Multiple neonicotinoids in children’s cerebro-spinal fluid, plasma, and urine

Background Neonicotinoids (NN) are selective neurotoxic pesticides that bind to insect but also mammal nicotinic acetycholine receptors (nAChRs). As the most widely used class of insecticides worldwide, they are ubiquitously found in the environment, wildlife, and foods, and thus of special concern for their impacts on the environment and human health. nAChRs are vital to proper brain organization during the prenatal period and play important roles in various motor, emotional, and cognitive functions. Little is known on children’s contamination by NN. In a pilot study we tested the hypothesis that children’s cerebro-spinal fluid (CSF) can be contaminated by NN. Methods NN were analysed in leftover CSF, blood, and urine samples from children treated for leukaemias and lymphomas and undergoing therapeutic lumbar punctions. We monitored all neonicotinoids approved on the global market and some of their most common metabolites by ultra-high performance liquid chromatography-tandem mass spectrometry. Results From August to December 2020, 14 children were consecutively included in the study. Median age was 8 years (range 3–18). All CSF and plasma samples were positive for at least one NN. Nine (64%) CSF samples and 13 (93%) plasma samples contained more than one NN. Thirteen (93%) CSF samples had N-desmethyl-acetamiprid (median concentration 0.0123, range 0.0024–0.1068 ng/mL), the major metabolite of acetamiprid. All but one urine samples were positive for ≥ one NN. A statistically significant linear relationship was found between plasma/urine and CSF N-desmethyl-acetamiprid concentrations. Conclusions We have developed a reliable analytical method that revealed multiple NN and/or their metabolites in children’s CSF, plasma, and urine. Our data suggest that contamination by multiple NN is not only an environmental hazard for non-target insects such as bees but also potentially for children.

Candidate microRNA Biomarkers in Lupus Nephritis: A Meta-Analysis of Profiling Studies in Kidney, Blood and Urine Samples

Lupus nephritis (LN) is a kidney disease caused by systemic lupus erythematosus in which kidneys are attacked by the immune system. MiRNAs participate in the pathogenesis of LN as modulatory effectors in immune responses and nephrogenesis pathways. Despite the large number of miRNAs that are expressed deferentially, identifying the most suitable biomarker candidate for LN is challenging. The aim of this study was to introduce a consensus panel of potential miRNA biomarkers by performing a meta-analysis of miRNA profiles in the kidney, blood, and urine samples of LN patients. A comprehensive literature review approach was considered to find LN-related miRNA expression profiles. After including the 41 eligible studies and performing the data extraction, meta-analysis was done based on the vote-counting rank strategy and as well as meta-analysis of p-value. The result of the meta-analysis was three lists of consensus miRNAs with altered expression profiles in the various tissue samples of LN patients. Of the 13 studies on kidney tissue, the meta-miRNAs were let-7a, miR-198, let-7e, miR-145, and miR-26a. In addition, meta-miRNAs of miR-199a, miR-21, miR-423, miR-1260b, miR-589, miR-150, miR-155, miR-146a, and miR-183 from 21 studies on blood samples, and miR-146a, miR-204, miR-30c, miR-3201, and miR-1273e from 11 studies on urine samples can be considered as non-invasive biomarker panels for LN. The meta-miRNAs and their related genes were subjected to functional enrichment analyses and network construction. Functional enrichment analysis confirmed the involvement of their target genes in nephropathy-related signaling pathways. Network construction showed miR-145-5p in blood and miR-155-5p in kidney, the best hob miRNAs as candidate drug targets. In conclusion using a meta-analysis approach, our study proposes three meta-miRNA panels that could be the target of further research to assess their potential as biomarkers / therapeutic targets in LN disease.

