Analysis of the Beta‐Tubulin Gene and Morphological Changes of the Microsporidium Anncaliia algerae both Suggest Albendazole Sensitivity

The Microsporidium, Anncaliia algerae, an obligate intracellular parasite, has been identified as an opportunistic human pathogen, but treatment has not been evaluated for infections with this organism. Albendazole, an antitubulin polymerization drug used against parasitic worm infections, has been the medication of choice used to treat some microsporidial infections affecting humans, with varying results ranging from clearing infection (Encephalitozoon) to resistance (Enterocytozoon). This study illustrates the effect of albendazole treatment on A. algerae infection in Rabbit Kidney (RK13) cells and Human Fetal Lung (HFL‐1) fibroblasts. Albendazole appears to have an attenuating effect on A. algerae infection and albendazole's IC₅₀ in RK13 cells is 0.1 μg/ml. Long‐term treatment inhibits up to 98% of spore production, but interrupting treatment reestablishes the infection without new exposure to the parasite as supported by microscopic observations. The parasite's beta‐tubulin gene was purified, cloned, and sequenced. Five of the six specific amino acids, associated with benzimidazole sensitivity, are conserved in A. algerae. These findings suggest that A. algerae is sensitive to albendazole; however, the organism is not completely cleared from cultures.     

X-ray dark-field phase-contrast imaging: Origins of the concept to practical implementation and applications

The basic idea of X-ray dark-field imaging (XDFI), first presented in 2000, was based on the concepts used in an X-ray interferometer. In this article, we review 20 years of developments in our theoretical understanding, scientific instrumentation, and experimental demonstration of XDFI and its applications to medical imaging. We first describe the concepts underlying XDFI that are responsible for imparting phase contrast information in projection X-ray images. We then review the algorithms that can convert these projection phase images into three-dimensional tomographic slices. Various implementations of computed tomography reconstructions algorithms for XDFI data are discussed. The next four sections describe and illustrate potential applications of XDFI in pathology, musculoskeletal imaging, oncologic imaging, and neuroimaging. The sample applications that are presented illustrate potential use scenarios for XDFI in histopathology and other clinical applications. Finally, the last section presents future perspectives and potential technical developments that can make XDFI an even more powerful tool.     

Host age, sex, and pathogen exposure level as factors in the susceptibility of Musca domestica to Entomophthora muscae

Adult Musca domestica were exposed to conidial showers from cadavers that were killed by Entomophthora muscae. Of the 4 host ages (14 days, 7 days, 3 days and <1 day), older flies were more likely to die without postmortem signs of infection, while younger flies were more likely to die with signs and produce primary conidia. The youngest fly group also succumbed to infection significantly sooner than older flies. Total mortality did not differ for the 2 sexes, but females died significantly more often with signs in one trial. Males died significantly earlier than did females, but differences were not significant when the sexes were paired by weight. Increased levels of pathogen exposure significantly increased both total mortality and the number of flies dying with signs; it also decreased incubation period. The potential significance of these findings in the epidemiology of the disease is discussed.

