Rapid screening of infertility-associated gynecological conditions via ambient glow discharge mass spectrometry utilizing urine metabolic fingerprints.

Infertility presents a widespread challenge for many families worldwide, often arising from various gynecological diseases (GDs) that hinder successful pregnancies. Current diagnostic methods for GDs have disadvantages such as low efficiency, high cost, misdiagnose, invasive injury and etc. This paper introduces a rapid, non-invasive, efficient, and straightforward analytical method that utilizes desorption, separation, and ionization mass spectrometry (DSI-MS) platform in conjunction with machine learning (ML) to detect urine metabolite fingerprints in patients with different GDs. We analyzed 257 samples from patients diagnosed with polycystic ovary syndrome (PCOS), premature ovarian insufficiency (POI), diminished ovarian reserve (DOR), endometriosis (EMS), recurrent pregnancy loss (RPL), recurrent implantation failure (RIF), and 87 samples from healthy control (HC) individuals. We identified metabolite differences and dysregulated pathways through dimensionality reduction methods, with the result of the discovery of 7 potential biomarkers for GDs diagnosis. The ML method effectively distinguished subtle differences in urine metabolite fingerprints. We anticipate that this innovative approach will offer a patient-friendly, rapid screening, and differentiation method for infertility-related GDs patients.

Population based study of genital Chlamydia trachomatis prevalence and associated factors in Norway: a cross sectional study.

BACKGROUND: The number of diagnosed cases of Chlamydia trachomatis infection has been increasing in the past years in Norway although the testing rate has been relatively stable. The aim of this study was to measure the prevalence of genital Chlamydia trachomatis in young men and women in one county in Norway and determine associated factors in order to better target preventive measures. METHODS: We mailed to a random sample of 10,000 persons aged 18-25 in Rogaland county a mail-back urine sample kit and a self-administered questionnaire with questions on socio-demographic details, health seeking behaviour and symptoms of and history of sexually transmitted diseases. Associations between current Clamydia trachomatis infection and the above mentioned factors were studied by multiple logistic regression. RESULTS: The response rate among women was 18.9% (930/4923) and 11.9% (605/5077) among men. The prevalence of Chlamydia trachomatis infection was 5.8% (95% CI 4.5-6.8) among women and 5.1% (95% CI 3.8-6.8) among men. For men a greater number of partners during the last year (p for trend < 0.001), and living in a municipality without a local youth clinic increased the odds of infection (OR 8.6, 95% CI 2.2-33.9). For women a greater number of partners during the last year (p < 0.001) and not having consulted a family doctor for STIs (OR 2.1 95% CI 1.1-4.2) were positively associated with infection while not having a previous Chlamydia trachomatis diagnosis decreased the odds of having this infection (OR 0.3, 95% CI 0.2-0.7). CONCLUSION: Our results indicate the importance of having a visible youth clinic in each municipality. It also suggests targeting women who have had a previous Chlamydia trachomatis infection diagnosed before.

Associations of exposure to phthalates and environmental phenols with gynecological disorders.

Phthalates and environmental phenols might be associated with some benign diseases that have been found to be hormone-sensitive. Current knowledge on adverse effects of these chemicals among reproductive women is limited and often controversial. Therefore, the purpose of this study was to investigate the association between the urinary concentration of phthalates and environmental phenols and gynecological disorders from 512 women of reproductive age. The association between chemical concentration and disease in the control and case groups was statistically determined with the questionnaire survey data and measurements using the LC-MS/MS. The results have shown that DEHP metabolites, ethyl paraben and 3,4-DHB showed significant direct associations with leiomyoma and benign ovarian tumors (p < 0.05). We found statistically significant positive relationships between exposure to chemicals (some DEHP metabolites, DHB) and prevalence of gynecologic disorders (p < 0.05). Furthermore, the ORs for leiomyoma associated with these compounds in always user for personal care products (PCPs) was higher than those of sometimes user. High levels of urinary concentrations of these compounds such as DEHP metabolites and parabens and their metabolites showed significant associations with leiomyoma and benign ovarian tumors.

