Danielle de Jong

Introduction: Schistosomiasis remains a major public health challenge, particularly in sub-Saharan Africa. Detecting low-intensity infections is difficult because conventional parasitological methods lack sufficient sensitivity. Circulating anodic antigen (CAA), a genus-specific biomarker released by Schistosoma worms, enables detection of active Schistosoma infections. However, existing CAA assays require laboratory-based sample processing. We evaluated Flow-S, a field-deployable device for high-sensitivity quantitative CAA detection directly from large-volume, untreated urine samples. Flow-S integrates CAA concentration and quantitative upconverting reporter particle lateral flow (UCP-LF) detection with a portable standalone battery-powered reader for field-use. Methods: A prospective study was conducted by local healthcare staff in a remote Schistosoma haematobium -endemic low-to-moderate infection setting. Fresh urine samples (n = 158) from women of reproductive age were analyzed using Flow-S. Paired urine, serum, and stool samples were examined by urine filtration microscopy, Kato-Katz (to exclude S. mansoni ), and laboratory-based UCP-LF CAA reference assays (serum SCAA20 and urine UCAA hT 417). A composite reference standard (CRS) classified individuals as infected when eggs or either reference CAA assay was positive. Flow-S performance was evaluated against the CRS and further assessed using latent class analysis (LCA). Results: Complete datasets were available for 115 samples. Flow-S achieved an area under the receiver operating characteristic (ROC) curve of 0.92 and detected all egg-positive infections. Using a positivity threshold of 1.10 pg/mL, Flow-S achieved 80.8% sensitivity and 96.6% specificity, meeting the World Health Organization ideal target product profile requirements for monitoring and evaluation of schistosomiasis control programs. Latent class analysis provided complementary model-based evidence consistent with the high discriminatory performance of Flow-S observed using the composite reference standard. Conclusions: Flow-S enables quantitative, field-deployable detection of S. haematobium infections directly from untreated large-volume urine without laboratory pre-treatment. Its high analytical sensitivity and diagnostic performance support its potential for detecting low-intensity infections. At an operational threshold of 1.10 pg/mL, Flow-S met the WHO TPP ideal targets for monitoring and evaluation and showed potential for cost-effective surveillance strategies, including pooled-sample screening for schistosomiasis control and elimination programs.

We quantified incident pregnancies among reproductive-aged women with and without Schistosoma haematobium infection. Among 245 women, 59 (24.1%) became pregnant during a median follow-up of 370 days. Women with S. haematobium infection had a 52% lower rate of pregnancy within the study period compared with uninfected women (hazard ratio = 0.48 [0.29–0.80], P = .005).

: Latent tuberculosis infection (LTBI) and Schistosoma mansoni are common in Africa, and helminth-induced immunomodulation may affect LTBI detection. This study aimed to assess whether S. mansoni infection affects LTBI detection by the QuantiFERON-TB Gold Plus (QFT-Plus) assay and alters serum interferon-γ (IFN-γ) concentrations in response to Mycobacterium tuberculosis ( Mtb ) antigens at baseline and after 1 year, during which participants with S. mansoni infection received praziquantel treatment. At baseline, 65 individuals with schistosome infection had lower average IFN-γ concentrations in TB1-stimulated QFT-Plus supernatants compared with 83 uninfected individuals (10.4 versus 51.9 pg/mL, P = 0.038). Although not statistically significant, QFT-Plus test positivity rate was unexpectedly slightly higher among adults with schistosome infection at baseline (26.2% versus 18.1%, P = 0.24). The incidence over 12 months was higher posttreatment in participants initially infected with S. mansoni compared with those uninfected (13.9% [ n = 5/36] versus 4.2% [ n = 2/48], P = 0.13). By 12 months, IFN-γ concentrations were comparable between the two groups (53.8 versus 33.5 pg/mL, respectively, P = 0.56). Individuals who cleared S. mansoni infection experienced a nearly 12-fold increase in IFN-γ levels relative to those who remained uninfected, although this difference did not reach statistical significance ( P = 0.17). In conclusion, baseline S. mansoni infection was associated with suppressed IFN-γ responses to Mtb antigens, suggesting helminth-induced immune dampening. Praziquantel treatment may partially restore TB-specific immune responses and facilitate LTBI detection. These findings highlight the potential role of S. mansoni as a critical cofactor affecting LTBI diagnosis in schistosomiasis-endemic regions.

