-Africans (30.1% [29.2-30.9]). Even when adjusted for socio-economics, women from Middle-East and North-Africa had substantially higher odds of combined non-attendance than natives (adj. OR = 7.5 [6.3-8.9] for Somali women). CONCLUSION: Denmark has a relatively low degree of combined non-attendance. However, cervical cancer preventive programmes seem to be better tailored to the needs of native women and do not appear to cater sufficiently to the needs of the fast-growing non-native populations, particularly not to the needs of Middle-Eastern and North African women. In order to secure more just cervical cancer prevention, future studies are recommended to develop tailored intervention sensitive to the need of non-native women.

Validity and reliability of State-Trait Anxiety Inventory in Danish women aged 45 years and older with abnormal cervical screening results.

BACKGROUND: State Trait Anxiety Inventory (STAI) scale was developed in the 1980's and has been widely used both in clinical settings and in research. However the Danish version of STAI has not been validated. The aim of this study was to assess the validity and reliability of STAI - state anxiety scale in Danish women aged 45 years and older with abnormal cervical cancer screening results. METHODS: Women ≥45 years referred with an abnormal cervical cytology and healthy volunteers (n = 12) underwent cognitive interview after completing STAI. Further, STAI was sent out in an electronic questionnaire to women (n = 109) seen at the gynecological department with abnormal cervical cancer screening test during 2018. Validity and reliability of STAI was evaluated according to the Consensus-based Standards for the selection of health Measurement Instruments (COSMIN) checklist by examining internal consistency, test-retest reliability, measurement error, floor and ceiling, construct validity and content validity. RESULTS: In the cognitive interviews the content validity was evaluated to be very good. The internal consistency of the scale was excellent with Cronbach's α = 0.93. Test-retest reliability was good with an intra-class correlation coefficient of 0.80 and the systematic difference between test-retest results was negligible. The construct validity was good. CONCLUSION: To our best knowledge, this is the first validation study of the Danish translation of STAI-state anxiety scale. This version of STAI demonstrates an acceptable reliability and validity when used in a gynecological setting.

Can adenocarcinoma in situ of the uterine cervix be treated safely by conisation in combination with endocervical curettage?

OBJECTIVE: To evaluate the prognostic value of endocervical curettage (ECC) after conisation in patients treated for adenocarcinoma in situ (AIS) of the uterine cervix. MATERIALS AND METHODS: Patients with AIS diagnosed between 1990 and 2010 and with a minimum of 1.5 years of follow-up were retrospectively identified using computerised clinical files. RESULTS: The authors identified 195 patients (median age 32 years) with a median follow-up of 6.4 years. ECC was performed in 165 patients. In 144 (87%) the initial ECC was normal. In 129 no recurrence was observed during follow-up (90%). A positive ECC was observed in 21. Thirteen patients had hysterectomies; six hysterectomies were normal. Eight patients treated conservatively developed no recurrent disease. Two patients with a positive ECC did not have a hysterectomy and developed recurrent disease. In patients with affected margins, 17% developed recurrent disease. CONCLUSION: ECC performed during initial conisation is a prognostic tool for the treatment ofAIS. Close follow-up is recommended in patients treated conservatively.

Amphiphilic polyanhydride nanoparticles stabilize Bacillus anthracis protective antigen.

Advancements toward an improved vaccine against Bacillus anthracis, the causative agent of anthrax, have focused on formulations composed of the protective antigen (PA) adsorbed to aluminum hydroxide. However, due to the labile nature of PA, antigen stability is a primary concern for vaccine development. Thus, there is a need for a delivery system capable of preserving the immunogenicity of PA through all the steps of vaccine fabrication, storage, and administration. In this work, we demonstrate that biodegradable amphiphilic polyanhydride nanoparticles, which have previously been shown to provide controlled antigen delivery, antigen stability, immune modulation, and protection in a single dose against a pathogenic challenge, can stabilize and release functional PA. These nanoparticles demonstrated polymer hydrophobicity-dependent preservation of the biological function of PA upon encapsulation, storage (over extended times and elevated temperatures), and release. Specifically, fabrication of amphiphilic polyanhydride nanoparticles composed of 1,6-bis(p-carboxyphenoxy)hexane and 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane best preserved PA functionality. These studies demonstrate the versatility and superiority of amphiphilic nanoparticles as vaccine delivery vehicles suitable for long-term storage.

Hoping for the best, preparing for the worst: the lived experiences of women undergoing ovarian cancer surgery.

