Multiplexed micronutrient, inflammation, and malarial antigenemia assessment using a plasma fractionation device.

Processing and storing blood samples for future analysis of biomarkers can be challenging in resource limited environments. The preparation of dried blood spots (DBS) from finger-stick collection of whole blood is a widely used and established method as DBS are biosafe, and allow simpler field processing, storage, and transport protocols than serum or plasma. Therefore, DBS are commonly used in population surveys to assess infectious disease and/or micronutrient status. Recently, we reported that DBS can be used with the Q-plex™ Human Micronutrient 7-plex Array (MN 7-plex), a multiplexed immunoassay. This tool can simultaneously quantify seven protein biomarkers related to micronutrient deficiencies (iodine, iron and vitamin A), inflammation, and malarial antigenemia using plasma or serum. Serum ferritin, an iron biomarker, cannot be measured from DBS due to red blood cell (RBC) ferritin content confounding the results. In this study, we assess a simple blood fractionation tool that passively separates plasma from other blood components via diffusion through a membrane into a plasma collection disc (PCD). We evaluated the concordance of MN 7-plex analyte concentrations from matched panels of eighty-eight samples of PCD, DBS, and wet plasma prepared from anticoagulated venous whole blood. The results showed good correlations of >0.93 between the eluates from PCD and DBS for each analyte except ferritin; while correlations seen for plasma/PCD were weaker. However, the recovery rate of the analytes from the PCD were better than those from DBS. The serum ferritin measures from the PCD were highly correlated to wet plasma samples (0.85). This suggests that surveillance for iron status in low resource settings can be improved over the current methods restricted to only measuring sTfR in DBS. When used in combination with the MN 7-plex, all seven biomarkers can be simultaneously measured using eluates from the PCDs.

A case series to examine the perinatal outcomes of infants conceived by intravaginal culture (IVC).

PURPOSE: In vitro fertilization (IVF) has been a well-established method for treating infertility for over four decades. The mainstay method of culture of oocytes and embryos has been in gas incubators. More recently, the novel use of a gas-permeable closed vessel to culture oocytes and embryos in the vagina, intravaginal culture (IVC), has been introduced as a viable lower-cost option for infertility patients. Several studies have studied the efficacy of IVC; however, there is no data on the perinatal outcomes of the babies born using this newer technology. METHODS: Our study is a retrospective case series (n = 66) from a single center, uniquely examining the perinatal outcomes of infants born after IVC. RESULTS: There were 50 singleton and 16 twin gestations in this case series. For singleton infants conceived via IVC (n = 50), the mean gestational age at delivery was 38 weeks and 4 days, and the mean birth weight was 3159.1 + / - 501.5 g. Four infants were born with low birth weight, three were born preterm, and one was born macrosomic. The twin pregnancies had a mean gestational age at delivery of 33 weeks 4 days and a mean birth weight of 1992.9 + / - 620.7 g. Twenty-seven infants met the criteria for low birthweight, and twenty-four infants delivered preterm. No twin infants met the criteria for macrosomia. CONCLUSION: This case series provides an initial description of the perinatal outcomes of IVC conceived infants, which shows no concerning trends in adverse birth outcomes for singleton infants. As expected, IVC twin gestations had a high rate of low birth weight and preterm delivery. Continued larger studies are essential to provide more comprehensive data on perinatal outcomes of infants conceived by this new technology.

A multicenter analytical performance evaluation of a multiplexed immunoarray for the simultaneous measurement of biomarkers of micronutrient deficiency, inflammation and malarial antigenemia.

A lack of comparative data across laboratories is often a barrier to the uptake and adoption of new technologies. Furthermore, data generated by different immunoassay methods may be incomparable due to a lack of harmonization. In this multicenter study, we describe validation experiments conducted in a single lab and cross-lab comparisons of assay results to assess the performance characteristics of the Q-plex™ 7-plex Human Micronutrient Array (7-plex), an immunoassay that simultaneously quantifies seven biomarkers associated with micronutrient (MN) deficiencies, inflammation and malarial antigenemia using plasma or serum; alpha-1-acid glycoprotein, C-reactive protein, ferritin, histidine-rich protein 2, retinol binding protein 4, soluble transferrin receptor, and thyroglobulin. Validations included repeated testing (n = 20 separately prepared experiments on 10 assay plates) in a single lab to assess precision and linearity. Seven independent laboratories tested 76 identical heparin plasma samples collected from a cohort of pregnant women in Niger using the same 7-plex assay to assess differences in results across laboratories. In the analytical validation experiments, intra- and inter-assay coefficients of variation were acceptable at <6% and <15% respectively and assay linearity was 96% to 99% with the exception of ferritin, which had marginal performance in some tests. Cross-laboratory comparisons showed generally good agreement between laboratories in all analyte results for the panel of 76 plasma specimens, with Lin's concordance correlation coefficient values averaging ≥0.8 for all analytes. Excluding plates that would fail routine quality control (QC) standards, the inter-assay variation was acceptable for all analytes except sTfR, which had an average inter-assay coefficient of variation of ≥20%. This initial cross-laboratory study demonstrates that the 7-plex test protocol can be implemented by users with some experience in immunoassay methods, but familiarity with the multiplexed protocol was not essential.