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Zusammenfassung Die Mortalitätsrate von Musca domestica nach Behandlung mit Entomophthora muscae Sporen wurde in Laborversuchen in Abhängigkeit von Wirtsalter,-geschlecht und Expositionsdauer festgestellt. Fliegen verschiedenen Alters (14 Tage, 7 Tage, 3 Tage und jünger als 1 Tag) wurden gemeinsam in 237 ml Kartonbehältern gehalten. Die Behälter besassen Netzdeckel, auf denen frisch an einer Entomophthora Infektion gestorbene Fliegen ausgelegt wurden. Auf diese Weise wurden die Versuchstiere 8 Stunden (low exposure) bzw. 48 Stunden (high exposure) lang mit Entomophthora Sporen kontaminiert. Mit Hilfe von am Boden der Versuchsgefässe ausgelegten Objektträgern wurde die relative Menge Entomophthora Sporen festgestellt, der die Versuchstiere ausgesetzt waren. Der Totenfall wurde täglich für die Dauer von 10 Tagen registriert; dabei wurde zwischen Konidienträgern (external signs) und Leichen ohne Konidien (no external signs) unterschieden.Die Versuchsgruppen, die 48 Stunden lang mit Entomophthora Sporen behandelt worden waren, wiesen im Vergleich zu den kürzer behandelten Versuchsgruppen höhere Mortalität und einen höheren Anteil an Konidienträgern auf. Darüber hinaus wurde das Auftreten von Konidienträgern bei den länger behandelten Gruppen früher beobachtet. Im Vergleich zu unbehandelten Kontrollgruppen erhöhte die Behandlung mit Entomophthora Sporen auch die Rate nicht konidientragender Leichen.Bei gleicher Expositionsdauer zeigten Weibchen und Männchen ähnliche Mortalitätsraten. Ein höherer Anteil Konidienträger bei den Weibchen wurde bei einem von zwei entsprechenden Versuchen beobachtet. Konidienträger traten bei Weibchen später als bei Männchen auf. In Bezug auf die Inkubationszeit konnte kein geschlechsspezifischer Unterschied festgestellt werden, wenn Versuchsgruppen mit gleichem Gewicht verglichen wurden. Bei beiden Geschlechtern erfolgte die Bildung von Konidien bei grösseren Individuen später als bei kleineren Individuen.Das Alter der Versuchstiere beeinflusste sowohl die Mortalitätsrate als auch die Häufigkeit und das zeitliche Auftreten von Konidienträgern. Jüngere Fliegen wiesen einen höheren Anteil Konidienträger auf; bei Fliegen mit einem Alter von weniger als einem Tag wurde das Auftreten von Konidienträgern früher beobachtet als bei älteren Individuen.Die untersuchten Faktoren können auch für die Epidemiologie von Entomophthora freilebender Musca domestica Populationen eine wichtige Rolle spielen. In Populationen mit niedrigerem Durchschnittsalter wird sich eine Infektion rascher ausbreiten, da jüngere Fliegen eine kürzere Inkubationszeit aufweisen und auch nach kürzerer Kontaminationsdauer Konidien an den getöteten Individuen ausgebildet werden. Darüber hinaus wird sich eine Infektion dann rascher ausbreiten, wenn die Population mit einer grösseren Sporenmenge infiziert wird, da dann eine kürzere Inkubationzeit und ein höherer Anteil Konidienträger zu erwarten ist.

     

Effect of Simulated Acid Rain on Bursaphelenchus xylophilus Infection of Pine Seedlings

White, Scots, and Austrian 3-year-old pine seedlings were treated with conditions simulating acid rain and inoculated with the white pine specific pathotype of Bursaphelenchus xylophilus, VPSt-1. Oleoresin concentration increased slightly and carbohydrate concentration decreased in all seedlings treated with simulated acid rain (SAR). The changes were significantly increased after inoculation of SAR-treated white and Scots pine seedlings with VPSt-1. Wilting was delayed and nematode reproduction decreased in SAR-treated white pine seedlings inoculated with VPSt-1. SAR-treated Austrian pine seedlings were resistant to VPSt-1, but SAR-treated Scots pine seedlings lost tolerance to VPSt-1 and wilted 50-60 days after inoculation.     

Corneal Dystrophy Mutations Drive Pathogenesis by Targeting TGFBIp Stability and Solubility in a Latent Amyloid-forming Domain

Numerous mutations in the corneal protein TGFBIp lead to opaque extracellular deposits and corneal dystrophies (CDs). Here we elucidate the molecular origins underlying TGFBIp's mutation-induced increase in aggregation propensity through comprehensive biophysical and bioinformatic analyses of mutations associated with every major subtype of TGFBIp-linked CDs including lattice corneal dystrophy (LCD) and three subtypes of granular corneal dystrophy (GCD 1–3). LCD mutations at buried positions in the C-terminal Fas1–4 domain lead to decreased stability. GCD variants show biophysical profiles distinct from those of LCD mutations. GCD 1 and 3 mutations reduce solubility rather than stability. Half of the 50 positions within Fas1–4 most sensitive to mutation are associated with at least one known disease-causing mutation, including 10 of the top 11 positions. Thus, TGFBIp aggregation is driven by mutations that despite their physico-chemical diversity target either the stability or solubility of Fas1–4 in predictable ways, suggesting straightforward general therapeutic strategies.     

