Ruminal fermentation characteristics and microbial nitrogen assimilation in sheep fed differently composed grass silages

The investigation aimed at examining if the composition of grassland silage affects the microbial nitrogen assimilation in the rumen of sheep. The silages were made of vegetative summer re-growths consisting of 48% grasses, 28% legumes and 24% other forbs (GCF) or of pure grass (G). Silage GCF contained more intermediately degradable non-structural and less slowly degradable carbohydrates, more crude protein (CP), a narrower ratio between slow and very slow degradable nitrogen (N), and exhibited higher in situ degradability of organic matter and CP than Silage G. Four adult wethers equipped with rumen fistulae were used in a two factorial trial. Feed was offered either as silage alone or as a mixture of silage and barley (60:40). Microbial N was estimated using continuous intraruminal 15N infusion and measurement of 15N-enrichment in microbes isolated from rumen liquor samples. With the exception of trends for ruminal butyrate concentrations, no interactions were detected between silage and barley feeding. Sheep receiving Silage GCF exhibited larger diurnal fluctuations of ammonia, and produced more microbial N (p < 0.05) than sheep on Silage G. Feeding the silages with barley decreased ruminal pH and elevated the concentrations of butyrate (p < 0.05). The 15N incorporation into microbial N was reduced by barley feeding (p < 0.05) along with a trend to accelerated rumen fluid turnover, resulting in similar microbial N yields as found in sheep receiving silage without barley. It is concluded that the larger and better balanced amounts of intermediately degradable carbohydrate- and N-containing fractions favoured the ruminal microbial protein synthesis in sheep consuming Silage GCF instead of Silage G.     

Blood volume-monitored regulation of ultrafiltration in fluid-overloaded hemodialysis patients: study protocol for a randomized controlled trial

Background

Governments and donors all over Africa are searching for sustainable, affordable and cost-effective ways to improve the quality of malaria case management. Widespread deficiencies have been reported in the prescribing and counselling practices of health care providers treating febrile patients in both public and private health facilities. Cameroon is no exception with low levels of adherence to national guidelines, the frequent selection of non-recommended antimalarials and the use of incorrect dosages. This study evaluates the effectiveness and cost-effectiveness of introducing two different provider training packages, alongside rapid diagnostic tests (RDTs), designed to equip providers with the knowledge and practical skills needed to effectively diagnose and treat febrile patients. The overall aim is to target antimalarial treatment better and to facilitate optimal use of malaria treatment guidelines.

Methods/Design

A 3-arm stratified, cluster randomized trial will be conducted to assess whether introducing RDTs with provider training (basic or enhanced) is more cost-effective than current practice without RDTs, and whether there is a difference in the cost effectiveness of the provider training interventions. The primary outcome is the proportion of patients attending facilities that report a fever or suspected malaria and receive treatment according to malaria guidelines. This will be measured by surveying patients (or caregivers) as they exit public and mission health facilities. Cost-effectiveness will be presented in terms of the primary outcome and a range of secondary outcomes, including changes in provider knowledge. Costs will be estimated from a societal and provider perspective using standard economic evaluation methodologies.

Trial Registration

ClinicalTrials.gov: NCT00981877     

Competitive cellular uptake of nanoparticles made from polystyrene, poly(methyl methacrylate), and polylactide

The uptake behavior of negatively charged fluorescent nanoparticles made from different polymers (PS, PMMA, and PLLA) is studied on HeLa cells. All particles are obtained by the miniemulsion process using sodium dodecylsulfate as anionic surfactant. The size of the particles is in the range 105-125 nm. Cell uptake is analyzed by flow cytometry and reveals a higher uptake of PLLA particles compared to PMMA and PS particles. In competitive uptake studies two different types of particles are co-incubated with the HeLa cells; the results indicate a mutual influence of the particles on their uptake behavior. A reduced internalization of PLLA particles in the presence of PS particles is observed, although neither the co-incubation of PMMA and PLLA nor of PMMA and PS shows similar effect.     

