Setaria tundra in a roe deer (Capreolus capreolus) in the Donau-Ries district of Bavaria, Germany.

INTRODUCTION: Setaria tundra is known as a common parasite of sylvatic ungulates in Northern latitudes. Although mostly considered of low pathogenicity, severe disease outbreaks and remarkable economic losses have been observed in reindeer (Rangifer tarandus tarandus). Host density and climatic factors are major drivers of the expansion of Setaria spp. facilitating their development and spread. Five adult specimens of S. tundra were retrieved from a male roe deer in Bavaria, Germany. Deoxyribonucleic acid (DNA) barcoding confirmed morphological identification. Cyclooxygenase 1 gene sequences showed 98,73-99,68 % similarity to sequences of other S. tundra specimens found in deer (Cervidae) and mosquitoes (Culicidae). The results raise awareness for the presence of S. tundra in a hitherto unkown endemic region and represent a starting point for broader investigations to understand the biology and distribution of this parasite in roe deer as well as other potential definitive hosts.

INTRODUCTION: Setaria toundra est connu comme un parasite commun des ongulés sylvatiques aux latitudes septentrionales. Bien que généralement considéré comme faiblement pathogène, de graves épidémies et des pertes économiques significatives ont été observées chez le renne (Rangifer tarandus tarandus). La densité d'hôtes et les facteurs climatiques sont les principaux moteurs de l'expansion de Setaria spp., facilitant leur développement et leur diffusion. Cinq spécimens adultes de S. toundra ont été récupérés sur un chevreuil mâle en Bavière, en Allemagne. Le séquençage de l'acide désoxyribonucléique (ADN) a confirmé l'identification morphologique. Les séquences du gène de la cyclooxygénase 1 ont montré une similarité de 98,73 à 99,68 % avec les séquences d'autres spécimens de S. toundra trouvés chez les cerfs (Cervidae) et les moustiques (Culicidae). Les résultats sensibilisent à la présence de S. toundra dans une région endémique jusqu'alors inconnue et représentent un point de départ pour des investigations plus larges pour comprendre la biologie et la distribution de ce parasite chez le chevreuil ainsi que d'autres hôtes définitifs potentiels.

Novel MLST sequence types of pathogenic Leptospira spp.: Opening the black box of animal leptospirosis in Brazil.

In the current context of the emergence of certain infectious diseases and discussion of the One Health concept for many of these, the study of leptospirosis - both in domestic and wild hosts - cannot be neglected. The study of animal leptospirosis has evolved in recent years. It has been demonstrated that the human-animal-environment interface is more important than previously thought. In the present study, 35 strains of five pathogenic Leptospira species were isolated from different animal species in Brazil and characterized by rrs, secY, and Multilocus Sequence Typing (MLST) sequencing. Phylogenetic inferences were performed and the molecular diversity of the populations (intra- and inter-population levels) was evaluated. Among the five studied species, 18 different sequence types (STs) were found (22 new alleles and 11 new STs). eBURST analysis revealed two clonal complexes (CCs) and seven singletons. A high genetic diversity was demonstrated (H = 0.954 ± 0.017), mainly for the L. santarosai population (H = 0.942 ± 0.034, n = 20). The same strain was identified in different host species, as well as strains with zoonotic potential circulating in the country. Although the difficulty of culturing Leptospira strains is well known, the high variability of the strains found in Brazil highlights the importance of animals in maintaining the biological cycle of the bacterium in nature. Moreover, the selection of autochthonous strains for the development of vaccines becomes a challenge.

Kato-Katz thick smears as a DNA source of soil-transmitted helminths.