综述: ILC 细胞的生物学功能与免疫调节作用

ILC细胞即固有淋巴样细胞(innate lymphoid cells),是近年来新发现的一群参与固有免疫的淋巴细胞,根据其表达的转录因子、接受和分泌的细胞因子的不同可以将其分为不同类群．ILC细胞来源于骨髓中的共同淋巴祖细胞(CLP),并可经由淋巴样祖细胞(αLP)、早期淋巴样前体细胞(EILP)、共同辅助淋巴样前体细胞(CHILP)、ILC前体细胞(ILCP)等发育分化为成熟的ILC细胞类群．ILC细胞能够通过与周围环境中的神经细胞、上皮细胞、基质细胞、适应性免疫细胞、髓系细胞、共生菌群等相互作用,协调环境中的信号并广泛参与抗病原体感染、炎症疾病发生、器官形成及组织修复、癌症发生、代谢及生物节律等生物学过程．ILC细胞可能作为新的治疗靶点和治疗手段,对相关疾病进行干预治疗．本文拟就ILC细胞的分类、发育分化、免疫调节、生物学功能、疾病干预策略及研究展望等方面的研究进展进行综述．     

The physiological effects of salmon lice infection on post-smolt of Atlantic salmon

The physiological effects of salmon lice infections on post-smolt of Atlantic salmon were examined by experimentally infecting hatchery reared post-smolts with infective copepodids. Even at high infection intensities, ranging from 30–250 lice per fish, early chalimus stages did not have severe, physiological effects on the fish. There was a sudden increase in fish mortality after the appearance of preadult I stages. Infected fish were then suffering due to lesions and osmoregulatory failure. Plasma chloride level increased significantly and total protein, albumin and haematocrit decreased significantly in infected compared to uninfected fish. All infected fish became moribund before adult lice appeared. Infection intensities above 30 salmon lice larvae per fish thus appear to cause death of Atlantic salmon post-smolt soon after the lice reach their pre-adult stage.     

Cyclone‐based spore trapping,quantitative real‐time polymerase chain reaction and high resolution melting analysis for monitoring airborne inoculum of Magnaporthe oryzae

Rice blast, caused by Magnaporthe oryzae, is one of the most devastating diseases in rice worldwide. We aimed to develop an integrated approach for convenient collection, quantification and characterisation of M. oryzae spores (airborne inoculum) in the field. We developed an easy‐to‐use cyclone‐based spore trap (the AirSampler) and a standard procedure for handling a small amount of airborne spores. Using a specific primer pair or a probe designed for the single‐copy gene mif23, SYBR Green and TaqMan assays could quantify 10 and 4 copy numbers, respectively, of M. oryzae DNA. During 2012 and 2013, the AirSampler and SYBR Green quantitative real‐time polymerase chain reaction were used to monitor temporal dynamics of M. oryzae spores in nursery fields of rice showing symptoms of blast disease. During four cropping seasons, the new techniques could detect M. oryzae spores before the appearance of rice blast symptoms. The amount of spores was low in the early season, then increased, with high fluctuations during the mid‐season and decreased to low levels at the heading stage in the late season. To improve the handling and storage of spore samples, we tested the effect of different treatments on the preservation of spore DNA. DNA loss was reduced with samples protected from ultraviolet B radiation, suspended in CTAB buffer, kept at room temperature or 4°C and used for DNA extraction in 2 weeks. Finally, we demonstrated that the high resolution melting analysis could be used for rapid determination of A, D, A + D and C alleles of the avirulence gene pex31 (Avr‐Pik/kp/km) in M. oryzae.     

Intrauterine trophoblast migration: A comparative view of humans and rodents

ABSTRACT

Trophoblast migration and invasion through the decidua and maternal uterine spiral arteries are crucial events in placentation. During this process, invasive trophoblast replace vascular endothelial cells as the uterine arteries are remodeled to form more permissive vessels that facilitate adequate blood flow to the growing fetus. Placentation failures resulting from either extensive or shallow trophoblastic invasion can cause pregnancy complications such as preeclampsia, intrauterine growth restriction, placenta creta, gestational trophoblastic disease and even maternal or fetal death. Consequently, the use of experimental animal models such as rats and mice has led to great progress in recent years with regards to the identification of mechanisms and factors that control trophoblast migration kinetics. This review aims to perform a comparative analysis of placentation and the mechanisms and factors that coordinate intrauterine trophoblast migration in humans, rats and mice under physiological and pathological conditions.