The purification and development of a radioimmunoassay for beta-core fragment of human chorionic gonadotrophin in urine: application as a marker of gynaecological cancer in premenopausal and postmenopausal women.

The beta-core fragment of human chorionic gonadotrophin (hCG) is a major part of the immunoreactive hCG-like material found in the urine of normal pregnant women. Patients with non-trophoblastic gynaecological malignancies have been found to have raised levels of urinary beta-core. We describe the purification of beta-core, the preparation of a polyclonal sheep antiserum and the development of radioimmunoassay. The minimum detection limit of this assay was 0.025 micrograms beta-core/l. There was no significant cross-reaction with the free alpha-subunit, hLH, hFSH and hTSH (less than 0.7%), and only partial cross-reaction with intact hCG and free beta-subunit of hCG (6.9 and 18%). Within-assay variability ranged from 2.03 to 12.5% and between-assay variability from 2.25 to 13.4%. The assay was applied to urine samples from 92 normal non-pregnant premenopausal women, 54 normal postmenopausal women and 65 women with active gynaecological disease (47 postmenopausal and 18 premenopausal). In normal premenopausal women the values ranged from less than 0.025 to 0.62 micrograms beta-core/l (median 0.043 micrograms beta-core/l). The values for normal postmenopausal women ranged from less than 0.025 to 0.64 micrograms beta-core/l (median 0.26 micrograms beta-core/l). Postmenopausal women with gynaecological malignancy had values which ranged from less than 0.025 to 4.0 micrograms/beta-core/l (median 0.31 micrograms beta-core/l); premenopausal women in this group had values which ranged from less than 0.025 to 1.15 micrograms beta-core/l (median 0.12 micrograms beta-core/l). On molecular sieve chromatography, the material found in the urine of normal postmenopausal women showed the physicochemical characteristics of authentic beta-core.(ABSTRACT TRUNCATED AT 250 WORDS)

Usefulness of microscopic examination in urinalysis.

The authors evaluated 500 patients who yielded urine specimens containing red and white blood cells but that gave negative reagent test strip reactions to determine the medical usefulness of the microscopic examination in such cases. Half of the patients had a history of, or current signs and symptoms of genitourinary or renal disease (GURD), or had indwelling catheters, hypertension, or diabetes. The other half did not display these conditions. Red blood cells occurred rarely, and no red blood cell associated GURD was detected in these patients. Five had lower urinary tract infection. Seventy-two underwent further workup, but no GURD was found. Physicians did not comment or take action on the report in other patients. The authors found the test for leukocyte esterase and nitrite (LN) to have a predictive value for a negative result of 97% for exclusion of bacteruria. Based on these observations, the authors established in 1982 a policy that microscopic examination would be performed only on specimens negative by reagent test strip (including LN) if a "diagnostic urinalysis" (DU) was ordered. The authors recommended that DU be requested only for patients suspected of GURD. This has eliminated microscopic examinations on 25% of specimens and reduced costs.

Dipstick testing of urine--can it replace urine microscopy?

One thousand consecutive urine specimens were studied to assess the sensitivity of a commercially available dipstick (Chemstrip 8, Boehringer Mannheim Corp., Indianapolis, IN) to predict the presence or absence of microscopic abnormalities. The Chemstrip 8 had a sensitivity of 78%, specificity of 54%, and a false negative rate of 38%. An additional 1,000 consecutive urine specimens were then studied using the Chemstrip 9, a reagent dipstick that includes the leukocyte esterase (LE) test. The Chemstrip 9 had a sensitivity of 82%, specificity of 42%, and a false negative rate of 36%. Chi-squared analysis revealed that the two dipsticks were not significantly different (chi 2 = 0.17, P greater than 0.5). Clinical review of patients with false negative results showed that approximately one-third to one-half of these patients had either spinal cord injury or genitourinary problems. Maximal potential savings in workload of approximately 10% were found if microscopic examinations were to be performed only on urine specimens with abnormal dipsticks. Our data suggest that in our patient population, we should not eliminate microscopic urine examination based on abnormal dipstick findings.