Despite the reduction in the prevalence of soil-transmitted helminthiases in many regions of the world, morbidity rates remain high in some rural regions. The Kato-Katz technique is a simple, inexpensive and field-applicable tool commonly used for the diagnosis and worm-burden characterization of these infections. Molecular studies have revolutionized our understanding of the epidemiology and evolutionary genetics of parasites. In this study we recovered helminthic DNA from Kato-Katz slides (n = 93) prepared in 2011 in the Brazilian Amazon. We achieved DNA recovery by polymerase chain reaction (PCR) in 84% of cases for Ascaris sp. and 75% of cases for hookworms. The sequencing confirmed the specific species of the amplicons. The slides stored for a few years could be analysed using this methodology, allowing access to DNA from a large collection of samples. We must consider the Kato-Katz thick smears as a source of helminth DNA. This can significantly reduce logistical difficulties in the field in terms of obtaining, preserving, transporting and initial processing of samples.

VNTR analysis demonstrates new patterns and high genetic diversity of Leptospira sp. of animal origin in Brazil.

Leptospirosis is a worldwide zoonosis with a broad host range, including humans and domestic and wild animals. The taxonomic classification of Leptospira species is complex. More recently, several molecular tools have been employed in the attempt to group the different strains of leptospires. The objective of this study was the genetic characterization of Brazilian Leptospira sp. isolates obtained from wild and domestic animals. The genotyping of the strains was performed by the variable-number tandem repeats (VNTR) technique, using the Leptospira interrogans/Leptospira kirschneri and Leptospira santarosai protocols. A total of 27 novel strains were characterized and 22 novel VNTR patterns were described. This study suggests a high genetic diversity among the strains obtained from different wild and domestic animals, and reinforces the need for a broad approach aligned to the One Health concept for leptospirosis. SIGNIFICANCE AND IMPACT OF THE STUDY: Animal leptospirosis is an important cause of reproductive failure in livestock and economic losses to producing countries. This study describes several novel variable-number tandem repeats patterns and points out the high genetic diversity of Brazilian strains. Understanding the circulation of strains between animals is essential for the control of leptospirosis in livestock.

Gene silencing of stearoyl-ACP desaturase enhances the stearic acid content in Chlamydomonas reinhardtii.

In this study, stearoyl-ACP desaturase (SAD), the enzyme that converts stearic acid into oleic acid, is silenced by artificial microRNA in the green microalga Chlamydomonas reinhardtii. Two different constructs, which target different positions on the mRNA of stearoyl-ACP desaturase, were tested. The mRNA levels for SAD were reduced after the silencing construct was induced. In one of the strains, the reduction in SAD mRNA resulted in a doubling of the stearic acid content in triacylglycerol molecules, which shows that stearic acid production in microalgae is possible.

Draft Genome Sequence of the Oleaginous Green Alga Tetradesmus obliquus UTEX 393.

The microalgae Tetradesmus obliquus is able to maintain a high photosynthetic efficiency under nitrogen limitation and is considered a promising green microalgae for sustainable production of diverse compounds, including biofuels. Here, we report the first draft whole-genome shotgun sequencing of T. obliquus The final assembly comprises 108,715,903 bp with over 1,368 scaffolds.

[Patients' rights--doctors' duties]. / Patientenrechte--Arztpflichten (I).