Night shift work, chronotype and prostate cancer risk in the MCC-Spain case-control study.

Night shift work has been classified as a probable human carcinogen based on experimental studies and limited human evidence on breast cancer. Evidence on other common cancers, such as prostate cancer, is scarce. Chronotype is an individual characteristic that may relate to night work adaptation. We evaluated night shift work with relation to prostate cancer, taking into account chronotype and disease severity in a population based case-control study in Spain. We included 1,095 prostate cancer cases and 1,388 randomly selected population controls. We collected detailed information on shift schedules (permanent vs. rotating, time schedules, duration, frequency), using lifetime occupational history. Sociodemographic and lifestyle factors were assessed by face-to-face interviews and chronotype through a validated questionnaire. We used unconditional logistic regression analysis adjusting for potential confounders. Subjects who had worked at least for one year in night shift work had a slightly higher prostate cancer risk [Odds Ratio (OR) 1.14; 95%CI 0.94, 1.37] compared with never night workers; this risk increased with longer duration of exposure (≥ 28 years: OR 1.37; 95%CI 1.05, 1.81; p-trend = 0.047). Risks were more pronounced for high risk tumors [D'Amico classification, Relative Risk Ratio (RRR) 1.40; 95%CI 1.05, 1.86], particularly among subjects with longer duration of exposure (≥28 years: RRR 1.63; 95%CI 1.08, 2.45; p-trend = 0.027). Overall risk was higher among subjects with an evening chronotype, but also increased in morning chronotypes after long-term night work. In this large population based study, we found an association between night shift work and prostate cancer particularly for tumors with worse prognosis.

Clinically recognizable error rate after the transfer of comprehensive chromosomal screened euploid embryos is low.

OBJECTIVE: To determine the clinically recognizable error rate with the use of quantitative polymerase chain reaction (qPCR)-based comprehensive chromosomal screening (CCS). DESIGN: Retrospective study. SETTING: Multiple fertility centers. PATIENT(S): All patients receiving euploid designated embryos. INTERVENTION(S): Trophectoderm biopsy for CCS. MAIN OUTCOME MEASURE(S): Evaluation of the pregnancy outcomes following the transfer of qPCR-designated euploid embryos. Calculation of the clinically recognizable error rate. RESULT(S): A total of 3,168 transfers led to 2,354 pregnancies (74.3%). Of 4,794 CCS euploid embryos transferred, 2,976 gestational sacs developed, reflecting a clinical implantation rate of 62.1%. In the cases where a miscarriage occurred and products of conception were available for analysis, ten were ultimately found to be aneuploid. Seven were identified in the products of conception following clinical losses and three in ongoing pregnancies. The clinically recognizable error rate per embryo designated as euploid was 0.21% (95% confidence interval [CI] 0.10-0.37). The clinically recognizable error rate per transfer was 0.32% (95% CI 0.16-0.56). The clinically recognizable error rate per ongoing pregnancy was 0.13% (95% CI 0.03-0.37). Three products of conception from aneuploid losses were available to the molecular laboratory for detailed examination, and all of them demonstrated fetal mosaicism. CONCLUSION(S): The clinically recognizable error rate with qPCR-based CCS is real but quite low. Although evaluated in only a limited number of specimens, mosaicism appears to play a prominent role in misdiagnoses. Mosaic errors present a genuine limit to the effectiveness of aneuploidy screening, because they are not attributable to technical issues in the embryology or analytic laboratories.

Markers of oxidative status in a clinical model of oxidative assault: a pilot study in human blood following doxorubicin administration.

We used doxorubicin-based chemotherapy as a clinical model for oxidative assault. Study recruited 23 breast cancer patients and collected blood samples before (T0), at 1 (T1) and 24 hours (T24) after treatment administration. Measurements included protein carbonyl content (PPCC), malondialdehyde (MDA), and alpha- and gamma-tocopherols in plasma and total glutathione content in erythrocytes (erGSHt). In all subjects, PPCC and MDA levels did not change. erGSHt levels increased at T24 by 8% (p=0.03). Levels of alpha, gamma, and total tocopherols progressively decreased by 7%-15% (p <0.05). In subjects with low erGSHt levels (below median), PPCC mean levels progressively increased from 0.35 (T0) to 0.56 (T1) and 0.72 nmol carbonyl/mg protein (T24) (p =0.2). These results indicate that (1) plasma MDA is not a sensitive biomarker in humans; (2) PPCC potentially may be used, if antioxidant reserves are taken into account; (3) antioxidant reserves play an important role in the reaction to oxidative stress.

[Metanephric adenoma. Case report]. / Adenoma metanéfrico. Presentación de un caso.

