Target fishing reveals PfPYK-1 and PfRab6 as potential targets of an antiplasmodial 4-anilino-2-trichloromethylquinazoline hit compound.

We present investigations about the mechanism of action of a previously reported 4-anilino-2-trichloromethylquinazoline antiplasmodial hit-compound (Hit A), which did not share a common mechanism of action with established commercial antimalarials and presented a stage-specific effect on the erythrocytic cycle of P. falciparum at 8 < t < 16 h. The target of Hit A was searched by immobilising the molecule on a solid support via a linker and performing affinity chromatography on a plasmodial lysate. Several anchoring positions of the linker (6,7 and 3') and PEG-type linkers were assessed, to obtain a linked-hit molecule displaying in vitro antiplasmodial activity similar to that of unmodified Hit A. This allowed us to identify the PfPYK-1 kinase and the PfRab6 GTP-ase as potential targets of Hit A.

Epigenomic alterations define lethal CIMP-positive ependymomas of infancy.

Ependymomas are common childhood brain tumours that occur throughout the nervous system, but are most common in the paediatric hindbrain. Current standard therapy comprises surgery and radiation, but not cytotoxic chemotherapy as it does not further increase survival. Whole-genome and whole-exome sequencing of 47 hindbrain ependymomas reveals an extremely low mutation rate, and zero significant recurrent somatic single nucleotide variants. Although devoid of recurrent single nucleotide variants and focal copy number aberrations, poor-prognosis hindbrain ependymomas exhibit a CpG island methylator phenotype. Transcriptional silencing driven by CpG methylation converges exclusively on targets of the Polycomb repressive complex 2 which represses expression of differentiation genes through trimethylation of H3K27. CpG island methylator phenotype-positive hindbrain ependymomas are responsive to clinical drugs that target either DNA or H3K27 methylation both in vitro and in vivo. We conclude that epigenetic modifiers are the first rational therapeutic candidates for this deadly malignancy, which is epigenetically deregulated but genetically bland.

Tail and ear necrosis in piglets of sows with increased weight loss over the suckling period.

INTRODUCTION: A farm belonging to a Swiss sow pool system reported increased cases of necrosis on the base of the tail or ears in their piglets. Therefore, herd examination was performed in February 2021, and it was found that about half of all examined litters included piglets with necrosis of different locations, and that the sows of these piglets were rather thin. Upon instruction, the farmer then documented the body condition score (BCS) and weight before farrowing and after weaning, and the number of liveborn piglets affected by necrosis of the tail or ear of the next four farrowing batches. In total, data of 97 sows with 1214 liveborn piglets were evaluated. Sows were retrospectively allocated into two groups: Those with piglets with ear and/or tail necrosis (NE), and those without (WN). Of the 97 litters, 40 included piglets with necrosis, with 28 of them having piglets only with tail necrosis, 8 only with ear necrosis, and 4 litters included piglets with both types of necrosis. The group NE lost significantly more weight and BCS points over the suckling period than the group WN, with a tendency of having a lower BCS after weaning (2,0 vs. 2,25/5,0). Blood samples of five sows were analyzed and tested positive for the Fusarium mycotoxin deoxynivalenol (DON). It could be possible that the sows previously consumed DON contaminated feed, which was then stored in their fat tissue, and released again into the blood stream during increased weight loss. Since DON can be transferred from the sow to her piglets during gestation or lactation, this release might have affected the piglets, leading to tail or ear necrosis. However, causative studies are needed to confirm this hypothesis.