In this study, the lived experiences of women undergoing ovarian cancer surgery were explored, aiming to provide a patient perspective on being newly diagnosed and starting treatment for ovarian cancer. The study period ran from the first visit in the outpatient clinic, till 8 weeks later, when the women had either begun chemotherapy or completed their recovery. Ten women participated in two qualitative research interviews each, before and after surgery. By applying a phenomenological-hermeneutic text interpretation methodology, the findings were systematically identified, put into meaning-structures, interpreted and discussed. This process constituted the theme: 'Hoping for the best, preparing for the worst'. Final diagnostics and treatment start were extensive life events, where life itself was threatened, although hope and will were present. The women intuitively prepared themselves for the diagnosis and treatment. However, the ability to prepare was influenced by personal lifestyle, social conditions, coping strategies, and experiences of hope. The ability to prepare could be strengthened by providing adjusted information, psychosocial support and physical optimisation during the perioperative period. By offering targeted family counselling and taking good care of the women's general health and well-being, hope could be sustained and early cancer rehabilitation initiated.

High-throughput analysis of protein stability in polyanhydride nanoparticles.

Polyanhydrides are a class of surface eroding biomaterials with applications in vaccine and drug delivery. With the complexity and fragile nature of many protein molecules used in therapeutic treatments and vaccines, devices capable of protecting and preserving the functionality of these proteins are essential. In addition, the half-lives of many vaccine antigens and therapeutic proteins are often short, especially at elevated temperatures. In this work a high-throughput methodology has been developed to rapidly assess the effects of polymer chemistry and the various steps during protein delivery (i.e. encapsulation, storage and release) from polyanhydride nanoparticles on the stability of a model protein, bovine serum albumin. Additional factors including microenvironment pH were also investigated in this multi-parametric approach to evaluate protein stabilization. The findings indicate that the microenvironment pH caused by the acidic polymer degradation products was the most detrimental factor affecting protein stability. Nanoparticles based on 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane and 1,6-bis(p-carboxyphenoxy)hexane maintained protein antigenicity over a range of temperatures for 1month. These nanoparticles were also successful in preserving protein structure and emerged as viable candidates for use in future drug/protein delivery applications. The combinatorial approach developed in this work allowed for a 25-fold decrease in time and a 10-fold decrease in the amount of materials needed for the investigation of protein stability when compared to conventional methods.

Novel, high throughput method to study in vitro protein release from polymer nanospheres.

Controlled delivery of therapeutic protein drugs using biodegradable polymer carriers is a desired characteristic that enables effective, application-specific therapy and treatment. Previous studies have focused on protein delivery from polymers using conventional "one-sample-at-a-time" techniques, which are time-consuming and costly. In addition, many therapeutic proteins are in limited supply and are expensive, so it is desirable to reduce sample size for design and development of delivery devices. We have developed a rapid, high throughput technique based on a highly sensitive fluorescence-based assay to detect and quantify protein released from polyanhydrides while utilizing relatively small amounts of protein (approximately 40 microg). These studies focused on the release of a model protein, Texas Red conjugated bovine serum albumin, from polyanhydride copolymers based on sebacic acid (SA) and 1,6-bis(p-carboxyphenoxy)hexane (CPH). The protein release profiles were assessed simultaneously to investigate the effect of polymer device geometry (nanospheres vs films), polymer chemistry, and pH of the release medium. The results indicated that the nanosphere geometry, SA-rich chemistries, and neutral pH release medium led to a more rapid release of the protein compared to the film geometry, CPH-rich chemistries, and acidic pH release medium, respectively. This high throughput fluorescence-based method can be readily extended to study release kinetics for other proteins and polymer systems.

Ultrasound and MRI before and after radical hysterectomy: does the operation affect the pelvic floor muscles?

PURPOSE: To evaluate pelvic floor muscles and bladder neck mobility before and after radical hysterectomy for cervical cancer with magnetic resonance imaging (MRI) and ultrasound. METHODS: A prospective follow-up study of 22 patients studied with MRI and ultrasound preoperatively, three and 12 months after radical hysterectomy was carried out. RESULTS: Bladder neck mobility was decreased three months postoperatively, but one year after the operation, it had almost attained the preoperative level. The size of the pelvic floor muscles was reduced three months after the operation, but one year postoperatively, muscle size had almost returned to the preoperative level. Age and number of deliveries affected the starting point in muscle size and the mobility of the bladder neck, but the development was identical in all the women. CONCLUSIONS: We showed that pelvic floor muscle size and mobility of the bladder neck were reduced three months after radical hysterectomy, and that they regenerated to a level only slightly below the preoperative level one year later.

