Detection of Anaplasma phagocytophilum in horses from Germany by molecular and serological testing (2008–2021)

Background Equine granulocytic anaplasmosis (EGA) is a tick-borne disease caused by Anaplasma (A.) phagocytophilum. In Germany, this pathogen is transmitted primarily by Ixodes ricinus. There is limited knowledge about its prevalence in horses in Germany. The aim of this retrospective study was to analyze the results of serological and molecular testing for A. phagocytophilum in horses which were done in a commercial laboratory in Germany over fourteen years. Additionally, risk factors were evaluated, and hematological abnormalities were addressed in horses with positive PCR results. Methods This retrospective study examined results of direct (Polymerase chain reaction [PCR]) and indirect (immunofluorescence antibody test [IFAT]) detection methods for A. phagocytophilum in horses on samples provided by German veterinarians and processed by the commercial laboratory LABOKLIN from 2008 to 2021. In horses with positive test results, a Complete Blood Count (CBC) and Serum Amyloid A (SAA) were also analyzed where possible. Results In total, 1217/4834 horses tested positive (PCR: 190/1246 horses, 15.2%; IFAT: 1036/3849 horses, 26.9%). Seasonality and location, as classified by federal state, had a statistically significant impact on PCR results (P < 0.001 for both). In horses with positive PCR results, hematological abnormalities were detected in 112/118 horses (95%), with thrombocytopenia (86%) and anemia (52%) representing the most common findings. The remaining 6/118 horses (5%) showed no hematological abnormalities on CBC. SAA was measured in 35 horses with positive PCR results, which exclusively showed marked elevation. Conclusions The seasonality of A. phagocytophilum infections confirmed by PCR testing was consistent with known peaks in vector activity in Germany. The high rate of horses with positive PCR results when compared to dogs and cats may be due to a lack of ectoparasite prophylaxis. Infections with A. phagocytophilum should be considered as a differential diagnosis in horses with cytopenia on CBC and SAA elevation, especially in the summer and after any possible tick exposure

Molecular and Serological Detection of Anaplasma phagocytophilum in Dogs from Germany (2008–2020)

Anaplasma phagocytophilum is an obligate intracellular bacterium that causes granulocytic anaplasmosis in domestic animals, wildlife, and humans and is primarily transmitted by ticks of the Ixodes persulcatus complex. This retrospective study aims to determine the percentages of dogs that tested positive for A. phagocytophilum in Germany. It included the results of direct (polymerase chain reaction [PCR]) and indirect (immunofluorescence antibody test [IFAT], antibody-enzyme-linked immunosorbent assay [ELISA]) detection methods performed in the laboratory LABOKLIN on canine samples provided by German veterinarians from 2008 to 2020. Out of a total of 27,368 dogs tested by PCR, 1332 (4.9%) tested positive, while 24,720 (27.4%) of the 90,376 dogs tested by IFAT/ELISA had positive serology. High rates of positive PCR results were observed in months with known peaks in vector activity, showing that the dynamics of A. phagocytophilum infections in dogs in Germany are consistent with vector activity. In dogs with a positive PCR result, peaks in serology could be observed four weeks after initial testing. Male and senior dogs had higher rates of positive serology. A possible impact of environmental factors such as changes in climate should be investigated further. Overall, the upward trend in positive test results over the years indicates that canine granulocytic anaplasmosis will continue to become increasingly important for veterinary medicine

Ethics and accuracy in scientific researches with emphasize on taxonomic works

Research is one of those highly complex and subtle activities that usually remain quite unformulated in the minds of those who practise them. This is probably why most scientists think that it is not possible to give nay formal instruction in how to do research. Additionally, training in research must be largely self-training,preferably with the guidance of an experienced scientist in the handling of the actual investigation. Scientific research, which is simply the search for new knowledge, appeals especially to people who are individualists and their method vary from one person to another. A policy followed by one scientist may not be suitable for another, and different methods are required in different branches of science. However, there are some basic principles and mental techniques that are commonly used in most types of investigation, at least in the biological sphere. Honesty and morals are the first step in all scientific researches and a researcher must consider it in all stages of a scientific work. The role and importance of accuracy and honesty in taxonomy and systematic is more than other biological sciences. For having the honesty in scientific researches, learning the principals of ethics is needed for all the researchers and students. Therefore, this paper deals with the ethics in science and technology and also important comments on taxonomy.Facultad de Ciencias Naturales y Muse

Ethics and accuracy in scientific researches with emphasize on taxonomic works

Research is one of those highly complex and subtle activities that usually remain quite unformulated in the minds of those who practise them. This is probably why most scientists think that it is not possible to give nay formal instruction in how to do research. Additionally, training in research must be largely self-training,preferably with the guidance of an experienced scientist in the handling of the actual investigation. Scientific research, which is simply the search for new knowledge, appeals especially to people who are individualists and their method vary from one person to another. A policy followed by one scientist may not be suitable for another, and different methods are required in different branches of science. However, there are some basic principles and mental techniques that are commonly used in most types of investigation, at least in the biological sphere. Honesty and morals are the first step in all scientific researches and a researcher must consider it in all stages of a scientific work. The role and importance of accuracy and honesty in taxonomy and systematic is more than other biological sciences. For having the honesty in scientific researches, learning the principals of ethics is needed for all the researchers and students. Therefore, this paper deals with the ethics in science and technology and also important comments on taxonomy.Facultad de Ciencias Naturales y Muse