On 26 February 2013 the new "Law on Patients' Rights" (hereinafter also the "Law") became effective. This Law strengthens patients' rights vis-à-vis the insurdnce company and also regulates patients' rights regarding their relation to the doctor. This has consequences for the laws on medical liability all doctors must consider. The doctor's performance is and remains a service and such service does not hold any guarantee of success. Nevertheless, this Law primarily reads as a "law on the duties of physicians". To duly take into account these duties and to avoid mistakes and misinterpretation of the Law, the Ethics Committee of the Consortium of Osteosynthesis Trauma Germany (AOTRAUMA-D) has drafted comments on the Law. Brief summaries of its effects are to be found at the end of the respective comment under the heading "Consequences for Practice". The text of the law was influenced particularly by case law, as continuously developed by the German Federal Court of Justice ("BGH"). The implementation of the Law on Patients' Rights was effected by the newly inserted sections 630a to 630h of the German Civil Code (the "BGB"), which are analysed below. The following comments are addressed to physicians only and do not deal with the specific requirements and particularities of the other medical professions such as physiotherapy, midwifery and others so on. Special attention should be paid to the comments on the newly inserted Duty to inform, which has to be fullfilled prior to any diagnostic or therapeutic procedure (sec. 630c para 2 sentence 1 BGB). Under certain conditions the doctor also has to inform the patient about the circumstances that lead to the presumed occurance of a therapeutic or diagnostic malpractice (sec. 630c para. 2 sentence 2 BGB), based on the manifestation of an undesired event or an undesired outcome. As before, the patient's valid consent to any procedure (sec. 630d BGB) is directly linked to the comprehensive and timely provision of information (sec. 630e BGB). Comprehensive documentation obligations regarding all procedures are stipulated in sec. 630f BGB. As before, the burden of proof still rests with the patient, unless a severe malpractice has been established (sec. 630h BGB). The definition of "severe malpractice" remains unchanged and is based on the case law of the Federal Court of Justice (BGH). The patient's obligations to preserve his or her health and to actively support the process of recovery and securing a positive outcome of the treatment are not explicitly mentioned in the Law. Nevertheless, the patient and the physician need to work closely together to achieve a successful result of the treatment. In case the patient does not give his or her cooperation, the physician should consider terminating the treatment relationship.

Prevalence and epidemiology of intestinal parasitism, as revealed by three distinct techniques in an endemic area in the Brazilian Amazon.

This survey aims to estimate the prevalence of intestinal parasitic infections in Santa Isabel do Rio Negro, Amazonian Brazil, through three distinct techniques, correlating the prevalence rates with family income and age groups as well as assessing the household clustering of infections. Prevalence rates were assessed through Graham (n=113), Baermann-Moraes (n=232) and Ritchie (n=463) methods. The Graham method was adopted only for children under 5 years old, 15% of whom were positive for Enterobius vermicularis. By the Baermann-Moraes technique, 5·6% of the samples were positive for Strongyloides stercoralis larvae. The Ritchie technique disclosed the following results: Ascaris lumbricoides (26%), Trichuris trichiura (22·5%), hookworms (9·5%), Entamoeba histolytica/Entamoeba dispar (25·3%), Giardia lamblia (12·5%) and E. vermicularis (0·6%). Children aged 5-14 years presented the highest prevalence for pathogenic parasites. Giardiasis and hookworm infection rates were inversely related to family income. The presence of positive contacts in the same household substantially increased the risk of infection by enteric parasites: odds ratio (OR)=2·70, 95% confidence interval (CI)=1·69-4·29 for ascariasis; OR=2·17, 95% CI=1·34-3·51 for trichuriasis; OR=2·13, 95% CI=1·08-4·17 for hookworm disease; OR=3·42, 95% CI=1·86-6·30 for giardiasis; and OR=2·16, 95% CI=1·35-3·47 for amoebiasis, supporting infection clustering in the home. Intestinal parasitoses are extremely frequent in the studied area, and routine methods for diagnosis may underestimate the prevalence of enterobiasis and strongyloidiasis.

Top-down causation by information control: from a philosophical problem to a scientific research programme.

It has been claimed that different types of causes must be considered in biological systems, including top-down as well as same-level and bottom-up causation, thus enabling the top levels to be causally efficacious in their own right. To clarify this issue, the important distinctions between information and signs are introduced here and the concepts of information control and functional equivalence classes in those systems are rigorously defined and used to characterize when top-down causation by feedback control happens, in a way that is testable. The causally significant elements we consider are equivalence classes of lower level processes, realized in biological systems through different operations having the same outcome within the context of information control and networks.

Large phenotype jumps in biomolecular evolution.

By defining the phenotype of a biopolymer by its active three-dimensional shape, and its genotype by its primary sequence, we propose a model that predicts and characterizes the statistical distribution of a population of biopolymers with a specific phenotype that originated from a given genotypic sequence by a single mutational event. Depending on the ratio g(0) that characterizes the spread of potential energies of the mutated population with respect to temperature, three different statistical regimes have been identified. We suggest that biopolymers found in nature are in a critical regime with g(0) approximately 1-6, corresponding to a broad, but not too broad, phenotypic distribution resembling a truncated Lévy flight. Thus the biopolymer phenotype can be considerably modified in just a few mutations. The proposed model is in good agreement with the experimental distribution of activities determined for a population of single mutants of a group-I ribozyme.

