Retrospective study on the distribution of Dirofilaria immitis in dogs in Hungary.

Europe has experienced the spreading of vector-borne helminths including heartworms (Dirofilaria immitis) from the Mediterranean countries towards the northern ones in the past decades. Recently, the establishment of D. immitis was confirmed in Hungary on the basis of period prevalence studies involving dogs, red foxes (Vulpes vulpes) and golden jackals (Canis aureus). The aim of our retrospective study was to describe the spatial distribution of the parasite and the time course of spreading of D. immitis in Hungary. Necropsy records of 2622 dogs received at our laboratories from 2001 to 2015 were reviewed for heartworm infections. The locality of origin of animals was recorded in a geographic information system database and compared to the results of the period prevalence study involving wild canids. Autochthonous heartworm infection was detected in 27 dogs. The time course analysis indicates that the parasite established in Hungary in 2007. As temperature is one of the most important determinants of the distribution of D. immitis, the climate of the Great Hungarian Plain is the most suitable region for the establishment of D. immitis in Hungary. Our studies revealed that the Great Hungarian Plain became a D. immitis endemic region for 2015. Nevertheless, sporadic cases in wild canids and dogs also occur in other regions of the country.

Generation of transgene-free mouse induced pluripotent stem cells using an excisable lentiviral system.

One goal of research using induced pluripotent stem cell (iPSC) is to generate patient-specific cells which can be used to obtain multiple types of differentiated cells as disease models. Minimally or non-integrating methods to deliver the reprogramming genes are considered to be the best but they may be inefficient. Lentiviral delivery is currently among the most efficient methods but it integrates transgenes into the genome, which may affect the behavior of the iPSC if integration occurs into an important locus. Here we designed a polycistronic lentiviral construct containing four pluripotency genes with an EGFP selection marker. The cassette was excisable with the Cre-loxP system making possible the removal of the integrated transgenes from the genome. Mouse embryonic fibroblasts were reprogrammed using this viral system, rapidly resulting in large number of iPSC colonies. Based on the lowest EGFP expression level, one parental line was chosen for excision. Introduction of the Cre recombinase resulted in transgene-free iPSC subclones. The effect of the transgenes was assessed by comparing the parental iPSC with two of its transgene-free subclones. Both excised and non-excised iPSCs expressed standard pluripotency markers. The subclones obtained after Cre recombination were capable of differentiation in vitro, in contrast to the parental, non-excised cells and formed germ-line competent chimeras in vivo.

Generation of rabbit pluripotent stem cell lines.

Pluripotent stem cells have the capacity to divide indefinitely and to differentiate into all somatic cells and tissue lines. They can be genetically manipulated in vitro by knocking genes in or out, and therefore serve as an excellent tool for gene function studies and for the generation of models for some human diseases. Since 1981, when the first mouse embryonic stem cell (ESC) line was generated, many attempts have been made to generate pluripotent stem cell lines from other species. Comparative characterization of ESCs from different species would help us to understand differences and similarities in the signaling pathways involved in the maintenance of pluripotency and the initiation of differentiation, and would reveal whether the fundamental mechanism controlling self-renewal of pluripotent cells is conserved across different species. This report gives an overview of research into embryonic and induced pluripotent stem cells in the rabbit, an important nonrodent species with considerable merits as an animal model for specific diseases. A number of putative rabbit ESC and induced pluripotent stem cell lines have been described. All of them expressed stem cell-associated markers and maintained apparent pluripotency during multiple passages in vitro, but none have been convincingly proven to be fully pluripotent in vivo. Moreover, as in other domestic species, the markers currently used to characterize the putative rabbit ESCs are suboptimal because recent studies have revealed that they are not always specific to the pluripotent inner cell mass. Future validation of rabbit pluripotent stem cells would benefit greatly from a validated panel of molecular markers specific to pluripotent cells of the developing rabbit embryos. Using rabbit-specific pluripotency genes may improve the efficiency of somatic cell reprogramming for generating induced pluripotent stem cells and thereby overcome some of the challenges limiting the potential of this technology.

Experimental study on the role of Brachyspira alvinipulli in intestinal spirochaetosis of geese.

Ten one-day-old goslings were inoculated orally with a Brachyspira alvinipulli strain isolated from the large intestine of geese that had died of intestinal spirochaetosis (Group A), 10 day-old goslings were inoculated orally with a B. hyodysenteriae strain (Group B), and a third group of 10 goslings (Group C) served as uninfected control. The goslings were observed daily for clinical signs. They were sacrificed on days 7, 14, 21 and 35 days postinfection (PI), and necropsied. Segments of the large intestine were subjected to histopathological, immunohistochemical, electron microscopic (TEM, SEM) and microbiological examinations. Mortality did not occur during the experimental period. However, in both groups the caecum of the goslings killed by bleeding was slightly dilated, in its lumen there was a watery, yellowish and frothy content, and the mucous membrane was slightly swollen. By histopathological, immunohistochemical and electron microscopic examination, B. alvinipulli and B. hyodysenteriae could be detected in the caecum or colon, in the lumen of the glands and sometimes among the glandular epithelial cells in goslings of the respective groups, and could be reisolated from these organs by culturing. A mild inflammation of the intestinal mucosa was also noted. In transverse section of the brachyspirae, numerous (16-22) periplasmic flagella could be detected inside the outer sheath, also depending on the plane of section.

Typhlocolitis associated with spirochaetes in goose flocks.