Schistosomiasis of the lower reproductive tract without egg excretion in urine.

The individual and public health impact of female genital schistosomiasis (FGS) has been studied and FGS as a risk factor for acquiring human immunodeficiency virus is discussed. In a community-based study in Tanzania, 40% of the women of child-bearing age (n=543) showed excretion of Schistosoma haematobium eggs in the urine (median=2.2 eggs/10 ml of urine) and 32% (n=263) had S. haematobium eggs in their cervical tissue. Urinary and genital schistosomiasis coexisted in 62% of the women, but S. haematobium eggs were found in the cervix without detectable egg excretion in the urine in 23%. Only 43% of the FGS cases had hematuria. Since FGS frequently exists in women with scanty or no egg excretion in the urine and because this disease manifestation is a considerable individual and public health hazard in S. haematobium-endemic areas, mass treatment targeted to women of child-bearing age should be considered.

Urine reagent strips for diagnosis of schistosomiasis haematobium in women of fertile age.

Hematuria, proteinuria and leukocyturia were semiquantitatively assessed by reagent strips in single morning urine of women of fertile age visiting the outpatient department of the Mangochi district hospital, Malawi. This was part of a diagnostic approach to female genital schistosomiasis (FGS). In 51 women ova of Schistosoma haematobium were detected in urine by a filtration technique. In 33 of these women ova were also present in genital tissue as demonstrated by microscopic examination of biopsies. In 209 women no ova were found in the single urine filtered. There were significantly higher scores for hematuria, proteinuria and leukocyturia as well as of the combined reagent strip index (RSI) in egg-excreting than in egg-negative women. The sensitivity of a single hematuria, proteinuria and leukocyturia reading was 98, 84 and 73%, respectively. However, the respective specificity was only 24, 22 and 23%. The best prediction of urinary schistosomiasis was achieved by a +2 score for hematuria, of which the sensitivity was 94% and the specificity was 61%. The high false-positive rates can probably be explained by contamination of urine by vaginal secretion. Moreover, cases of schistosomiasis have probably been overlooked because only a single morning urine sample was examined. The total absence of hematuria, proteinuria and leukocyturia, however, may be used to rule out heavy infections in community surveys. There was no difference in reagent strip scores between women with genital and urinary schistosomiasis as compared with those with urinary tract lesions alone. Thus urine analysis reagent strip readings do not help to discriminate between S. haematobium infected women with and without FGS.

Female genital schistosomiasis due to Schistosoma haematobium. Clinical and parasitological findings in women in rural Malawi.

A total of 51 women with urinary schistosomiasis haematobium were examined in order to identify diagnostic indicators for female genital schistosomiasis (FGS). Patients were selected at random from the outpatient department of the Mangochi District Hospital, Malawi. The medical histories were recorded according to a pre-designed questionnaire and the women were subjected to a thorough gynaecological examination including colposcopy and photographic documentation of lesions. Microscopy of genital biopsies revealed that 33 of the 51 women had S. haematobium ova in cervix, vagina and/or vulva in addition to the presence of ova in urine. The most sensitive diagnostic procedure was beside microscopic examination of a wet cervix biopsy crushed between two glass slides, which revealed 25 of the 33 genital infections. There was a significant correlation between the size of genital lesions and the number of ova counted per mm2 of crushed tissue. Women with FGS had significantly more tumours in the vulva than women with schistosomiasis limited to the urinary tract. Most of the observed genital pathology could easily be identified by the naked eye, but colposcopic examination yielded valuable additional information like the demonstration of neovascularisation around cervical sandy patches. Few of the symptoms previously regarded as indicators for FGS could be linked to the presence of schistosome ova in genital tissue. Husbands of infertile women with FGS had children with other women significantly more often than husbands of women who only had urinary schistosomiasis. This, together with the finding that the majority of the divorced women had FGS, indicates that the manifestation of this disease may have implications for the marital and sexual life of the affected women.