TectoRNA and 'kissing-loop' RNA: atomic force microscopy of self-assembling RNA structures.

RNA molecules have been much less studied by atomic force microscopy (AFM) than have DNA molecules. In this paper, AFM imaging is presented for two different RNA molecules able to self-assemble into complex supramolecular architectures. The first one is a molecular dimer of a 230-nt RNA fragment coming from the RNA genome of a murine leukaemia virus. The monomeric RNA fragment, which appears by AFM as an elongated structure with a mean aspect ratio of 1.4, assembles into a dimer of elongated structures through the formation of a 'kissing-loop' RNA interaction. The second one is a large supramolecular fibre formed of artificial self-assembling RNA molecular units called tectoRNA. The fibre lengths by AFM suggest that there are 50-70 tectoRNA units per fibre. Some methods and limitations are presented for measuring molecular volumes from AFM images.

Unruptured intracranial aneurysms: natural history, clinical outcome, and risks of surgical and endovascular treatment.

BACKGROUND: The management of unruptured intracranial aneurysms is controversial. Investigators from the International Study of Unruptured Intracranial Aneurysms aimed to assess the natural history of unruptured intracranial aneurysms and to measure the risk associated with their repair. METHODS: Centres in the USA, Canada, and Europe enrolled patients for prospective assessment of unruptured aneurysms. Investigators recorded the natural history in patients who did not have surgery, and assessed morbidity and mortality associated with repair of unruptured aneurysms by either open surgery or endovascular procedures. FINDINGS: 4060 patients were assessed-1692 did not have aneurysmal repair, 1917 had open surgery, and 451 had endovascular procedures. 5-year cumulative rupture rates for patients who did not have a history of subarachnoid haemorrhage with aneurysms located in internal carotid artery, anterior communicating or anterior cerebral artery, or middle cerebral artery were 0%, 2. 6%, 14 5%, and 40% for aneurysms less than 7 mm, 7-12 mm, 13-24 mm, and 25 mm or greater, respectively, compared with rates of 2 5%, 14 5%, 18 4%, and 50%, respectively, for the same size categories involving posterior circulation and posterior communicating artery aneurysms. These rates were often equalled or exceeded by the risks associated with surgical or endovascular repair of comparable lesions. Patients' age was a strong predictor of surgical outcome, and the size and location of an aneurysm predict both surgical and endovascular outcomes. INTERPRETATION: Many factors are involved in management of patients with unruptured intracranial aneurysms. Site, size, and group specific risks of the natural history should be compared with site, size, and age-specific risks of repair for each patient.

Dimerization of the 3'UTR of bicoid mRNA involves a two-step mechanism.

The proper localization of bicoid (bcd) mRNA requires cis-acting signals within its 3' untranslated region (UTR) and trans-acting factors such as Staufen. Dimerization of bcd mRNA through intermolecular base-pairing between two complementary loops of domain III of the 3'UTR was proposed to be important for particle formation in the embryo. The participation in the dimerization process of each domain building the 3'UTR was evaluated by thermodynamic and kinetic analysis of various mutated and truncated RNAs. Although sequence complementarity between the two loops of domain III is required for initiating mRNA dimerization, the initial reversible loop-loop complex is converted rapidly into an almost irreversible complex. This conversion involves parts of RNA outside of domain III that promote initial recognition, and dimerization can be inhibited by sense or antisense oligonucleotides only before conversion has proceeded. Injection of the different bcd RNA variants into living Drosophila embryos shows that all elements that inhibit RNA dimerization in vitro prevent formation of localized particles containing Staufen. Particle formation appeared to be dependent on both mRNA dimerization and other element(s) in domains IV and V. Domain III of bcd mRNA could be substituted by heterologous dimerization motifs of different geometry. The resulting dimers were converted into stable forms, independently of the dimerization module used. Moreover, these chimeric RNAs were competent in forming localized particles and recruiting Staufen. The finding that the dimerization domain of bcd mRNA is interchangeable suggests that dimerization by itself, and not the precise geometry of the intermolecular interactions, is essential for the localization process. This suggests that the stabilizing interactions that are formed during the second step of the dimerization process might represent crucial elements for Staufen recognition and localization.