OBJECTIVE: Metanephric adenoma is an unfrequent renal neoplasm, usually asymptomatic. Differential diagnosis with other renal tumors has to be performed. METHODS/RESULTS: We report the case of a 67 years old woman with a metanephric adenoma. CONCLUSIONS: Metanephric tumors comprise a pathologic spectrum of lesions. At one end of the pathologic spectrum are the metanephric adenomas. These tumors usually behave in a benign fashion. It is important to differentiate metanephric adenoma from clinically aggressive renal tumors.

[Papillary urothelial carcinoma of the urachus]. / Carcinoma papilar urotelial de uraco.

OBJECTIVE: Persistence of a patent urachus is a relatively rare occurrence; however; the persistence of urachal remnants is common. The pathologic lesions of the urachus include benign and malignant neoplasms. The majority of malignant urachal neoplasms are adenocarcinomas. METHODS AND RESULTS: We report a case of papillary urothelial carcinoma of urachus in a 73-year-old woman. The patient was treated with complete surgical extirpation and partial cystectomy. CONCLUSION: The recognition of urachal remnants as vestigial structures and pathologic lesions of the urachus is important.

Endocrinology of recurrent pregnancy loss.

Following implantation, the maintenance of the pregnancy is dependent on a multitude of endocrinological events that will eventually aid in the successful growth and development of the fetus. Although the great majority of pregnant women have no pre-existing endocrine abnormalities, a small number of women can have certain endocrine alterations that could potentially lead to recurrent pregnancy losses. It is estimated that approximately 8 to 12% of all pregnancy losses are the result of endocrine factors. During the preimplantation period, the uterus undergoes important developmental changes stimulated by estrogen, and more importantly, progesterone. Progesterone is essential for the successful implantation and maintenance of pregnancy. Therefore, disorders related to inadequate progesterone secretion by the corpus luteum are likely to affect the outcome of the pregnancy. Luteal phase deficiency, hyperprolactinemia, and polycystic ovarian syndrome are some examples. Several other endocrinological abnormalities such as thyroid disease, hypoparathyroidism, uncontrolled diabetes, and decreased ovarian reserve have been implicated as etiologic factors for recurrent pregnancy loss.

Validity of cuff-uroflow as a diagnostic technique for bladder outlet obstruction in males.

OBJECTIVE: To investigate the validity of the cuff-uroflow method as a diagnostic technique for bladder outlet obstruction (BOO) in males. MATERIAL AND METHODS: A cross-sectional study was carried out on a consecutive series of 93 males (mean age 54.1 years) referred on presenting with lower urinary tract symptoms. The patients were subjected to measurement of the isovolumetric pressure (Pcuff.op) and its corresponding flow (Qcuff.op) by means of the cuff-uroflow method. Subsequently, a study was carried out of the conventional pressure-flow, calculating the degree of BOO according to the provisional International Continence Society (ICS) nomogram. RESULTS: There was a statistical relationship between the Pcuff.op and Qcuff.op values and the degree of BOO according to the provisional ICS nomogram: patients with BOO had a lower value of Qcuff.op and a higher value of Pcuff.op than those without BOO. Using a logistic regression model a nomogram was constructed to link Pcuff.op with Qcuff.op. This nomogram was divided into three areas (obstructed, indeterminate and not obstructed). Most patients (44.1% of the sample) were situated in the indeterminate area. If those patients were excluded, the nomogram gave a sensitivity for the diagnosis of obstruction of 100% and a specificity of 55.6%. The overall rate of correct diagnosis was 84.6%. CONCLUSIONS: The parameters obtained using the cuff-uroflow method were related to the presence of BOO. According to the provisional ICS nomogram, this method showed a better sensitivity than specificity for the diagnosis of BOO.

Reproducibility of non-invasive urodynamics, using the cuff-uroflow, for the diagnosis of bladder outlet obstruction in males.

OBJECTIVE: To determine the repeatability of the parameters obtained from non-invasive urodynamics, using the cuff-uroflow, for the diagnosis of bladder outlet obstruction. MATERIAL AND METHODS: The study was carried out in a consecutive series of 34 males with functional urinary tract symptoms. The test-retest reproducibility of isometric pressure, flow in response to isovolumetric pressure and the energy transfer ratio obtained at two different times using the cuff-uroflow method was determined. RESULTS: Very good agreement for the flow in response to isovolumetric pressure measurement (intraclass correlation coefficient 0.96) and good agreement for the isovolumetric pressure measurement (intraclass correlation coefficient 0.87) and the energy transfer ratio (intraclass correlation coefficient 0.84) were demonstrated. The patients were classified into three groups according to the value of the energy transfer ratio, and it was found that there was very good agreement between the groups into which patients were classified as a result of the first and second measurements (kappa index 0.81). CONCLUSIONS: The parameters obtained with the cuff-uroflow are reliable and the energy transfer ratio allows one to classify patients into reproducible groups.