Reversibility of lower reproductive tract abnormalities in women with Schistosoma haematobium infection after treatment with praziquantel--an interim report.

Little is known whether and to what extent antiparasitic treatment cures female genital schistosomiasis (FGS). Using a standard protocol, of twenty-one women with FGS nine were re-examined at two to nine weeks after they had been treated with praziquantel at a single dose of 40 mg/kg. Symptoms related to pathology of the urinary tract and to a lesser extent of genital pathology subsided in most patients. Schistosoma haematobium ova were no longer detectable in urine of any of the patients post-treatment. Efficiency of chemotherapy against adult worms was confirmed by the disappearance of circulating anodic antigen (CAA) in serum. Sandy patches showed resolution in two of four cases after chemotherapy. Papillomata due to schistosomiasis alone improved, but persisted in mixed infection with human papilloma virus (HPV) or when HPV was the only underlying cause. In one patient ulcera could not be related with certainty to schistosomiasis at admission, but resolved after treatment with parziquantel. Leukoplakia (two cases) was not influenced by chemotherapy, or even increased during follow-up, regardless of whether ova had been detected or not. Although the follow-up period was rather short, time intervals were not standardized, and a relatively small number of patients was investigated, it could be shown that genital pathology due to sequestered S. haematobium ova is, at least partially, reversible already two to nine weeks after killing the adult worms by praziquantel. This is paralleled by a normalization of inflammatory immune responses detectable in histological sections and vaginal lavage.

Combined use of urinary UGP and serum CA 125 in the diagnosis of gynecological cancers.

UGP, the beta-core fragment of human chorionic gonadotropin has been proposed as a tumor marker for gynecological malignancies. This fragment may be detected in a single morning-specimen of urine using an enzyme immunoassay. In this study, the diagnostic usefulness of urine UGP and serum CA 125 measurement for gynecological neoplasias (149 cases) was evaluated using a control group of patients with benign gynecological diseases (69 cases) and healthy females (99 cases). Considering the neoplastic patients in comparison to patients with benign diseases, the best diagnostic efficiency (78%) was found to correspond to a cut-off level of 120 pmol/mol creatinine the sensitivity being 73% and the specificity 90%. With this cut-off, an efficiency of 82% for healthy controls was obtained. Since the menopausal condition increases UGP levels, and though no significant difference for UGP was found between healthy subjects and patients with benign diseases, we decided to consider the reference populations as a single group. Thus, we evaluated the UGP performance on the basis of menopausal status. When a specificity of 95% was fixed, the cut-off values were 120 and 180 pmol/mol creatinine for pre- and postmenopausal women respectively, the sensitivity being 73% and 64%. Finally the combined evaluation of UGP and CA 125 improved their individual clinical efficiency for the diagnosis of ovarian serous cystadenocarcinomas, assuring a sensitivity of 86% and a specificity of 89%.

Urinary gonadotropin peptide (UGP) as a marker of gynecologic malignancies.

Urinary gonadotropin peptide (UGP) was measured in 866 urines from normal women and women with benign and malignant gynecologic disease using the Triton UGP enzyme immunoassay. The greatest level of overexpression of the marker was observed in patients with ovarian cancer. Using a cutoff of 4 fmol/mg creatinine, UGP was overexpressed in samples from 2% of normal premenopausal women, 15% of normal postmenopausal women, 5% of women with benign gynecologic disease, and 59% of women with ovarian cancer. UGP expression was independent of the histologic type of ovarian cancer. The expression of UGP and CA 125 were not correlated and use of the two markers in tandem increased the sensitivity of detection of disease by greater than 20% over that which was observed using each marker individually. UGP levels were correlated with clinical status, and doubled in value in 67% of patients with progressive disease, and were halved in 93% of patients who were in remission at the time of the study.