INTRODUCTION: Une exploitation appartenant à un système suisse de pool de truies a signalé une augmentation des cas de nécrose à la base de la queue ou des oreilles chez ses porcelets. Par conséquent, un examen de l'effectif a été effectué en février 2021 et il a été constaté qu'environ la moitié de toutes les portées examinées comprenaient des porcelets présentant des nécroses à différents endroits et que les mères de ces porcelets étaient plutôt maigres. Sur instruction, l'éleveur a ensuite documenté la note d'état corporel (BCS) et le poids avant la mise bas et après le sevrage, ainsi que le nombre de porcelets nés vivants affectés par une nécrose de la queue ou de l'oreille des quatre lots de mise bas suivants. Au total, les données de 97 truies avec 1214 porcelets nés vivants ont été évaluées. Les truies ont été réparties rétrospectivement en deux groupes : Celles dont les porcelets présentaient une nécrose de l'oreille et/ou de la queue (NE), et celles qui n'en présentaient pas (WN). Sur les 97 portées, 40 comprenaient des porcelets atteints de nécrose, dont 28 uniquement avec une nécrose de la queue, 8 uniquement avec une nécrose de l'oreille et 4 avec les deux types de nécrose. Le groupe NE a perdu beaucoup plus de poids et de points BCS pendant la période d'allaitement que le groupe WN, avec une tendance à avoir un BCS plus faible après le sevrage (2,0 vs. 2,25/5,0). Les échantillons de sang de cinq truies ont été analysés et se sont révélés positifs pour la mycotoxine de Fusarium, le déoxynivalénol (DON). Il est possible que les truies aient consommé des aliments contaminés par le DON qui a ensuite été stocké dans leur tissu adipeux puis libéré dans le sang lors de la perte de poids. Comme le DON peut être transféré de la truie à ses porcelets pendant la gestation ou la lactation, cette libération pourrait avoir affecté les porcelets, entraînant une nécrose de la queue ou des oreilles. Cependant, des études causales sont nécessaires pour confirmer cette hypothèse.

Assessment of exposure to bovine spongiform encephalopathy in a hypothetical country.

The authors present a basic quantitative spreadsheet model to evaluate the risk of bovine spongiform encephalopathy (BSE) within a national setting. The model is based on information from BSE risk assessments undertaken in Latin American countries. The analysis focuses on the level of regulatory implementation and its impact over different time periods and estimates the potential impact if one BSE-infected animal is introduced into the production cycle. The information is consolidated so that the results of the evaluation can be presented for a hypothetical country, 'Country X'. Evaluating the BSE cycle within a country may help in making decisions on where preventive or control measures should be placed and/or enforced. Such an evaluation may also be used as the exposure assessment of a risk assessment, recommended by the World Organisation for Animal Health to determine the BSE risk status of a given country.

A population genomic approach to map recent positive selection in model species.

Based on nearly complete genome sequences from a variety of organisms data on naturally occurring genetic variation on the scale of hundreds of loci to entire genomes have been collected in recent years. In parallel, new statistical tests have been developed to infer evidence of recent positive selection from these data and to localize the target regions of selection in the genome. These methods have now been successfully applied to Drosophila melanogaster, humans, mice and a few plant species. In genomic regions of normal recombination rates, the targets of positive selection have been mapped down to the level of individual genes.

Combining Extracellular miRNA Determination with Microfluidic 3D Cell Cultures for the Assessment of Nephrotoxicity: a Proof of Concept Study.

Drug-induced kidney injury is often observed in the clinics and can lead to long-term organ failure. In this work, we evaluated a novel in vitro system that aims at detecting whether compounds can cause renal proximal tubule damage in man. For this, we implemented organotypic cultures of human conditionally immortalized proximal tubule epithelial cells overexpressing the organic anion transporter 1 (ciPTEC-OAT1) in a three-channel OrganoPlate under microfluidic conditions. Cells were exposed to four known nephrotoxicants (cisplatin, tenofovir, cyclosporine A, and tobramycin). The effect on cell viability and NAG release into the medium was determined. A novel panel of four miRNAs (mir-21, mir-29a, mir-34a, and mir-192) was selected as potential biomarkers of proximal tubule damage. After nephrotoxicant treatment, miRNA levels in culture medium were earlier indicators than cell viability (WST-8 assay) and outperformed NAG for proximal tubule damage. In particular, mir-29a, mir-34a, and mir-192 were highly reproducible between experiments and across compounds, whereas mir-21 showed more variability. Moreover, similar data were obtained in two different laboratories, underlining the reproducibility and technical transferability of the results, a key requirement for the implementation of novel biomarkers. In conclusion, the selected miRNAs behaved like sensitive biomarkers of damage to tubular epithelial cells caused by several nephrotoxicity mechanisms. This biomarker panel, in combination with the 3D cultures of ciPTEC-OAT1 in the OrganoPlate, represents a novel tool for in vitro nephrotoxicity detection. These results pave the way for the application of miRNAs in longitudinal, time-course in vitro toxicity studies.