Liquid biopsy of extracellular vesicle biomarkers for prostate cancer personalized treatment decision

Liquid biopsy of tumor-derived extracellular vesicles (EVs) has great potential as a biomarker source for prostate cancer (CaP) early diagnosis and predicting the stages of cancer. The contents of EVs play an important role in intercellular communication and have specific expression in blood and urine samples from CaP patients. Powered by high-throughput, next-generation sequencing and proteomic technologies, novel EV biomarkers are easily detected in a non-invasive manner in different stages of CaP patients. These identified potential biomarkers can be further validated with a large sample size, machine learning model, and other different methods to improve the sensitivity and specificity of CaP diagnosis. The EV-based liquid biopsy is a novel and less-invasive alternative to surgical biopsies which would enable clinicians to potentially discover a whole picture of tumor through a simple blood or urine sample. In summary, this approach holds promise for developing personalized medicine to guide treatment decisions precisely for CaP patients.

COMPARISON OF A RAPID TEST WITH bPAG ELISA IN PREGNANCY DIAGNOSIS IN COWS

Early pregnancy detection is vital for properly managing livestock farms by re-inseminating nonpregnant females and minimizing the calving intervals. The present investigation was executed to compare a rapid test (Dairy Cow Pregnancy Test from Span Biotech Ltd. Shenzhen, China) with a commercial bPAG ELISA test (IDEXX®) for pregnancy diagnosis in non-descriptive cows. The study also aimed to elucidate if the rapid test could be an alternative method to ELISA in the field. Blood and urine samples were collected from 43 cows to measure the concentrations of bovine pregnancy-associated glycoproteins (bPAGs) in the blood samples and detect progesterone via the rapid test in the urine samples. Examining the genital tracts was achieved after slaughtering the cows to determine the uterus’ state, used as the reference standard for both tests. The results showed that the bPAG ELISA test was more accurate in pregnancy detection than the rapid test. However, there were no statistically significant differences (P > 0.05) between both methods’ results. We conclude that the rapid test can be a suitable alternative method to the bPAG ELISA test for diagnosing pregnancy in cows in field conditions.

Taiwan Biobank: a rich biomedical research database of the Taiwanese population

The Taiwan Biobank (TWB) is an ongoing prospective study of over 150,000 individuals aged 30-70 recruited from across Taiwan beginning in 2012. A comprehensive list of phenotypes was collected for each consented participant at recruitment and follow-up visits through structured interviews and physical measurements. Biomarkers and genetic data were also generated for all participants from blood and urine samples. We present here an overview of the genetic data quality, population structure, and familial relationship within TWB, which consists of predominantly Han Chinese-ancestry individuals, and highlight important attributes and genetic findings thus far from the biobank. A linkage to Taiwan's National Health Insurance database of >25 years and other health-related registries is underway that will enrich the phenotypic spectrum of TWB and enable deep and longitudinal genetic investigations. TWB provides one of the largest biobank resources for biomedical and public health research in East Asia that will contribute to our understanding of the genetic basis of human health and disease in global populations through collaborative studies with other biobanks.