[Preliminary experience with a new tumor marker in obstetrics and gynecology: UGP (Urinary Gonadotropin Protein)]. / Esperienze preliminari con un nuovo marcatore tumorale in ostetricia e ginecologia: UGP.

AIM: To evaluate the use of UGP (urinary gonadotropin protein) as a tumor marker in gynaecologic and obstetric malignant diseases. MATERIALS AND METHODS: The study was carried out in the division of Gynaecology and Obstetrics of the Ospedali Riuniti in Bergamo. 63 patients, with obstetric or gynaecologic benign or malignant diseases, entered the study. 66 healthy volunteers were examined as a group-control. In both the groups UGP levels were determined in morning urine, using an immunoenzymatic commercial kit. RESULTS: Results, expressed in fmol UGP/ml of urin, show that UGP is produced by several neoplasms, but the false-positive percentage is still high; a higher precision can be obtained with an accurate choice of the cut-off value and with a standardization of the analytical technics. Besides, the contemporary determination of UGP and CA 125 levels reduces the possibility of false-positive and false-negative results. CONCLUSIONS: More studies must be carried out to confirm the value of UGP as a tumor marker in obstetrics and gynaecology. Anyway, this recently purified protein can already be useful, in combination with the usual tumor markers, in the prompt diagnosis and management of primary neoplasms or recurrences, with a higher sensibility in comparison with traditional clinical and radiological examinations.

[Systematic home screening for Chlamydia trachomatis infections of asymptomatic men and women in family practice by means of mail-in urine samples]. / Systematische opsporing van infecties met Chlamydia trachomatis bij mannen en vrouwen zonder klachten in de huisartspraktijk met behulp van per post verstuurde urinemonsters.

OBJECTIVE: To determine the prevalence and determinants of Chlamydia trachomatis (CT) infections among asymptomatic men and women in general practice. To determine participation rates in a systematic screening programme in general practice, using home obtained mailed urine samples. DESIGN: Cross-sectional study. METHODS: In 15 general practices in Amsterdam, the Netherlands, a sample of 11,005 persons (5541 women and 5464 men), aged 15-40 were invited to send in a urine sample and a completed questionnaire by mail. The urine samples were tested using the ligase chain reaction for DNA amplification. Patients diagnosed with CT were treated and partner notification was performed. RESULTS: 33% of invited males (1809/5464) and 50% of females (2751/5541) sent in the study material. Older patients participated more frequently than younger patients. Participation rates among persons with a Dutch background were higher than rates among persons from other ethnic groups. In 42 men and 79 women a CT infection was identified (2.3% and 2.9% respectively). Infections were more prevalent in patients from Surinam and the Dutch Antilles and in the age category 21-25 years. Type of health insurance as a proxy measure of socioeconomic status was not an indicator of infection. CONCLUSION: The participation in this systematic screening using mail-sent urine samples was 33% in men and 50% in women. The CT prevalences among asymptomatic men and women were 2.3% and 2.9% respectively.

Practical uses of steroids and gonadotropins in obstetrics and gynecology.