A laboratory-based lateral flow (LF) test that utilizes up-converting reporter particles (UCP) for ultrasensitive quantification of Schistosoma circulating anodic antigen (CAA) in urine is a well-accepted test to identify active infection. However, this UCP-LF CAA test requires sample pre-treatment steps not compatible with field applications. Flow, a new low-cost disposable, allows integration of large-volume pre-concentration of urine analytes and LF detection into a single field-deployable device. We assessed a prototype Flow-Schistosoma (Flow-S) device with an integrated UCP-LF CAA test strip, omitting all laboratory-based steps, to enable diagnosis of active Schistosoma infection in the field using urine. Flow-S is designed for large-volume (5–20 mL) urine, applying passive paper-based filtration and antibody-based CAA concentration. Samples tested for schistosome infection were collected from women of reproductive age living in a Tanzania region where S. haematobium infection is endemic. Fifteen negative and fifteen positive urine samples, selected based on CAA levels quantified in paired serum, were analyzed with the prototype Flow-S. The current Flow-S prototype, with an analytical lower detection limit of 1 pg CAA/mL, produced results correlated with the laboratory-based UCP-LF CAA test. Urine precipitates occurred in frozen banked samples and affected accurate quantification; however, this should not occur in fresh urine. Based on the findings of this study, Flow-S appears suitable to replace the urine pre-treatment required for the laboratory-based UCP-LF CAA test, thus allowing true field-based applications with fresh urine samples. The urine precipitates observed with frozen samples, though less important given the goal of testing fresh urines, warrant additional investigation to evaluate methods for mitigation. Flow-S devices permit testing of pooled urine samples with applications for population stratified testing. A field test with fresh urine samples, a further optimized Flow-S device, and larger statistical power has been scheduled.

The tumor suppressor protein p53 has an important role in cell-fate determination. In cancer cells, the activity of p53 is frequently repressed by high levels of MDMX and/or MDM2. MDM2 is a ubiquitin ligase whose activity results in ubiquitin- and proteasome-dependent p53 degradation, while MDMX inhibits p53-activated transcription by shielding the p53 transactivation domain. Interestingly, the oncogenic functions of MDMX appear to be more wide-spread than inhibition of p53. The present study aimed to elucidate the MDMX-controlled transcriptome. Therefore, we depleted MDMX with four distinct shRNAs from a high MDMX expressing uveal melanoma cell line and determined the effect on the transcriptome by RNAseq. Biological function analyses indicate the inhibition of the cell cycle regulatory genes and stimulation of cell death activating genes upon MDMX depletion. Although the inhibition of p53 activity clearly contributes to the transcription regulation controlled by MDMX, it appeared that the transcriptional regulation of multiple genes did not only rely on p53 expression. Analysis of gene regulatory networks indicated a role for Forkhead box (FOX) transcription factors. Depletion of FOXO proteins partly prevented the transcriptional changes upon MDMX depletion. Furthermore, depletion of FOXO proteins relatively diminished the growth inhibition upon MDMX knockdown, although the knockdown of the FOXO transcription factors also reduces cell growth. In conclusion, the p53-independent oncogenic functions of MDMX could be partially explained by its regulation of FOXO activity.

Melanoma is a leading cause of high mortality that frequently spreads to the brain and is associated with deterioration in quality and quantity of life. Treatment opportunities have been restricted until now and new therapy options are urgently required. Our focus was to reveal the potential heterogeneity of melanoma brain metastasis. We succeeded to establish a brain melanoma metastasis cell line, namely MUG-Mel1 and two resulting clones D5 and C8 by morphological variety, differences in lipidome, growth behavior, surface, and stem cell markers. Mutation analysis by next-generation sequencing, copy number profiling, and cytogenetics demonstrated the different genetic profile of MUG-Mel1 and clones. Tumorigenicity was unsuccessfully tested in various mouse systems and finally established in a zebra fish model. As innovative treatment option, with high potential to pass the blood-brain barrier a peptide isolated from lactoferricin was studied in potential toxicity. Brain metastases are a major clinical challenge, therefore the development of relevant in vitro and in vivo models derived from brain melanoma metastases provides valuable information about tumor biology and offers great potential to screen for new innovative therapies.

Nature Communications 7: Article number: 13874 (2016); Published 22 December 2016; Updated 6 September 2017 The financial support for this Article was not fully acknowledged. The Acknowledgements should have included the National Institutes of Health grants R01GM098749 and National Institutes of Health Transformative Research Award R01NS096786.

In humans, the defective invasion of the maternal endometrium by fetal extravillous trophoblasts (EVTs) can lead to insufficient perfusion of the placenta, resulting in pregnancy complications that can put both mother and baby at risk. To study the invasion of maternal endometrium between (W)5.5–12 weeks of gestation by EVTs, we combined fluorescence in situ hybridization, immunofluorescence and immunohistochemistry to determine the presence of (male) EVTs in the vasculature of the maternal decidua. We observed that interstitial mononuclear EVTs directly entered decidual veins and lymphatics from W5.5. This invasion of decidual veins and lymphatics occurred long before endovascular EVTs remodelled decidual spiral arteries. This unexpected early entrance of interstitial mononuclear EVTs in the maternal circulation does not seem to contribute to the materno-placental vascular connection directly, but rather to establish (and expand) the materno-fetal interface through an alternative vascular route.