Assessment of the exposure of breast milk to persistent organic pollutants in Latvia

According to the National Implementation Plan on persistent organic pollutants (POP) for the years 2004-2020 the situation on POP was actualised in Latvia. The aim of the study was to assess the contamination of breast milk with polychlorinated biphenyls (PCB), polychlorinated dibenzodioxins, dibenzofurans (PCDD/PCDF) and some pesticides. Breast feeding mothers (together 30) were selected from probably polluted area, and a control group on the basis of WHO criteria for donor selection. The concentration of POP in milk was detected by gas chromatography with HRGC/HRMS, HRGC/ECD and GC/ECD detection after extraction of milk fat and purification. The results of the survey show comparability of both groups in age, weight, habits, occupation as well as age of the children and the weight at birth and at the time of collecting the samples. PCB were not observed in 15.9%, PCDD/PCDF in 6.6% and pesticides in 14% analyses of the target group and in 26.3%, 7% and 8.6% analyses correspondingly of the control group. The concentration of 18 detected PCB, 17 detected PCDD/PCDF and 8 chlorinated pesticides and its metabolites varied in wide range and the differences of mean values within groups were not statistically significant (p>0.05). The concentration of POPs corresponds to lowest levels detected in European countries. The concentration of dioxine-like PCBs and PCDD/PCDF according to toxicity equivalents does not exceed the WHO accepted level of 15 TEQ pg/g milk fat.     

High-throughput detection of pathogenic yeasts of the genus trichosporon

The need for a rapid and accurate method for the detection of fungal pathogens has become imperative as the incidence of fungal infections has increased dramatically. Herein, we tested the Luminex 100, a novel flow cytometer, for the detection of the medically important genus Trichosporon. This genus was selected as our proof-of-concept model due to the close phylogenetic relationship between the species. The method, which is based on a nucleotide hybridization assay, consists of a combination of different sets of fluorescent beads covalently bound to species-specific capture probes. Upon hybridization, the beads bearing the target amplicons are classified by their spectral addresses with a 635-nm laser. Quantitation of the hybridized biotinylated amplicon is based on fluorescence detection with a 532-nm laser. We tested in various multiplex formats 48 species-specific and group-specific capture probes designed in the D1/D2 region of ribosomal DNA, internal transcribed spacer regions, and intergenic spacer region. Species-specific biotinylated amplicons were generated with three sets of primers to yield fragments from the three regions. The assay was specific and fast, as it discriminated species differing by 1 nucleotide and required less than 50 min following amplification to process a 96-well plate. The sensitivity of the assay allowed the detection of 10(2) genome molecules in PCRs and 10(7) to 10(8) molecules of biotinylated amplification product. This technology provided a rapid means of detection of Trichosporon species with the flexibility to identify species in a multiplex format by combining different sets of beads.     

Near universal detection of alterations in CTNNB1 and Wnt pathway regulators in desmoid‐type fibromatosis by whole‐exome sequencing and genomic analysis

CTNNB1 mutations or APC abnormalities have been observed in ～85% of desmoids examined by Sanger sequencing and are associated with Wnt/β‐catenin activation. We sought to identify molecular aberrations in “wild‐type” tumors (those without CTNNB1 or APC alteration) and to determine their prognostic relevance. CTNNB1 was examined by Sanger sequencing in 117 desmoids; a mutation was observed in 101 (86%) and 16 were wild type. Wild‐type status did not associate with tumor recurrence. Moreover, in unsupervised clustering based on U133A‐derived gene expression profiles, wild‐type and mutated tumors clustered together. Whole‐exome sequencing of eight of the wild‐type desmoids revealed that three had a CTNNB1 mutation that had been undetected by Sanger sequencing. The mutation was found in a mean 16% of reads (vs. 37% for mutations identified by Sanger). Of the other five wild‐type tumors sequenced, two had APC loss, two had chromosome 6 loss, and one had mutation of BMI1. The finding of low‐frequency CTNNB1 mutation or APC loss in wild‐type desmoids was validated in the remaining eight wild‐type desmoids; directed miSeq identified low‐frequency CTNNB1 mutation in four and comparative genomic hybridization identified APC loss in one. These results demonstrate that mutations affecting CTNNB1 or APC occur more frequently in desmoids than previously recognized (111 of 117; 95%), and designation of wild‐type genotype is largely determined by sensitivity of detection methods. Even true CTNNB1 wild‐type tumors (determined by next‐generation sequencing) may have genomic alterations associated with Wnt activation (chromosome 6 loss/BMI1 mutation), supporting Wnt/β‐catenin activation as the common pathway governing desmoid initiation. © 2015 Wiley Periodicals, Inc.     