Differences in current clinical features of diploid and triploid hydatidiform mole.

OBJECTIVE: To describe and compare the current clinical features of diploid and triploid molar pregnancy and to evaluate whether the presenting clinical features can predict the ploidy of a molar pregnancy. DESIGN: A retrospective study of the clinical features and ploidy of hydatidiform moles. SETTING: The Departments of Clinical Genetics and Pathology, Aarhus University Hospital and 13 gynaecological wards, Jutland, Denmark. POPULATION: A total of 259 women with molar pregnancy diagnosed between April 1986 and June 2003. METHODS: A review of medical records of consecutively collected, clinically suspected cases of molar pregnancy was performed. The molar ploidy was determined by karyotyping, flow cytometry, and/or analysis of polymorphic DNA markers. MAIN OUTCOME MEASURES: Maternal characteristics, presenting symptoms, initial human chorionic gonadotrophin (hCG), and molar ploidy. RESULTS: In a multiple logistic regression model, initial hCG of > or = 100,000 iu/l (P < 0.001), first-trimester gestational age (P < 0.001), vaginal bleeding (P < 0.001), and maternal age of > or = 40 years (P = 0.03) were independent predictors of diploid mole. Women with excessive uterine size more frequently had a diploid than a triploid mole (P < 0.001). Fifty-four percent of the women with triploid mole and 27% of the women with diploid mole were diagnosed before onset of symptoms (P < 0.001). CONCLUSIONS: The current clinical features of diploid mole are different from those of triploid mole. The presenting clinical profile of a molar pregnancy may be used as an early predictor of the molar ploidy and thus of the prognosis.

Urogynaecological dysfunction after radical hysterectomy.

AIMS: To identify self-reported urogynaecological dysfunctions in women treated by radical hysterectomy, and predict risk factors. METHODS: A survey using questionnaires. Questionnaires were mailed to 396 patients. RESULTS: Symptoms of urinary incontinence and urinary retention were reported by more than one-third, respectively. One-third of the patients reported cystitis and dysuria. However, 78% of the women did not consider their symptoms as a problem. Analyses identified body mass index, at least one delivery, pre-operative urinary incontinence, and pulmonary disease as predictors for development of post-operative urinary incontinence. Predictive variables for urinary retention symptoms were age, cystitis and/or dysuria, previous rupture of the anal sphincter, fetal weight at delivery > or =4000 g, sensation of vaginal dryness, and pre-operative urinary retention symptoms. Considering post-operative cystitis and/or dysuria, fetal weight > or =4000 g, sensation of vaginal dryness, cystitis and/or dysuria before the operation, and physical activity were predictive variables. CONCLUSIONS: Patients can be informed and advised about possible urinary tract symptoms. Special attention in the pre- and post-operative period can be paid to a subgroup of patients at high risk of later urogynaecological problems.

Patients with ovarian cancer have elevated (51)Cr-EDTA plasma clearance early post-operatively.

Plasma clearance of (51)Cr-EDTA (Clp(EDTA)) is widely used to determine glomerular filtration rate prior to carboplatin based chemotherapy. We have observed that many patients with ovarian cancer have elevated Clp in the early post-operative phase compared to later phases. The purpose of this study was to examine whether this observation reflects a systematic difference. We retrospectively analysed data from 53 patients who had undergone surgery for ovarian cancer. Twenty-six patients had Clp(EDTA) measured early after the operation (mean, 8 days (range, 3-16 days)) (early group), and 27 patients had Clp(EDTA) measured late post-operatively (mean, 32 days (range, 19-48 days)) (late group). Clp(EDTA) values was measured before the first, third and fifth course of chemotherapy. Additionally, age, height, weight, cancer stage, ascites and tumour histology were noted. Mean Clp(EDTA) in the early group was significantly higher than in the late group (104+/-4.4 vs 89+/-3.5 ml.min(-1) per 1.73 m(2); P =0.005). Clp(EDTA) declined significantly in the early group from the first measurement after the operation until measurement before the third course of chemotherapy but remained constant in the late group. Clp(EDTA) was not correlated to ascites, cancer stage or tumour histology. It is concluded that patients with ovarian cancer have significantly higher Clp(EDTA) in the early post-operative phase than similar patients with Clp(EDTA) measured late post-operatively.

No effects of human relaxin on the active and passive biomechanical properties of isolated cervical specimens from nonpregnant women.