Random Fan-Out State Induced by Site-Random Interlayer Couplings

We study the low-temperature properties of a classical Heisenberg model with site-random interlayer couplings on the cubic lattice. This model is introduced as a simplified effective model of Sr(Fe$_{1-x}$Mn$_{x}$)O$_2$, which was recently synthesized. In this material, when $x=0.3$, $(\pi\pi\pi)$ and $(\pi\pi0)$ mixed ordering is observed by neutron diffraction measurements. By Monte Carlo simulations, we find an exotic bulk spin structure that explains the experimentally obtained results. We name this spin structure the "random fan-out state". The mean-field calculations provide an intuitive understanding of this phase being induced by the site-random interlayer couplings. Since Rietveld analysis assuming the random fan-out state agrees well with the neutron diffraction pattern of Sr(Fe$_{0.7}$Mn$_{0.3}$)O$_2$, we conclude that the random fan-out state is reasonable for the spin-ordering pattern of Sr(Fe$_{0.7}$Mn$_{0.3}$)O$_2$ at the low-temperature phase.Comment: 13 pages, 12 figure

Citrobacter rodentium Subverts ATP Flux and Cholesterol Homeostasis in Intestinal Epithelial Cells In Vivo.

The intestinal epithelial cells (IECs) that line the gut form a robust line of defense against ingested pathogens. We investigated the impact of infection with the enteric pathogen Citrobacter rodentium on mouse IEC metabolism using global proteomic and targeted metabolomics and lipidomics. The major signatures of the infection were upregulation of the sugar transporter Sglt4, aerobic glycolysis, and production of phosphocreatine, which mobilizes cytosolic energy. In contrast, biogenesis of mitochondrial cardiolipins, essential for ATP production, was inhibited, which coincided with increased levels of mucosal O2 and a reduction in colon-associated anaerobic commensals. In addition, IECs responded to infection by activating Srebp2 and the cholesterol biosynthetic pathway. Unexpectedly, infected IECs also upregulated the cholesterol efflux proteins AbcA1, AbcG8, and ApoA1, resulting in higher levels of fecal cholesterol and a bloom of Proteobacteria. These results suggest that C. rodentium manipulates host metabolism to evade innate immune responses and establish a favorable gut ecosystem

Cyclosporin A Treatment of Leishmania donovani Reveals Stage-Specific Functions of Cyclophilins in Parasite Proliferation and Viability

BACKGROUND: Cyclosporin A (CsA) has important anti-microbial activity against parasites of the genus Leishmania, suggesting CsA-binding cyclophilins (CyPs) as potential drug targets. However, no information is available on the genetic diversity of this important protein family, and the mechanisms underlying the cytotoxic effects of CsA on intracellular amastigotes are only poorly understood. Here, we performed a first genome-wide analysis of Leishmania CyPs and investigated the effects of CsA on host-free L. donovani amastigotes in order to elucidate the relevance of these parasite proteins for drug development. METHODOLOGY/PRINCIPAL FINDINGS: Multiple sequence alignment and cluster analysis identified 17 Leishmania CyPs with significant sequence differences to human CyPs, but with highly conserved functional residues implicated in PPIase function and CsA binding. CsA treatment of promastigotes resulted in a dose-dependent inhibition of cell growth with an IC50 between 15 and 20 µM as demonstrated by proliferation assay and cell cycle analysis. Scanning electron microscopy revealed striking morphological changes in CsA treated promastigotes reminiscent to developing amastigotes, suggesting a role for parasite CyPs in Leishmania differentiation. In contrast to promastigotes, CsA was highly toxic to amastigotes with an IC50 between 5 and 10 µM, revealing for the first time a direct lethal effect of CsA on the pathogenic mammalian stage linked to parasite thermotolerance, independent from host CyPs. Structural modeling, enrichment of CsA-binding proteins from parasite extracts by FPLC, and PPIase activity assays revealed direct interaction of the inhibitor with LmaCyP40, a bifunctional cyclophilin with potential co-chaperone function. CONCLUSIONS/SIGNIFICANCE: The evolutionary expansion of the Leishmania CyP protein family and the toxicity of CsA on host-free amastigotes suggest important roles of PPIases in parasite biology and implicate Leishmania CyPs in key processes relevant for parasite proliferation and viability. The requirement of Leishmania CyP functions for intracellular parasite survival and their substantial divergence form host CyPs defines these proteins as prime drug targets

Prediction of hot spot residues at protein-protein interfaces by combining machine learning and energy-based methods