Author Correction: Yolk sac macrophage progenitors traffic to the embryo during defined stages of development.

This article contains errors in Figs. 5 and 6, for which we apologize. In Fig. 5f, the image 'E12.5 tail' was inadvertently replaced with a duplicate of the image 'E12.5 trunk' from the same panel. In Figure 6d, the image 'E9.5/OH-TAM E8.5, embryo' was inadvertently replaced with a duplicate of the image 'E10.5/ OH-TAM E8.5, embryo' from Fig. 6b. The corrected versions of these figures appear in the Author Correction associated with this Article.

Yolk sac macrophage progenitors traffic to the embryo during defined stages of development.

Tissue macrophages in many adult organs originate from yolk sac (YS) progenitors, which invade the developing embryo and persist by means of local self-renewal. However, the route and characteristics of YS macrophage trafficking during embryogenesis are incompletely understood. Here we show the early migration dynamics of YS-derived macrophage progenitors in vivo using fate mapping and intravital microscopy. From embryonic day 8.5 (E8.5) CX3CR1+ pre-macrophages are present in the mouse YS where they rapidly proliferate and gain access to the bloodstream to migrate towards the embryo. Trafficking of pre-macrophages and their progenitors from the YS to tissues peaks around E10.5, dramatically decreases towards E12.5 and is no longer evident from E14.5 onwards. Thus, YS progenitors use the vascular system during a restricted time window of embryogenesis to invade the growing fetus. These findings close an important gap in our understanding of the development of the innate immune system.

RXRα is upregulated in first trimester endometrial glands of spontaneous abortions unlike LXR and PPARγ.

Nuclear receptors are necessary for uterine invasion of the trophoblast and therefore important for maintaining a viable pregnancy. The aim of this study was to investigate the expression pattern and frequency of LXR, PPARγ and RXRα under physiological circumstances and in spontaneous abortions in endometrial glands and decidual tissue cells. A total of 28 (14 physiologic pregnancies/14 spontaneous abortion) human pregnancies in first trimester were analysed for expression of the nuclear receptors LXR, RXRα and PPARγ. Expression changes were evaluated by immunohistochemistry in decidual tissue and endometrial glands of the decidua. RXRα expression was up-regulated in the endometrial glands of spontaneous abortion (P<0.015). Similar up regulation of RXRα was found in decidual tissue (P<0.05). LXR and PPARγ expression was unchanged in spontaneous abortion. Via Correlation analysis we found a trend to positive correlation of LXR and PPARγ (Spearman correlation coefficient r=0.56 P=0.07) in endometrial glands. In decidual tissue, we found significant negative correlation in the control group, for the combination of RXRα and PPARγ (Spearman correlation coefficient r=0.913, P=0.03). Our data show that RXRα expression is increased in miscarriage in endometrial glands and correlation analysis showed that negative correlation between RXRα and PPARγ disappears in miscarriage. This shift is supposable responsible for the loss of regular function in trophoblast and embryonic tissue.

Galectin-13/PP-13 expression in term placentas of gestational diabetes mellitus pregnancies.

INTRODUCTION: Gestational diabetes mellitus (GDM) is an increasing harm in pregnancy. Inflammatory processes in the placenta seem to have an influence on pathogenesis besides known factors like maternal BMI. Galectin-13 (gal-13) is an immunoregulatory protein, which is suspected to play a role in development of GDM in the placenta. METHODS: A total of 40 placentas were obtained from women treated for gestational diabetes mellitus. Placental tissue for control group was obtained from 40 women with normal pregnancy. We investigated the protein expression of gal-13 in term placentas with immunohistochemistry and immunofluorescence. Immunohistochemical staining was analyzed with the semi-quantified IRS score. Gal-13 serum levels were performed with ELISA on a total of 20 probes from women with GDM and healthy control pregnancies in the third trimester. RESULTS: Gal-13 was found in syncytiotrophoblast, in nuclei of syncytiotrophoblast and trophoblast cells as well in extravillous trophoblast cells of normal placentas. In GDM placentas, gal-13 expression was significantly decreased in all of these examined cell types (syncytiotrophoblast p = 0.003, nuclei of syncytiotrophoblast p = 0.007; extravillous trophoblast cells p = 0.001). The ELISA showed a significant lower gal-13 serum level in blood from pregnant women with GDM in comparison to healthy controls. DISCUSSION: As gal-13 with its anti-inflammatory functions plays a role in regulation of maternal immune system, a lack of gal-13 may contribute to an imbalance in inflammation processes in the placenta during pregnancy and therefore influences development of GDM.