Magyarországra behurcolt trópusi arbovírusfertőzések 2016 és 2020 között

Összefoglaló. Bevezetés: A Dengue-, Zika- és Chikungunya-vírus-fertőzések a trópusokról importált leggyakoribb arbovírusfertőzések. Földrajzi elterjedésük átfedő, közös vektoraik és hasonló tüneteik miatt szerológiai és molekuláris módszerek együttes alkalmazásán alapuló mikrobiológiai vizsgálatokkal különíthetők el megbízhatóan. Célkitűzés: Munkánk célja a 2016 és 2020 között endémiás területen járt, tünetes és tünetmentes utazók vizsgálata volt, minden esetben mindhárom vírusfertőzés irányában. A diagnosztikus tesztek során az alvadásgátolt teljes vér és vizelet bevonásával vizsgáltuk a vírus-RNS kimutathatóságának esélyét a különböző mintatípusokból. Módszer: Savópárminták szerológiai analízise során a Dengue-, Zika- és Chikungunya-vírus-specifikus ellenanyagválasz alakulását vizsgáltuk ELISA-módszerrel. Reaktív eredmények esetében a szerológiai keresztreakciók kizárására immunfluoreszcens és ELISA-technikán alapuló további vizsgálatokat végeztünk a hazai és az utazás során érintett területeken előforduló flavi- és alphavirusok irányában. Vérsavó-, alvadásgátolt teljes vér és vizeletmintákból reverztranszkripciót követő valós idejű polimeráz-láncreakcióval vírus-RNS-kimutatást végeztünk. Eredmények: Az 1037 vizsgált utazó közül 133 esetben kaptunk reaktív szerológiai és/vagy molekuláris eredményt. Az alvadásgátolt teljes vér mintából sikerült a legnagyobb arányban vírusnukleinsavat kimutatni mind a Dengue- és Zika-, mind a Chikungunya-vírus esetében. Megbeszélés: Endémiás területről hazatért utazók vizsgálatát a tünetek hasonlósága miatt mindhárom vírusfertőzés irányában együttesen indokolt elvégezni. A flavi- és alphavirusokra jellemző nagyfokú szerológiai keresztreaktivitás miatt a nukleinsav-kimutatás javíthatja a mikrobiológiai diagnosztika pontosságát. Következtetés: A három vírus mikrobiológiai diagnosztikáját segíti a korai mintavétel és a molekuláris vizsgálatok kiterjesztése további mintatípusokra: alvadásgátolt teljes vér és vizelet. A behurcolt vírusfertőzések azonosítása fokozott jelentőségű, mert az Európában is jelen lévő vektorszúnyogfajok felvetik az autochton átvitel lehetőségét. Orv Hetil. 2021; 162(50): 2000–2009. Summary. Introduction: Dengue-, Zika- and Chikungunya infections are among the most frequently imported tropical arbovirus infections. Due to their shared endemic regions, vectors and similar clinical symptoms, differential diagnosis is based on serological and molecular analysis. Objective: The aim of our study was to identify the imported arbovirus infections of travellers between 2016 and 2020. Furthermore, to improve the diagnostic sensitivity, anticoagulated whole blood and urine samples were involved in molecular diagnosis. Method: Virus-specific antibody kinetics was tested in paired sera of patients by ELISA method. In case of reactive results, further serological analysis was performed using immunofluorescence assays and/or ELISA tests to exclude serological cross-reactions caused by other members of the flavi- and alphaviruses. Detection of viral RNA was attempted from serum, anticoagulated whole blood and urine specimens using reverse transcription and real-time polymerase chain reaction. Results: Out of the tested 1037 travellers, reactive serological and/or molecular results were obtained in 133 cases. Anticoagulated whole blood proved to be the most suitable specimen for viral RNA detection of the three viruses. Discussion: Parallel testing of Dengue-, Zika- and Chikungunya infections is recommended, as symptom-based differential diagnosis is challenging. Due to the characteristic serological cross-reactivity of flavi- and alphaviruses, microbiological diagnosis relies on both serological and molecular tests. Conclusion: Involving anticoagulated whole blood and urine samples into molecular analysis and early sample collection improve the sensitivity of microbiological diagnostics. Identification of imported tropical arbovirus infections is of high importance as the presence of vector mosquitos in Europe raises the possibility of autochthon transmission. Orv Hetil. 2021; 162(50): 2000–2009.

First Isolation and Molecular Typing of Pathogenic and Intermediate Leptospira Species from Urine of Symptomatic Dogs