The Impact of the Loop Electrosurgical Excisional Procedure for Cervical Intraepithelial Lesions on Female Sexual Function

IntroductionVery limited knowledge exists concerning the impact of Loop Electrosurgical Excisional Procedure (LEEP) on female sexual function in women with cervical intraepithelial neoplasiaAimTo investigate sexual function in women who underwent LEEP for the treatment of cervical intraepithelial lesions, using a validated questionnaire (Female Sexual Function Index [FSFI]).Main Outcome MeasuresFSFI questionnaire on six domains of female sexuality (desire, arousal, lubrication, orgasm, satisfaction, and pain).MethodsConsecutive sexually active women, who underwent LEEP for the treatment of cervical intraepithelial neoplasia were enrolled in this study. All women were asked to complete a copy of FSFI questionnaire, at the time of LEEP and after 6 months. We finally compared the results of the pre-LEEP questionnaire and the post-LEEP questionnaire for each patient.ResultsA total of 67 sexually active women undergoing LEEP for the treatment of cervical intraepithelial lesions were enrolled. Nine of these patients (13.4%) completed only the questionnaire regarding their sexual function before LEEP; thus we did not include them for final analysis. In our study population, data showed a sexual function overall unchanged after LEEP; only the variable “desire” (sexual interest) became significantly worse (P = 0.02).ConclusionsLEEP for the treatment of cervical intraepithelial lesions doesn't affect women's sexuality, when compared with sexual function before surgery. In our study, all FSFI sexual function domains but desire, did not show significant change after LEEP. Serati M, Salvatore S, Cattoni E, Zanirato M, Mauri S, Siesto G, Cromi A, Ghezzi F, and Bolis P. The impact of the Loop Electrosurgical Excisional Procedure for cervical intraepithelial lesions on female sexual function.     

Post‐renal transplant erythrocytosis: A case report

PTE is defined as hematocrit >51% or hemoglobin >17 g/dL after renal transplantation. Risk factors include native kidneys with adequate erythropoiesis pretransplant, smoking, renal artery stenosis, and cyclosporine treatment. We report the case of a 14‐yr‐old female kidney transplant patient, with triple therapy immunosuppression and stable graft function who developed PTE at 12 months post‐transplant with hemoglobin 17.3 g/dL, hematocrit 54.2%, stable graft function, and normotensive with normal cardiac echocardiogram and erythropoietin levels. The only risk factor found was tobacco use. As she had no spontaneous improvement, enalapril treatment was started at 19 months post‐transplant with a hemoglobin level of 17.5 g/dL and hematocrit 53%; by 23 months post‐transplant, hemoglobin lowered to 15 g/dL and hematocrit to 44.5% and continued to be in normal range thereafter. PTE is a rare condition in childhood and can be successfully treated with enalapril.     

