Selective distortion of the planar group CαC'(O)O to a chiral flat tetrahedron in the amino acid alanine

Based on a Cambridge Structural Database (CSD) search, a meta‐analysis of 116 structures of alanine H₃NC_αH(CH₃)C′(O)O and its derivatives H₃NC_αH(CH₃)C′(O)O(H/R/M), protonated, esterified, or coordinated at the carboxylic group, shows that in the first step of a chirality chain, the L configuration at C_α induces (M) and (P) conformations with respect to rotation around the central C′─C_α bond. In the second step, the (M) and (P) conformations selectively distort the planar carboxylic group C_αC'(O_cis)O_trans to asymmetric flat (R) and (S) tetrahedra. High diastereoselectivities are caused by the two players attraction N…O_cis and repulsion O_trans…C_Me, which work together in (L,M,R) configurations but against each other in (L,P,S) configurations.     

Rotifers,the jelly plankton of freshwater

Jelly plankton (gelata) is a rather fuzzy concept that I attempt to redefine and expand: gelata should have no exo or endoskeleton, have a water content of 92–98%, and should usually be somewhat transparent. Although gelata are usually considered as part of the plankton, there are many cases in which they lead a benthic life, permanently or during part of their life. Gelatinous material can be part of the body, or can be secreted to form a sheath around it, especially in freshwater. Size is not a criterion: gelata individuals or colonies can be from 50 μm to 30 m and more in size. Early development stages (eggs and embryos) shed freely in the environment are not considered true gelata, even if enveloped in jelly. Under these conditions, jelly plankton in freshwater is represented by 20–40 species of cnidarians, by a few rhabdocoelid flatworms, and by up to 200 rotifer species (600+ if the periphytic and benthic species are included, and almost 1,500 species if males are included). Contrary to the ocean, where overfishing will benefit gelata, overfishing in freshwater will favor planktonic crustaceans, and these, in turn competitively suppress rotifers. Guest editors: S. S. S. Sarma, R. D. Gulati, R. L. Wallace, S. Nandini, H. J. Dumont and R. Rico-Martínez Advances in Rotifer Research     

Particulate organic carbon in the estuarine turbidity maxima of the Gironde,Loire and Seine estuaries: origin and lability

A study of the particulate organic carbon (POC) in the estuarine turbidity maxima (ETMs) of the three major French macrotidal estuaries shows that the average contents are 1.5, 3.3 and 3.1% (expressed in % of dry suspended sediment) in the Gironde, Loire and Seine Estuaries, respectively. There is no seasonal variation of POC contents in the Gironde Estuary, whereas, they often increase in the Loire and the Seine Estuaries in spring and summer. The lability of the estuarine particulate organic matter was estimated by two analyses: 1/labile organic matter was measured as the organic carbon loss during incubation tests over one month; 2/ the hydrolysable organic fraction was determined after 6N HCl digestion. The organic fractions of the ETMs are mainly refractory. Any increase in the amount of POC as compared to the background levels (cited above) is always correlated to an increase of organic matter lability. The yearly average fluvial contributions by various particulate organic pools (soil and litter organic matter; organic matter of phytoplanktonic and human origin) that enter the three estuaries were quantified. In the Garonne River, soil and litter are the major (90%) POC sources. In the Loire system, due to the eutrophication of the river water, phytoplankton contributes up to 50% of the total POC load. In the Seine river, soil and litter contribute 70% of the total POC input; POC of human origin is also significant (10%), due to the impact of the City of Paris (10 million inhabitants). The lability of the different types of organic matter ranks as follows: phytoplankton ∼litter > human-origin organic matter >> soil. By combining the POC budgets and the lability of each type of organic fraction, it was possible to explain why the POC of the three ETMs is different and characterizes its refractory vs. labile nature.     

Assessment of temperature effects on beta-aggregation of native and glycated albumin by FTIR spectroscopy and PAGE: relations between structural changes and antioxidant properties

Structural modifications of bovine serum albumin (BSA) induced by heating, and the involvement of glycation of albumin in such processing were studied by using Fourier transform infrared spectroscopy (FTIR) and polyacrylamide gel electrophoresis (PAGE). For native BSA, heating treatments gave rise to beta structures which were amplified to the detriment of alpha-helix form, and which were associated with increased aggregation. A very high correlation was obtained between FTIR Amide I band evolution and aggregation rate parameters, showing the contribution of beta-form in aggregates formation. We further assessed the effect of glycation on protein sensibility to heating treatments. A reduction of conformational changes and aggregation processes was demonstrated for the glycated form of the protein. The antioxidant properties of albumin were evaluated using two different techniques assessing metal binding and free radical neutralizing capacities of the protein. Associations between structural changes in BSA induced by the thermal treatment and its antioxidant activities were established.     

Spatial variability in digenean metacercariae infection of 0-group common sole Solea solea among nurseries along the French Atlantic coast

We sampled 0-group sole juveniles (N = 174) in September and October 2003 in 9 major nurseries located along the French Atlantic coast (English Channel and Bay of Biscay). 0-group sole were infected with 3 genera of digenean metacercariae, Timoniella spp. (Acanthostomidae), Prosorhynchus crucibulum and Prosorhynchus sp. A (Bucephalidae), Podocotyle sp. (Opecoelidae), and an unidentified species of Digenea. Parasite infection levels in the English Channel nurseries were lower, and the community composition was different from Bay of Biscay nurseries. We hypothesize that the difference between geographic areas was due to differences in first intermediate host communities and, to a lesser extent, because of lower temperatures in the English Channel compared to the Bay of Biscay. For the Bay of Biscay nurseries, mean total parasite abundance was strongly negatively correlated with mean annual river flow. This relationship may be the result of the more upstream location of 0-group sole nurseries in estuaries so that fish were further from local points of parasite transmission than those in embayed nurseries. Digenean metacercariae load may be influenced by 3 major local factors, i.e. abundance and proximity of the first intermediate hosts and cercariae dispersal capacities.     