OBJECTIVE: To evaluate the effect of human relaxin (hRLX-2) on the active and passive biomechanical properties of cervical tissue in vitro. MATERIAL: Cervical samples were obtained from the middle part of the cervix in 22 nonpregnant women undergoing hysterectomy. METHODS: The effect of hRLX-2 (10(-7) M) on the active biomechanical properties was studied on vasopressin (10(-8) M) induced smooth muscle contractions in an organ bath model. The effect on the passive biomechanical properties were studied after incubation of the strips for 48 h with hRLX-2 (10(-8) M and 10(-9) M). Subsequently, the specimens were stretched in a material testing machine until they broke. The load applied and the elongation were simultaneously recorded and the results translated into stress-strain curves. RESULTS: hRLX-2 did not influence the vasopressin-induced contractility of cervical strips from nonpregnant women in this study. No synergistic effect of progesterone could be demonstrated. The passive biomechanical properties (tensile strength, extensibility, stiffness of failure energy) did not change significantly after relaxin incubation. The results obtained in vitro do not suggest an important physiological effect of relaxin on the human nonpregnant cervix.

Cervical collagen in non-pregnant women with previous cervical incompetence.

OBJECTIVE: To study the cervical collagen in non-pregnant women with a history of congenital cervical incompetence. SUBJECTS: Ten non-pregnant women with congenital cervical incompetence, defined as cervical incompetence in the first pregnancy in a woman without previous cervical trauma. Seventy-one normal non-pregnant women (31 nulligravidae, nine nulliparous and 31 parous women) served as controls. METHOD: Cervical biopsies were analysed for the hydroxyproline concentration and extractability to express the concentration and the stability of the collagen. RESULTS: Women with cervical incompetence had markedly lower median cervical hydroxyproline concentration when compared to normal parous women (11.1 (range 6.11-18.5) micrograms/mg wet weight vs. 16.9 (range 7.7-29.8) micrograms/mg wet weight; P = 0.003, Mann Whitney test) and the extractability was almost twice the value found in normal parous women (80.2% vs. 49.5%; P = 0.03, Mann Whitney test). Both pregnancy per ce and parturition caused a significant decrease in the hydroxyproline concentration when compared to values found in nulligravidae (P < 0.0001, Kruskall Wallis test). CONCLUSIONS: Congenital cervical incompetence seems associated with a low collagen concentration in the cervical connective tissue in the non-pregnant state.

Unmeasurable relaxin concentrations in repeated pregnancies after embryo transfer.

Unmeasurable serum relaxin concentrations were found in repeated serum samples taken during two subsequent pregnancies in a woman pregnant after in vitro fertilization in artificial cycles. Despite the absence of relaxin both pregnancies were carried out normally until term except for symptoms of pelvic girdle relaxation. Labour induction was attempted at term as placental insufficiency was suspected. However, the cervical score remained unfavourable and Cesarian section was performed. In the second pregnancy elective Cesarian section was performed due to mild preeclampsia. This case indicates that relaxin may not be important for either suppression of myometrial activity or pelvic girdle relaxation during human pregnancy.

Relaxin is not related to symptom-giving pelvic girdle relaxation in pregnant women.

BACKGROUND: The pregnancy associated hormone relaxin induces loosening of the pelvic ligaments in several species. This study was undertaken to evaluate whether pregnant women with symptom-giving girdle relaxation had increased serum relaxin concentrations during pregnancy. METHOD: Serum relaxin concentrations were measured in 38 pregnant women with symptom-giving pelvic girdle relaxation at the time of diagnosis, in the 30th and 38th week of pregnancy as well as 2 and 6 months after delivery. Fourteen pregnant women without symptoms served as a control group. Relaxin concentrations were measured by a homologous enzyme linked immuno-sorbent assay. All participants were clinically examined including tests for symptom-giving pelvic girdle relaxation. RESULTS: No differences in serum relaxin concentrations were found throughout pregnancy and after delivery. CONCLUSION: The present results do not suggest an important role for relaxin in symptom-giving pelvic girdle relaxation during human pregnancy.

Variations in serum relaxin (hRLX-2) concentrations during human pregnancy.