Investigation of protein-RNA interactions by mass spectrometry--Techniques and applications

Protein-RNA complexes play many important roles in diverse cellular functions. They are involved in a wide variety of different processes in growth and differentiation at the various stages of the cell cycle. As their function and catalytic activity are directly coupled to the structural arrangement of their components--proteins and ribonucleic acids--the investigation of protein-RNA interactions is of great functional and structural importance. Here we discuss the most prominent examples of protein-RNA complexes and describe some frequently used purification strategies. We present various techniques and applications of mass spectrometry to study protein-RNA complexes. We discuss the analysis of intact complexes as well as proteomics-based and crosslinking-based approaches in which proteins are cleaved into smaller peptides. This article is part of a Special Section entitled: Understanding genome regulation and genetic diversity by mass spectrometry.     

BH3-only protein expression determines hepatocellular carcinoma response to sorafenib-based treatment

Hepatocellular carcinoma (HCC) represents a global health challenge with limited therapeutic options. Anti-angiogenic immune checkpoint inhibitor-based combination therapy has been introduced for progressed HCC, but improves survival only in a subset of HCC patients. Tyrosine-kinase inhibitors (TKI) such as sorafenib represent an alternative treatment option but have only modest efficacy. Using different HCC cell lines and HCC tissues from various patients reflecting HCC heterogeneity, we investigated whether the sorafenib response could be enhanced by combination with pro-apoptotic agents, such as TNF-related apoptosis-inducing ligand (TRAIL) or the BH3-mimetic ABT-737, which target the death receptor and mitochondrial pathway of apoptosis, respectively. We found that both agents could enhance sorafenib-induced cell death which was, however, dependent on specific BH3-only proteins. TRAIL augmented sorafenib-induced cell death only in NOXA-expressing HCC cells, whereas ABT-737 enhanced the sorafenib response also in NOXA-deficient cells. ABT-737, however, failed to augment sorafenib cytotoxicity in the absence of BIM, even when NOXA was strongly expressed. In the presence of NOXA, BIM-deficient HCC cells could be in turn strongly sensitized for cell death induction by the combination of sorafenib with TRAIL. Accordingly, HCC tissues sensitive to apoptosis induction by sorafenib and TRAIL revealed enhanced NOXA expression compared to HCC tissues resistant to this treatment combination. Thus, our results suggest that BH3-only protein expression determines the treatment response of HCC to different sorafenib-based drug combinations. Individual profiling of BH3-only protein expression might therefore assist patient stratification to certain TKI-based HCC therapies.Subject terms: Cancer, Diseases     

Sustained delayed-type hypersensitivity reaction after in vivo priming but successful induction of unresponsiveness after adoptive transfer of CD4+ effector T cells

Potential reasons for weak effects of oral tolerance in the primed immune system are still under discussion. In the present study, impacts of oral antigen uptake were studied in adoptive transfer models using T cell receptor transgenic CD4(+) T cells allowing analysis of antigen-specific donor cells on single cell level. After in vivo priming and subsequent feeding, an antigen-specific delayed-type hypersensitivity reaction was sustained. Concomitantly, donor cells preferentially found in the draining lymph nodes remained at equal numbers. In contrast, adoptively transferred Th1 cells that migrated preferentially into spleen and liver became fewer upon feeding accompanied by a suppressed delayed-type hypersensitivity reaction. Thus, antigen-experienced cells did not seem to become generally resistant to tolerogenic stimuli. Our data suggest that besides a permanent inflammatory stimulus provided by the persisting antigen, diverse tissue distribution of in vivo-induced compared to adoptively transferred effector cells might interfere with oral tolerance in the experienced immune system.     