Italian National Survey of Blood Donors: External Quality Assessment (EQA) of Syphilis Testing

The detection of syphilis among blood donors may reveal high-risk sexual behavior, which can go unreported at the time of donor selection and compromise the safety of the donated blood. In Italy, blood is collected, tested, and distributed by transfusion services (TSs), which also perform outpatient transfusions. Although the TSs must screen for syphilis by law, there are no indications of the specific type of method to be used, generating discrepancies in the results obtained by the different TSs. To determine the proficiency of the TSs in screening for syphilis, we performed an external quality assessment (EQA). The EQA was based on two shipments of serum panels; 133 and 118 of the 326 existing TSs participated in the first and second shipments, respectively. Each panel consisted of both positive and negative serum samples. The results confirmed that the use of a single nontreponemal test (the Venereal Disease Research Laboratory [VDRL] and the rapid plasma reagin [RPR] tests) is the least sensitive means of identifying samples that are positive for syphilis antibodies. We also found that the interpretation of the results of manual techniques, such as the RPR test, the VDRL test, the Treponema pallidum hemagglutination (TPHA) assay, and the T. pallidum particle agglutination (TPPA) assay, can vary greatly among different TSs and operators. Total Ig enzyme immunoassays (EIAs) are the most sensitive. However, the determination of syphilis on the basis of the results of a single test is not sufficient for an accurate screening; and all blood units should thus be assessed by two distinct treponemal tests, that is, a total Ig EIA and the TPHA or the TPPA assay.Syphilis is a reemerging disease and is caused by the spirochete Treponema pallidum. In most cases it is sexually transmitted, although it can also be transmitted from mother to child in utero and, rarely, through blood transfusion, especially through the transfusion of fresh blood components (6, 12). Serological tests for syphilis are considered to be a milestone in syphilis control, and since as early as the 1930s they have greatly contributed to the detection of T. pallidum infection not only in the clinical setting but also in transfusional medicine. The detection of blood donors who are positive for syphilis is an important public health concern, given that testing may reveal high-risk sexual behavior, which can go unreported at the time of donor selection and compromise the safety of blood used for transfusions.In some European countries, the occurrence of T. pallidum infection in the general population has been increasing, and this increase is reflected in its growing occurrence among blood donors. In particular, a survey performed in England has indicated that since 2001 there has been a trend toward a moderate increase in the incidence of T. pallidum infection among blood donors (2). In Germany, although the incidence of infection among donors is very low, increases have been recorded since 1991 (9). The incidence of T. pallidum infection among blood donors is also increasing in Italy. In particular, according to the Transfusion Transmitted Infections Surveillance System (7), it increased from 3.8 per 100,000 donations in 1999 to 7 per 100,000 donations in 2006 (11).The data collected by the Transfusion Transmitted Infections Surveillance System in Italy are provided by the existing 326 transfusion services (TSs). These are hospital-based facilities where blood is collected, tested, and distributed and where, in most cases, outpatient transfusions are performed (10). In 2006, there were 1,539,454 donors, for a total of 2,402,267 donated units. Remarkably, 85% of the donors provided multiple donations (4). Although the TSs are required by law to screen for syphilis, there are no indications of the specific type of method that must be used, nor is there any confirmatory algorithm for testing on the basis of the different assays available. In fact, the laboratory assessment of syphilis is generally based on the detection of antibodies against T. pallidum antigens in blood by the use of either specific or nonspecific reagents. Methods based on the detection of specific Treponema antigens include passive agglutination, such as the T. pallidum hemagglutination (TPHA) assay or the T. pallidum particle agglutination (TPPA) assay, and indirect immunofluorescence, such as the fluorescent treponemal antibody absorbed (FTA-ABS) assay or the most sensitive assay, the enzyme immunoassay (EIA), for the detection of specific IgG and IgM or total Ig. Additional methods are based on nonspecific reagents, including nontreponemal lipid antigens (cardiolipin), and they most commonly rely on the flocculation technique. Of these, the Venereal Disease Research Laboratory (VDRL) and rapid plasma reagin (RPR) tests are the most commonly used.The use of different assays from such a large array could generate discrepancies in the detection of syphilis among TSs, stressing the need for quality assessment. To this end, we performed an external quality assessment (EQA) of the quality and the comparability of the results obtained by the different TSs with the aim of contributing to the development of preventive and corrective measures (1, 3).