Aim of this study was to evaluate, the presence and diversity of Leptospira spp. in blood and urine samples collected from 175 owned-dogs from Sardinia, Italy. After determination of leptospiral infection by microscopic agglutination test (MAT), urine from MAT-positive dogs were examined by real-time polymerase chain reaction (lipL32 rt-PCR) and then isolated by culture. In order to characterize obtained serovars, positive cultures were then subjected to 16S rRNA and secY sequencing, phylogenetic analysis and Multilocus Sequence Typing (MLST). Results showed that seven dogs (4%; 95% CI: 0–55) had Leptospira DNAs in their urine and five strains were isolated from urine cultures. The three different sequence types (ST17, ST198 and ST24) belonging to Leptospira interrogans genomospecies identified by MLST analyses in this study, confirmed that the leptospiral infection was widespread in Sardinian dogs. We also reported the first characterization of a new Leptospira spp. isolated from urine of one dog living in the study area. Whole genome sequencing and phylogenetic analysis, confirmed that this genospecies was closely related to Leptospira hovindhougenii, an intermediate Leptospira spp. with unknown pathogenicity previously isolated from a rat in Denmark. Further studies are required to clarify whether healthy dogs that shed leptospires in their urine could represent a zoonotic risk for humans in this region.

Silicon Resorption from Equisetum arvense Tea – A Randomized, Three-Armed Pilot Study

Equisetum arvense tea (TEA) contains high concentrations of silicon and has been used in folk medicine for the treatment of inflammatory ailments. We examined the resorption of silicon after TEA consumption. Safety and immunological effects were secondary outcomes. A monocentric, randomized, three-armed pilot study was conducted with 12 voluntary, healthy, male subjects. The study is registered in the German register for clinical trials (DRKS-ID: DRKS00016628). After a low silicon diet for 36 hours, 1000 mL TEA1 with approximately 200 000 µg silicon/L, TEA2 with approximately 750 000 µg silicon/L, or Si-low-Water (approximately 10 – 10 000 µg silicon/L as a control) were ingested on three consecutive days. Blood and urine samples were collected at baseline, day 1 examining silicon kinetics, day 3 examining silicon accumulation, and day 8 (safety, immunological parameters). Si-low-Water intake did not change silicon serum (Cmax 294 µg/L) or urine (19 000 µg/24 h) concentrations compared to baseline. Cmax was 2855 µg/L for TEA1 and 2498 µg/L for TEA2; tmax was 60 and 120 min, respectively. Silicon accumulation did not occur. Urine silica within 24 h (E24 h) was higher after TEA2 compared to TEA1 ingestion (142 000 vs. 109 000 µg/24 h). Serum silicon levels at t = 120 min differed significantly after intake of TEA2 or intake of Si-low-Water (p = 0.029). The immunological parameters did not show any significant changes indicating immunosuppressive effects in volunteers. TEA1 was well tolerated, while TEA2 caused diarrhoea in 4 subjects. Our investigations show that intake of TEA1 leads to significant rise in serum silicon concentration.

Fractional electrolyte excretion and osmolality in the early diagnosis of acute kidney injury in cats with urethral obstruction

Acute Kidney Injury (AKI) can be defined as a spectrum of diseases associated with a sudden onset of a renal failure status, the feline patient has azotemia, the disorder in fractional electrolyte excretion (FE), and shedding of epithelial cells from renal tubular segments observed in the urinary sediment. Thus, the objective of the present study was to evaluate the FE, plasma and urinary osmolality, and urinary specific gravity (USG) in cats that spontaneously developed AKI due to urethral obstruction (UO) and healthy cats. Blood and urine samples were collected from a group of 20 cats diagnosed with AKI secondary to urethral obstruction (GAKI; n=20) and clinically healthy cats (GC; n=15). The serum creatinine (sCre) and urinary creatinine (uCre), were measured by spectrophotometry, serum and urinary analyzes of sodium, potassium and chloride by ion selective electrode device and serum and urinary osmolarity by osmometer. The GAKI results were statistically compared with those of the CG using Student's t tests to assess normal data, while the Maan-Whitney test was used for non-normal data. A significant increase in the sCr, sK, FENa, FECl and RFI parameters of the GAKI cats when compared to the GC (p < 0.05). The sCl, USG, uCr, uK and uOSM parameters decreased significantly when compared between the two groups. Thus, given the established methodology and the results found, it is possible to infer that an increase in EFNa, EFCl in addition to the RFI and a decrease in USG and uOSM were associated with cats with AKI and can serve as markers of kidney damage, as well as monitoring the prognosis.