Muc-1, integrin, and osteopontin expression during the implantation cascade in sheep.

The extracellular matrix protein osteopontin (OPN) is a component of histotroph that increases in uterine flushings from pregnant ewes during the peri-implantation period and is localized on the apical surfaces of the uterine luminal epithelium (LE) and conceptus trophectoderm (Tr). The potential involvement of OPN in the implantation adhesion cascade in sheep was investigated by examining temporal, spatial, and potential functional relationships between OPN, Muc-1, and integrin subunits during the estrous cycle and early pregnancy. Immunoreactive Muc-1 was highly expressed at the apical surfaces of uterine luminal (LE) and glandular epithelium (GE) in both cycling and pregnant ewes but was decreased dramatically on LE by Day 9 and was nearly undetectable by Day 17 of pregnancy when intimate contact between LE and Tr begins. In contrast, integrin subunits alpha(v), alpha(4), alpha(5), beta(1), beta(3), and beta(5) were constitutively expressed on conceptus Tr and at the apical surface of uterine LE and GE in both cyclic and early pregnant ewes. The apical expression of these subunits could contribute to the apical assembly of several OPN receptors including the alpha(v)beta(3), alpha(v)beta(1), alpha(v)beta(5), alpha(4)beta(1), and alpha(5)beta(1) heterodimers on endometrial LE and GE, and conceptus Tr in sheep. Functional analysis of potential OPN interactions with conceptus and endometrial integrins was performed on LE and Tr cells in vitro using beads coated with OPN, poly-L-lysine, or recombinant OPN in which the Arg-Gly-Asp sequence was replaced with RGE or RAD. Transmembrane accumulation of talin or alpha-actinin at the apical surface of uterine LE and conceptus Tr cells in contact with OPN-coated beads revealed functional integrin activation and cytoskeletal reorganization in response to OPN binding. These results provide a physiological framework for the role of OPN, a potential mediator of implantation in sheep, as a bridge between integrin heterodimers expressed by Tr and uterine LE responsible for adhesion for initial conceptus attachment.

Ornithine aminotransferase messenger RNA expression and enzymatic activity in fetal porcine intestine.

In most neonatal animals, the small intestinal epithelium is responsible for endogenous arginine production. The ability of neonatal enterocytes to synthesize arginine immediately after birth suggests that the enzymes involved are present prenatally. Pyrroline-5-carboxylate is the common intermediate in the intestinal pathways for the synthesis of citrulline and arginine from both glutamine and proline and is interconverted into ornithine by ornithine aminotransferase (OAT). In this study, OAT enzymatic activity and mRNA expression in the intestine of fetal pigs from 30 to 110 d of gestation were determined. Enzymatic activity (nanomoles per minute per milligram of protein) peaked at d 45 of gestation and increased again between d 60 and 110 of gestation. At 30 and 35 d of gestation, OAT mRNA expression was detected throughout the mucosal epithelium of the small intestine. Throughout the remainder of gestation, OAT expression was notably higher in the villus epithelium than in the crypt epithelium. The presence of OAT in the small intestinal epithelium throughout gestation suggests that the porcine small intestine is capable of interconverting ornithine and pyrroline-5-carboxylate during fetal development. This capability may be important for synthesis of arginine, proline, ornithine, and polyamines for development and metabolic activity of the intestine during gestation or for somatic growth of the fetus.

Keratinocyte growth factor is up-regulated by estrogen in the porcine uterine endometrium and functions in trophectoderm cell proliferation and differentiation.