PIP: Several steroid and gonadotropin determinations that are available to practicing physicians are discussed and their practical uses are emphasized. Before undertaking a workup of a patient who appears to have an endocrine abnormality a physician should know the availability and benefits of such tests, the time and expense involved, and the interpretation of laboratory reports. Estrogens are phenolic steroids secreted by the ovaries, adrenal glands, testicles, and the fetal-placental unit and are found in over 20 metabolites in the urine; the most important are estrone, estridiol, and estriol. Total or fractional determinations of urinary estrogens are expensive and may require 7 days. Human gonadotropin determinations are expensive and require 2 weeks. Contraceptive medications, tranquilizers, and sedatives interfere with secretion of this hormone. Clinical usefulness is of value only when dterminations are extremely high or repeatedly ver y low. Culdoscopic findings may be of more value than either estrogen or gonadotrpin determination. Estrogen determinations can be of value in male patients with gynecomastia, in suspected fetal distress or fetal death in utero (a drop of 50% in the normal 1000-fold increase of pregnancy indicates need for delivery of infant), and when an indication of size of infant is needed to determine elective repeat ceasarean sectton. Ther is no practical value of pregnanediol determination in pregnancy or in the amenorrheic female. The clinical use of 17-hydroxycorticoids is related to its value as a screening procedure for adrenal disorders. Patient stress and several drugs may interfere with accurate tests. The 17-ketosteroids are used primarily for the sam e purpose. Progesterone determinations are useful to differentiate ovarian from adrenal pathology. Urinary testosterone increase may be present in some ovarian tumors. Human chorionic gonadotropin (HCG) detection is used for pregnancy tests; a sensitive technique may obtain a positive test day Day 30 of the menstrual cycle or 16 days after conception. These tests are also used in the diagnosis of hydatidiform mole, choriocarcinoma, and to guide care of trophoblastic disease. Of newer methods the competitive protein binding technique for estrogen, progesterone, and testosterone is rapid, accurate, and sensitive. Radioimmunoassay for FSH, LH, and other trophic hormones is complex but promising. Conversion studies are complex but may be of use.^ieng

[LH-excretion in healthy omen and in patients with gynecologic-endocrine diseases]. / A luteinizáló hormon (LH) ürítés vizsgálata egészségeseken és nögyógyászati endocrin betegségekben

PIP: Within the framework of gynecologic-endocrine examinations, the authors tested with an immunochemical method using Luteonosticon, the excretion of luteinizing hormone in 4 healthy women and in 18 patients with endocrinal disorders. The method gave modest results in the case of primary amenorrhea; however in the case of secondary amenorrhea, the excretion of luteinizing hormone proved to be very useful in establishing a diagnosis, particularly when the absence of bleeding is accompanied by infertility. The method was found to be simple and very useful in establishing diagnosis in gynecologic-endocrine disorders.^ieng

A simplified method to recover urinary vesicles for clinical applications, and sample banking.

Urinary extracellular vesicles provide a novel source for valuable biomarkers for kidney and urogenital diseases: Current isolation protocols include laborious, sequential centrifugation steps which hampers their widespread research and clinical use. Furthermore, large individual urine sample volumes or sizable target cohorts are to be processed (e.g. for biobanking), the storage capacity is an additional problem. Thus, alternative methods are necessary to overcome such limitations. We have developed a practical vesicle isolation technique to yield easily manageable sample volumes in an exceptionally cost efficient way to facilitate their full utilization in less privileged environments and maximize the benefit of biobanking. Urinary vesicles were isolated by hydrostatic dialysis with minimal interference of soluble proteins or vesicle loss. Large volumes of urine were concentrated up to 1/100 of original volume and the dialysis step allowed equalization of urine physico-chemical characteristics. Vesicle fractions were found suitable to any applications, including RNA analysis. In the yield, our hydrostatic filtration dialysis system outperforms the conventional ultracentrifugation-based methods and the labour intensive and potentially hazardous step of ultracentrifugations are eliminated. Likewise, the need for trained laboratory personnel and heavy initial investment is avoided. Thus, our method qualifies as a method for laboratories working with urinary vesicles and biobanking.

Detection of the HE4 protein in urine as a biomarker for ovarian neoplasms.

The HE4 protein is overexpressed in ovarian carcinomas and can be detected in serum by an ELISA with sensitivity similar to CA125 and higher specificity for malignant disease. We now demonstrate that HE4 can also be detected in the urine at a specificity level of 94.4%, including 13/15 (86.6%) with stage I/II and 57/64 (89.0%) with stage III/IV disease and including 90.5% of patients with serous ovarian carcinoma. Assaying serum and urine from the same patients showed similar sensitivity. Our data indicate that measuring HE4 in urine may aid diagnosis and the monitoring of response to therapy.