Background: Alanine scanning mutagenesis is a powerful experimental methodology for investigating the structural and energetic characteristics of protein complexes. Individual aminoacids are systematically mutated to alanine and changes in free energy of binding (Delta Delta G) measured. Several experiments have shown that protein-protein interactions are critically dependent on just a few residues ("hot spots") at the interface. Hot spots make a dominant contribution to the free energy of binding and if mutated they can disrupt the interaction. As mutagenesis studies require significant experimental efforts, there is a need for accurate and reliable computational methods. Such methods would also add to our understanding of the determinants of affinity and specificity in protein-protein recognition.Results: We present a novel computational strategy to identify hot spot residues, given the structure of a complex. We consider the basic energetic terms that contribute to hot spot interactions, i.e. van der Waals potentials, solvation energy, hydrogen bonds and Coulomb electrostatics. We treat them as input features and use machine learning algorithms such as Support Vector Machines and Gaussian Processes to optimally combine and integrate them, based on a set of training examples of alanine mutations. We show that our approach is effective in predicting hot spots and it compares favourably to other available methods. In particular we find the best performances using Transductive Support Vector Machines, a semi-supervised learning scheme. When hot spots are defined as those residues for which Delta Delta G >= 2 kcal/mol, our method achieves a precision and a recall respectively of 56% and 65%.Conclusion: We have developed an hybrid scheme in which energy terms are used as input features of machine learning models. This strategy combines the strengths of machine learning and energy-based methods. Although so far these two types of approaches have mainly been applied separately to biomolecular problems, the results of our investigation indicate that there are substantial benefits to be gained by their integration

Severe malaria in children leads to a significant impairment of transitory otoacoustic emissions--a prospective multicenter cohort study.

BACKGROUND: Severe malaria may influence inner ear function, although this possibility has not been examined prospectively. In a retrospective analysis, hearing impairment was found in 9 of 23 patients with cerebral malaria. An objective method to quickly evaluate the function of the inner ear are the otoacoustic emissions. Negative transient otoacoustic emissions are associated with a threshold shift of 20 dB and above. METHODS: This prospective multicenter study analyses otoacoustic emissions in patients with severe malaria up to the age of 10 years. In three study sites (Ghana, Gabon, Kenya) 144 patients with severe malaria and 108 control children were included. All malaria patients were treated with parental artesunate. RESULTS: In the control group, 92.6 % (n = 108, 95 % confidence interval 86.19-6.2 %) passed otoacoustic emission screening. In malaria patients, 58.5 % (n = 94, malaria vs controls p < 0.001, 95 % confidence interval 48.4-67.9 %) passed otoacoustic emission screening at the baseline measurement. The value increased to 65.2 % (n = 66, p < 0.001, 95 % confidence interval 53.1-75.5 %) at follow up 14-28 days after diagnosis of malaria. The study population was divided into severe non-cerebral malaria and severe malaria with neurological symptoms (cerebral malaria). Whereas otoacoustic emissions in severe malaria improved to a passing percentage of 72.9 % (n = 48, 95 % confidence interval 59-83.4 %) at follow-up, the patients with cerebral malaria showed a drop in the passing percentage to 33 % (n = 18) 3-7 days after diagnosis. This shows a significant impairment in the cerebral malaria group (p = 0.012 at days 3-7, 95 % confidence interval 16.3-56.3 %; p = 0.031 at day 14-28, 95 % confidence interval 24.5-66.3 %). CONCLUSION: The presented data show that 40 % of children have involvement of the inner ear early in severe malaria. In children, audiological screening after severe malaria infection is not currently recommended, but is worth investigating in larger studies

A Mechanism for the Polarity Formation of Chemoreceptors at the Growth Cone Membrane for Gradient Amplification during Directional Sensing

Accurate response to external directional signals is essential for many physiological functions such as chemotaxis or axonal guidance. It relies on the detection and amplification of gradients of chemical cues, which, in eukaryotic cells, involves the asymmetric relocalization of signaling molecules. How molecular events coordinate to induce a polarity at the cell level remains however poorly understood, particularly for nerve chemotaxis. Here, we propose a model, inspired by single-molecule experiments, for the membrane dynamics of GABA chemoreceptors in nerve growth cones (GCs) during directional sensing. In our model, transient interactions between the receptors and the microtubules, coupled to GABA-induced signaling, provide a positive-feedback loop that leads to redistribution of the receptors towards the gradient source. Using numerical simulations with parameters derived from experiments, we find that the kinetics of polarization and the steady-state polarized distribution of GABA receptors are in remarkable agreement with experimental observations. Furthermore, we make predictions on the properties of the GC seen as a sensing, amplification and filtering module. In particular, the growth cone acts as a low-pass filter with a time constant ∼10 minutes determined by the Brownian diffusion of chemoreceptors in the membrane. This filtering makes the gradient amplification resistent to rapid fluctuations of the external signals, a beneficial feature to enhance the accuracy of neuronal wiring. Since the model is based on minimal assumptions on the receptor/cytoskeleton interactions, its validity extends to polarity formation beyond the case of GABA gradient sensing. Altogether, it constitutes an original positive-feedback mechanism by which cells can dynamically adapt their internal organization to external signals