STUDY OBJECTIVE: To study variations in serum relaxin concentrations during normal and abnormal human pregnancy and parturition and in umbilical cord blood. DESIGN AND PATIENTS: Diurnal variations were determined in samples collected every 2 hours for 24 hours (n = 4). Variations during pregnancy were studied in samples taken every 4th week from normal pregnant women (n = 26). Additionally samples were collected once a week from the 37th week of pregnancy and until spontaneously delivery (n = 5). Changes in relaxin during early pregnancy (gestational age: 30 to 97 days) were studied in serum from 12 normal pregnant women, 13 with spontaneous abortion and 38 with an ectopic pregnancy. Fetal serum was obtained at delivery from the umbilical vein (n = 20). All samples were analysed for relaxin by an ELISA based on human relaxin antibodies. MAIN RESULTS: No diurnal variation in relaxin concentrations were found. In normal pregnant women relaxin concentrations increased during the first 14 weeks of pregnancy. From week 14 to 24 a gradual decrease was found and the concentrations remained constant during the last part of pregnancy. No changes in concentrations were found immediately before or during spontaneous delivery. Women with abnormal pregnancies had lower serum relaxin values than normal early pregnant women. The relaxin concentration was significantly correlated to the serum HCG concentration in early pregnancy but not to the serum HPL levels after week 28. In the umbilical vein relaxin concentrations were low. CONCLUSION: The variation in circulating levels of relaxin during human pregnancy differs markedly from those found in other species. This suggests a different role for relaxin in human pregnancy.

Morphological, stereological, and biochemical analysis of the mini-pig urinary bladder after chronic outflow obstruction and after recovery from obstruction.

Chronic partial bladder outlet obstruction was created in nine mini-pigs by implanting a 6-7 mm ring around the proximal urethra. After a median obstruction period of 63 days, the ring was removed and after a recovery period of median 60 days the animals were sacrificed. Changes in muscle and connective tissue were assessed by unbiased, modern morphometry and biochemical analysis. After obstruction the results were as follows: (1) a 6-fold increase in bladder weight, (2) a 2.5-fold increase in smooth muscle cell size, (3) a 3-fold increase in smooth muscle cell number, (4) unchanged proportions between muscle and connective tissue, (5) unchanged hydroxyproline concentrations, (6) an 8-fold increase in total collagen content, (7) an increase in the ratio of type I/III collagen, and (8) a 7-8-fold increase in total content of type I and III collagen. All changes were markedly, though incompletely, reversed after recovery, except smooth muscle cell number and the ratio of type I/III collagen.

Relaxin (hRLX-2)-induced weakening of human fetal membranes in vitro.

We examined the effect of human relaxin (hRLX-2) on the biomechanical properties of human fetal membranes in vitro. Intact chorioamniotic membranes were obtained from twelve elective cesarean sections before the onset of labor. Membrane strips with a fixed width were biomechanically tested after incubation for 20 h with hRLX-2 in concentrations of 10(-7) mol/l and 10(-9) mol/l. Incubation with hRLX-2 (10(-9) mol/l) changed the load-strain values as the membrane stiffness was decreased by 19% when compared with controls: median 2.45 N (range, 0.81-4.31) versus 3.03 N (1.28-5.46), P = 0.02 (Mann-Whitney test). For description of the membrane material as such, the stress-strain values were calculated by dividing the load-strain values with the cross sectional area of the membranes. Incubation with hRLX-2 (10(-9) mol/l) decreased the tensile strength of the membranes by 30%-0.817 N/mm2 (0.282-1.139) vs. 0.575 N/mm2 (0.101-1.150), P = 0.03--and reduced the elastic modulus by 31%--2.26 N/mm2 (0.82-5.08) versus 1.57 N/mm2 (0.51-3.71), P = 0.002. Less pronounced effects were found after incubation with hRLX (10(-7) mol/l). No quantitative or qualitative changes of the membrane collagen were found after relaxin incubation. Although the mechanism for rupture of the fetal membranes remains unknown, the present results suggest that relaxin might be involved in the process leading to rupture of the membranes.

Normal serum relaxin in women with disabling pelvic pain during pregnancy.

As part of a case control within cohort study 472 pregnant women answered a questionnaire post partum. Serum relaxin concentrations were measured by a homologous ELISA in samples collected from the women in the 30th week of pregnancy. Serum relaxin concentrations were not associated with pregnancy-associated pelvic pain. Thus, normal pregnant women without pelvic pain (n = 118) had mean concentrations of 343 pg/ml compared to 332 pg/ml in women (n = 59) with pelvic pain and some restriction in daily activities and to 349 pg/ml in women (n = 9) with severe pregnancy-induced pelvic pain. The present results do not suggest a role for relaxin in symptom-giving pelvic relaxation during human pregnancy.