Galectin 2 (gal-2) expression is downregulated on protein and mRNA level in placentas of preeclamptic (PE) patients.

INTRODUCTION: Galectin 2 (gal-2) belongs to the proto type group and consists of two homologous carbohydrate recognition domains (CRDs) resulting in multiple sugar binding sites. The expression of gal-2 has been shown to be involved in processes of angiogenesis and inflammation but was not analyzed before in preeclamptic (PE) placentas. Therefore the aim of this study was an analysis of expression and localization of gal-2 in placentas from patients suffering from PE. METHODS: Placental tissues were obtained from 14 women following a normal course of pregnancy and 13 women with PE. Expression of gal-2 was evaluated with immunohistochemistry and a semi quantitative score. Gal-2 mRNA expression was quantified in placental tissue using real time TaqMan PCR. Identification of gal-2 expressing cells in the decidua was achieved by double immunofluorescence microscopy. RESULTS: Expression of gal-2 is downregulated in the syncytiotrophoblast of preeclamptic placentas. Downregulation of gal-2 could also be identified in the decidua of PE patients. These findings on protein level could be supported by the results of TaqMan PCR. Gal-2 is downregulated in PE placentas on mRNA level. Finally, gal-2 expressing cells could be identified as extravillous trophoblast cells in both normal pregnancy and PE. DISCUSSION: Gal-2 was identified as an inhibitor of arteriogenesis in a murine model supposedly via modulation of the monocyte/macrophage population. In PE, both the formation of spiral arteries as well as influx of macrophages are dramatically changed. Therefore we might speculate that disturbed transformation of spiral arteries in PE might correlate with the downregulated gal-2 expression by the trophoblast.

Poor sleep quality is associated with impaired glucose tolerance in women after gestational diabetes.

We analyzed the association of sleep quality and glucose metabolism in women after gestational diabetes (pGDM) and in women after normoglycemic pregnancy (controls). Data during pregnancy and a visit within the first 15 months after delivery were collected from 61 pGDM and 30 controls in a prospective cohort study. This included a medical history, physical examination, questionnaires (Pittsburgh Sleep Quality Index (PSQI), and Perceived Stress Scale (PSS)), and 5-point oral glucose tolerance test with insulin measurements to determine indices of insulin sensitivity and insulin secretion. We used Spearman correlation coefficients and multivariate regression models for analysis.9.3 ± 3.2 months after delivery, pGDM had significantly higher fasting and 2 h glucose levels and lower insulin sensitivity than controls. There was no significant difference in age, BMI and sleep quality as assessed with the PSQI between the two groups. The PSQI score correlated with the ogtt-2 h plasma glucose in pGDM (δ = 0.41; p = 0.0012), but not in controls. This association was confirmed with a multivariate linear regression model with adjustment for age, BMI and months post-delivery. Perceived stress was an independent risk factor (OR 1.12; 95% CI 1.02-1.23) for impaired sleep. Our findings suggest that post-delivery sleep quality significantly influences glucose tolerance in women after GDM and that impaired sleep is associated with increased stress perception. Measures to improve of sleep quality and reduce perceived stress should therefore be tested as additional strategies to prevent progression to type 2 diabetes after GDM.

Waterbirths: regional audit of infection control practices.

Waterbirths are increasingly being offered as an option, although concerns about associated infection risks have been expressed. We undertook an audit of current practice in our region to help formulate an infection control policy. Questionnaires were sent to the Infection Control Nurses at 16 centres, with a request for completion in conjunction with the Infection Control Doctor. Fourteen (88%) centres responded, 11 of which carried out waterbirthing. Eight had permanent on-site pools, of which five were permanently plumbed in and provided with separate water inlets and outlets. All eight centres had infection control policies for waterbirthing. Six policies included care of the pool using detergent and disinfectant (chlorine-releasing compounds) and two using detergent alone. In none of the centres were the mothers tested for blood-borne viruses. Six centres collected data on infection in mother or child, and two carried out regular microbiological testing of pools. Infection control policies for waterbirths should include instructions for pool maintenance and decontamination, for prevention of legionella, for universal precautions and for use of personal protective equipment. Post-natal surveillance of mothers and babies is required in order to ascertain infection rates.