Keratinocyte growth factor (KGF) is expressed by uterine endometrial epithelial cells during the estrous cycle and during pregnancy in pigs, whereas KGF receptor is expressed in conceptus trophectoderm and endometrial epithelia. In particular, KGF expression in the endometrium is highest on day 12 of pregnancy. This corresponds to the period of maternal recognition of pregnancy in pigs, which is signaled by large amounts of estrogen secreted by conceptus trophectoderm acting on the endometrium. Our hypothesis is that estrogens of conceptus origin stimulate endometrial epithelial KGF expression, and, in turn, secreted KGF stimulates proliferation and differentiation of conceptus trophectoderm. To determine the factors affecting KGF expression in the uterus, endometrial explants from gilts on day 9 of the estrous cycle were cultured in the presence of 17beta-estradiol, catechol estrogens, or progesterone. 17beta-Estradiol stimulated the expression of KGF (P < 0.05), whereas catechol estrogens had no effect (P > 0.05). Between days 9 and 15 of pregnancy, proliferating cell nuclear antigen was abundant in conceptuses, but was barely detectable in uterine endometrial epithelia. To determine the effects of KGF on conceptus trophectoderm, porcine trophectoderm (pTr) cells were treated with recombinant rat KGF (rKGF). rKGF increased the proliferation of pTr cells (P < 0.01) as measured by [(3)H]thymidine incorporation. rKGF elicited phosphorylation of KGF receptor and activated the mitogen-activated protein kinase (ERK1/2) cascade in pTr cells. pTr cell differentiation was affected by rKGF, because it increased expression of urokinase-type plasminogen activator, a marker for differentiation in pTr cells. Collectively, these results indicate that estrogen, the pregnancy recognition signal from the conceptus in pigs, increases uterine epithelial KGF expression, and, in turn, KGF stimulates the proliferation and differentiation of conceptus trophectoderm.

TectoRNA: modular assembly units for the construction of RNA nano-objects.

Structural information on complex biological RNA molecules can be exploited to design tectoRNAs or artificial modular RNA units that can self-assemble through tertiary interactions thereby forming nanoscale RNA objects. The selective interactions of hairpin tetraloops with their receptors can be used to mediate tectoRNA assembly. Here we report on the modulation of the specificity and the strength of tectoRNA assembly (in the nanomolar to micromolar range) by variation of the length of the RNA subunits, the nature of their interacting motifs and the degree of flexibility of linker regions incorporated into the molecules. The association is also dependent on the concentration of magnesium. Monitoring of tectoRNA assembly by lead(II) cleavage protection indicates that some degree of structural flexibility is required for optimal binding. With tectoRNAs one can compare the binding affinities of different tertiary motifs and quantify the strength of individual interactions. Furthermore, in analogy to the synthons used in organic chemistry to synthesize more complex organic compounds, tectoRNAs form the basic assembly units for constructing complex RNA structures on the nanometer scale. Thus, tectoRNA provides a means for constructing molecular scaffoldings that organize functional modules in three-dimensional space for a wide range of applications.

Functional analysis of autocrine and paracrine signalling at the uterine-conceptus interface in pigs.

The complexity of implantation necessitates intimate dialogue between conceptus and maternal cells, and precise coordination of maternal and conceptus signalling events. Maternal and conceptus-derived steroid hormones, growth factors and cytokines, as well as integrins and their ligands, have important and inter-related roles in mediating adhesion between apical aspects of conceptus trophectoderm and maternal uterine luminal epithelium that leads to formation of an epitheliochorial placenta. Integrin receptors appear to play fundamental roles in the implantation cascade and may interact with extracellular matrix molecules and other ligands to transduce cellular signals through autocrine and paracrine mechanisms. Functional in vitro analyses can be used to monitor individual contributions of specific integrin receptors and ligands to the signalling cascades of the maternal-conceptus interface. Integrative studies of implantation in pigs, using in vivo and in vitro approaches, are required to understand conceptus attachment and implantation in this species, and provide valuable opportunities to understand the fundamental mechanisms of implantation in all species.