The role of Brachyspira bacteria in the aetiology of increased mortality observed in two breeder goose flocks (Flock A consisting of 1,500 and Flock B comprising 4,500 laying geese) at the end of the first egg-laying season, in the period of moulting, was studied. In Flock A 415 geese (28%) died during an 8-week period while in Flock B 834 geese (18%) died during a 12-week period. On gross pathological examination, the geese were found to have haemorrhagic-to-necrotic inflammation of the large intestine (colon and rectum) and fibrinonecrotic typhlitis accompanied by severe degeneration. Often, fibrosis of the kidneys, and in five of the geese secondary visceral urate deposition ("visceral gout") was also observed. Histopathological examination consistently demonstrated spirochaetes in the mucous membrane of the affected large intestine. This was confirmed by the results of immunohistochemical and electron microscopic examination. In addition, Trichomonas stages were also detected from the large intestine of 11 geese. On the basis of their cultural and biochemical properties, and PCR sequencing analysis, eight out of the nine spirochaete strains isolated from the geese by culture on special media under anaerobic conditions were identified as Brachyspira alvinipulli. This is the first report on the isolation of B. alvinipulli from laying geese affected with fibrinonecrotic typhlocolitis.

Prediction of non-sentinel lymph node status in breast cancer with a micrometastatic sentinel node.

BACKGROUND: Axillary lymph node dissection (ALND) may not be necessary in women with breast cancer who have micrometastasis in a sentinel node (SN), owing to the low risk of non-SN (NSN) involvement. The aim of this study was to identify a subgroup of women with a micrometastatic SN and a negligible risk of positive NSNs in whom ALND may be avoided. METHODS: Some 237 of 241 women with a macrometastatic SN and 122 of 138 with a micrometastatic SN underwent completion ALND and were compared with respect to NSN involvement. The 122 patients with SN micrometastasis were further analysed to determine factors that could predict the risk of positive NSNs. RESULTS: A total of 121 (51.1 per cent) of 237 women with SN macrometastasis had positive NSNs compared with 22 (18.0 per cent) of 122 with SN micrometastasis (P < 0.001). Multivariate analysis showed that size of SN micrometastasis (odds ratio 3.49 (95 per cent confidence interval (c.i.) 1.32 to 9.23); P = 0.012) and presence of lymphovascular invasion (odds ratio 0.23 (95 per cent c.i. 0.05 to 1.00); P = 0.050) were significantly associated with positive NSNs. SN micrometastasis less than 0.5 mm in diameter combined with absence of lymphovascular invasion was associated with an 8.5 per cent risk of NSN involvement. CONCLUSION: Size of micrometastasis and presence of lymphovascular invasion were significantly related to the risk of finding additional positive axillary lymph nodes when the SN contained only micrometastasis.

Direct measurement of the viscoelasticity of adsorbed protein layers using atomic force microscopy.

Thick layers of the protein lysozyme have been deposited on mica, and their force-distance hysteresis measured using atomic force microscopy in the presence of different salts. Sodium thiocyanate, which is known to lower the melting temperature of proteins and increase their solubility, increases lysozyme deformability and lowers the viscosity of the protein layer, compared with sodium chloride. Sodium phosphate, known to raise the melting temperature and lower the solubility, decreases deformability and increases the viscosity.

[Aloys Martin and the Society of German Physicians in Paris]. / Aloys Martin und der Verein deutscher Aerzte in Paris.

The news about the discovery of the anaesthetic properties of ether swept rapidly from America to Europe. The first informations concerning this topic reached England in late fall of 1846 and after that came to France and especially Paris. The German physician Aloys Martin from Munich who was living in Paris at that time was largely responsible for conveying the news about these new findings in medical science to the German-speaking nations. Controversial discussions concerning the anaesthetic properties of ether developed among the members of various scientific circles right after the news from America had reached the city--and many members of the "Verein Deutscher Aerzte in Paris" (Association of German Physicians in Paris) took part in them, Martin then was one of the club's most active members and repeatedly reported on the new method as a correspondent for the widely read "Augsburger Allgemeine Zeitung": Documents discovered recently at the archives of the "Leopoldinisch-Carolinischen Akademie" of Halle provide an interesting insight into this association, whose members tried to find out more about the anaesthetic properties and side effects of ether carrying out a series of well appreciated self-experiments in the spring of 1847. The following text reports on the history of the "Verein Deutscher Aerzte in Paris" which has largely fallen into oblivion and emphasizes the importance of Aloys Martin whose name is strongly attached to the spread of the use of ether throughout the German-speaking countries.

[Heparin-induced changes in blood gas analysis (author's transl)]. / Heparinbedingte Veränderungen der Blutgaswerte.

In modern blood gas analysis heparin is used for preventing coagulation in the syringe and the analyzer. If the amount of heparin added is too large the pH of the blood sample falls and so an artificial metabolic acidosis is induced. Consequently the amount of heparin used should be just enough to prevent blood coagulation without changing appreciably the acid-base status of the sample. High concentrations of inspired oxygen have no additional effect on the ability of heparin to alter the pH values.

[A simple universally applicable I.M.V. system (author's transl)]. / Ein einfaches, universell verwendbares I.M.V.-System.

A simple I.M.V.-system is described based on the AMBU-E-valve applied to pressure-cycled ventilators (e.g. Bird Mark VII).

[A simple system for the removal of excess anaesthetic vapours for use with the Jackson-Rees modification of Ayres' T-Piece (author's transl)]. / Einfaches System zur Absaugung von Narkosegasen bei Verwendung der Jackson-Rees-Modifikation von Ayres T-Stück

Details are given of the arrangement and functioning of a simple anti-pollution system for use with anaesthetic apparatus without re-breathing.