Detection and characterisation of disseminated tumour cells in bone marrow of breast cancer patients by immunostaining of Her-2 and MUC-1 in combination with Thomsen-Friedenreich (CD176).

Disseminated tumour cells (DTCs) in the bone marrow derive from many primary tumours, such as breast cancer. Their mere existence hints to present or future metastasis and implicates a worse prognosis for the patient. DTCs may possess different characteristics in comparison to the primary tumour due to events like Epithelial-Mesenchymal-Transition. Therefore, these cells might be able to survive chemotherapy and cause relapses of the disease at a later point. We aimed to detect and further characterise DTCs by an immunostaining approach with three different antigen markers (Her-2, MUC-1 and TF, also known as CD 176). For that reason, bone marrow of 41 breast cancer patients was obtained during surgery; DTCs were enriched by density gradient centrifugation and cytospins were prepared. After fixation, immunofluorescent double-stainings were carried out with antibodies against CD176 in combination with HER-2 or MUC-1. Cells co-expressing two antigens were found in all staining combinations (Her-2 and CD176: 46.14%; MUC-1 and CD176: 18.15% of all cases). Cells that stained for a single antigen only were also found (Her-2: 36.86%; MUC-1: 34.45%; CD176: 29.65% of all cases). Significant correlations between the stainings of all markers could be shown (p<0,001). In conclusion, Thomsen-Friedenreich Antigen (TF, CD176) is a promising marker in combination with the established marker Her-2 and other markers like MUC-1. These results may serve as a basis for future DTC detection routines and help to individualize medical treatment, reducing side effects and increasing the efficiency of the therapy.

Expression and function of galectins in the endometrium and at the human feto-maternal interface.

Galectins are classified as lectins that share structural similarities and bind ß-galactosides via a conserved carbohydrate recognition domain. So far 16 out of 19 identified galectins were shown to be present in humans and numerous studies revealed galectins as pivotal modulators of cell death, differentiation and growth. Galectins were highlighted to interact with both the adaptive and innate immune response. In the field of reproductive medicine and placenta research different roles for galectins have been proposed. Several galectins, being abundantly present at the human feto-maternal interphase and endometrium, were hypothesized to significantly contribute to endometrial receptivity and pregnancy physiology. Hence, this review outlines selected aspects of galectin action within endometrial function and at the feto-maternal interphase. Further current knowledge on galectins in reproductive and pregnancy disorders like endometriosis, abortion or preeclampsia is summarized.

[Effects of different methods of tooth resection in suckling piglets]. / Auswirkungen verschiedener Methoden der Zahnresektion beim Saugferkel.

Epizootic examinations were made on 796 newborn piglets to test the consequences of the resection of a piglet's canine and lateral incisor teeth on its first day of life. As a comparative examination the resection was carried out with side-cutter pliers or with the teeth grinder PIGMATIC 110. A third group of piglets was not treated. Histological, radiographic and bacteriological examinations were made of 10 piglets--in each case with piglets with the resection with side-cutter pliers or with the teeth grinder. 48% of ground teeth reacted with an inflammation of the pulp. The clipping of the crown of teeth with side-cutter pliers caused pulpitis in 92% of the teeth. Inflammation of the gingiva could almost only be seen around clipped teeth. Splinters only occurred with teeth which had been clipped with side-cutter pliers. Gingivitis and pulpitis extended along the splinters to the bottom of the root. Bites among the litter mates were more frequent in the group of piglets whose teeth had not been treated than in the group of piglets with resected teeth. Compared to the grinding or the clipping of needle teeth, bites to the sow's udder were much more frequent if the resection had been omitted. During the whole period of examination the mortality of piglets was at its lowest in the group of piglets with clipped teeth. The development of grinding instruments for the resection of needle teeth presents a method which reduces the negative consequences of the conventional resection with side-cutter pliers and shows clearly the advantages of resection. In terms of animal welfare, teeth resection is an amputation and therefore every case requires veterinary justification.