Dopaminergic system dysregulation in the mrsk2_KO mouse, an animal model of the Coffin-Lowry syndrome

The Coffin-Lowry syndrome, a rare syndromic form of X-linked mental retardation, is caused by loss-of-function mutations in the hRSK2 (RPS6KA3) gene. To further investigate RSK2 (90-kDa ribosomal S6 kinase) implication in cognitive processes, a mrsk2_KO mouse has previously been generated as an animal model of Coffin-Lowry syndrome. The aim of the present study was to identify possible neurochemical dysregulation associated with the behavioral and morphological abnormalities exhibited by mrsk2_KO mice. A cortical dopamine level increase was found in mrsk2_KO mice that was accompanied by an over-expression of dopamine receptor of type 2 and the dopamine transporter. We also detected an increase of total and phosphorylated extracellular regulated kinase that may be responsible for the increased level of tyrosine hydroxylase phosphorylation also observed. By taking into consideration previously reported data, our results strongly suggest that the dopaminergic dysregulation in mrsk2_KO mice may be caused, at least in part, by tyrosine hydroxylase hyperactivity. This cortical hyperdopaminergia may explain some non-cognitive but also cognitive alterations exhibited by mrsk2_KO mice.     

Macrophages driven to a novel state of activation have anti-inflammatory properties in mice

Recurrent episodes of inflammation underlie numerous pathologies, notably those of inflammatory bowel diseases. In this study, we describe a population of macrophages in a novel state of activation that mitigates colitis in mice. The cells responsible for this effect, called IFN-gamma-stimulated monocyte-derived cells (IFNgamma-MdC), derive from mouse spleen, blood, and bone marrow monocytes and are distinguished from known macrophage populations by mode of generation, cell surface phenotype, and function. IFNgamma-MdC only arise when macrophages are cultivated in the presence of CD40L-expressing CD4+ T cells, M-CSF, and IFN-gamma. IFNgamma-MdC express markers including F4/80, CD11b/c, CD86, and CD274; they are negative for CD4, CD8, Gr1, CD19, CD80, and CD207. Functionally, IFNgamma-MdC are defined by their capacity to enrich cocultured T cell populations for CD4+CD25+Foxp3+ regulatory cells; this enrichment, constituting up to 60% or more of residual lymphocytes, is attributed to an expansion, but also to a cell contact and caspase-dependent depletion of activated T cells. In mice, IFNgamma-MdC delivered i.v. traffic to gut-associated peripheral lymphoid tissues, including the mesenteric lymph nodes, Peyer's patches, and colonic mucosa, and promote the clinical and histological resolution of chronic colitis. We conclude that IFNgamma-MdC represent macrophages in a novel state of activation, possessing multiple T cell-suppressive effects with therapeutic potential for the treatment of autoimmune inflammation.     

Cold hardiness of Apteropanorpa tasmanica Carpenter (Mecoptera: Apteropanorpidae)

There are very few investigations of cold hardiness in native Australian insects, and no such studies on insects from Tasmania. The Apteropanorpidae is a family of wingless Mecoptera endemic to Tasmania, comprising four described species that can be active in winter. In this study, we used infrared video thermography to investigate the physiological and behavioural responses of Apteropanorpa tasmanica to fast (0.3 degrees Cmin(-1)) and slow (0.03 degrees Cmin(-1)) rates of temperature reduction down to -10 degrees C. No adults survived cooling to -10 degrees C at either cooling rate. Mean supercooling points (SCPs) from fast cooling were -7.0 and -4.6 degrees C in 2002 and 2003, respectively. Ice nucleation always began in the abdomen, however, the position of nucleation within the abdomen varied between individuals. There was no relationship between SCP and body length, and no significant difference in SCPs between males and females. Stress-induced fast walking began when insects reached approximately -1.5 degrees C. Cooling rate did not affect the SCP or the temperature at which the behavioural stress response began. Adults survived for only short periods of time in the supercooled state; however they survived in the laboratory for up to 60 days at 4 degrees C, indicating their longevity at more favourable temperatures. Members of the Apteropanorpidae are adapted to the relatively warm, maritime climate currently influencing Tasmania.