The Saanen goat as an animal model for post-laryngectomy research: practical implications

A modern way of voice rehabilitation after total laryngectomy includes the use of shunt valves and tracheostoma valves. Problems of fixation to the surrounding tissue are a major drawback in the use of the shunt valve, heat and moisture exchange (HME) filters and, especially, the tracheostoma valve. To solve these problems different tissue connectors were developed. The main objective was to test the feasibility of these prototypes in a new animal model. Here we discuss the results, problems and complications of the selected Saanen goat model. In this prospective laboratory study, 19 healthy adult female Saanen goats (Capra hircus) were used and observed post-surgically for 12 weeks. Selection criteria such as comparable anatomy to humans and easy handling were used for animal model development. Also a literature search using the Medline and the ISI Web of Science databases was performed. The anatomy of the Saanen goat was investigated in a separate postmortem study. Surgery consisted of a laryngotracheal separation and implantation of a tracheo-oesophageal and tracheostoma tissue connector with fibrin tissue glue. Postoperative care consisted of frequent stoma care, monitoring appetite, weight, vital signs and administration of antibiotics, analgesics and mucolytic agents. All animals survived the surgical procedure. However, postoperative care was extensive, labour intensive and was accompanied by several complications. Eleven animals died spontaneously before the end of the experiment. The tracheostoma tissue connector caused signs of local infection in all cases. There was no evidence of infection around the tracheo-oesophageal tissue connector in 18 cases. It was concluded that the use of goats in this tracheostoma model was associated with major complications and should, therefore, only be used for short-term experiments with intensive care. Additional research is needed to see if clinical application of the tissue connectors is possible in the future.     

A genomic map of climate adaptation in Mediterranean cattle breeds

Domestic species such as cattle (Bos taurus taurus and B. t. indicus) represent attractive biological models to characterize the genetic basis of short‐term evolutionary response to climate pressure induced by their post‐domestication history. Here, using newly generated dense SNP genotyping data, we assessed the structuring of genetic diversity of 21 autochtonous cattle breeds from the whole Mediterranean basin and performed genome‐wide association analyses with covariables discriminating the different Mediterranean climate subtypes. This provided insights into both the demographic and adaptive histories of Mediterranean cattle. In particular, a detailed functional annotation of genes surrounding variants associated with climate variations highlighted several biological functions involved in Mediterranean climate adaptation such as thermotolerance, UV protection, pathogen resistance or metabolism with strong candidate genes identified (e.g., NDUFB3, FBN1, METTL3, LEF1, ANTXR2 and TCF7). Accordingly, our results suggest that main selective pressures affecting cattle in Mediterranean area may have been related to variation in heat and UV exposure, in food resources availability and in exposure to pathogens, such as anthrax bacteria (Bacillus anthracis). Furthermore, the observed contribution of the three main bovine ancestries (indicine, European and African taurine) in these different populations suggested that adaptation to local climate conditions may have either relied on standing genomic variation of taurine origin, or adaptive introgression from indicine origin, depending on the local breed origins. Taken together, our results highlight the genetic uniqueness of local Mediterranean cattle breeds and strongly support conservation of these populations.     

Sucrose prevents protein fibrillation through compaction of the tertiary structure but hardly affects the secondary structure

Amyloid fibers, implicated in a wide range of diseases, are formed when proteins misfold and stick together in long rope‐like structures. As a natural mechanism, osmolytes can be used to modulate protein aggregation pathways with no interference with other cellular functions. The osmolyte sucrose delays fibrillation of the ribosomal protein S6 leading to softer and less shaped‐defined fibrils. The molecular mechanism used by sucrose to delay S6 fibrillation was studied based on the two‐state unfolding kinetics of the secondary and tertiary structures. It was concluded that the delay in S6 fibrillation results from stabilization and compaction of the slightly expanded tertiary native structure formed under fibrillation conditions. Interestingly, this compaction extends to almost all S6 tertiary structure but hardly affects its secondary structure. The part of the S6 tertiary structure that suffered more compaction by sucrose is known to be the first part to unfold, indicating that the native S6 has entered the unfolding pathway under fibrillation conditions. Proteins 2015; 83:2039–2051. © 2015 Wiley Periodicals, Inc.     

Recombinant Factor VIIa Reduces Bleeding after Blunt Liver Injury in a Pig Model of Dilutional Coagulopathy under Severe Hypothermia

Background

Recombinant factor VIIa (rFVIIa) is registered for use in haemophilia with inhibitors and other rare bleeding disorders, but has also been used in various other clinical conditions to terminate life-threatening bleeding. Underlying conditions (e.g. coagulopathy) and dosing may affect treatment efficacy. The objective of the present study was to evaluate the impact of increasing doses of rFVIIa on blood loss and coagulation assays in haemodiluted and hypothermic pigs undergoing blunt liver injury.

Methods

A grade III blunt liver injury was induced in 28 pigs after 70% haemodilution and cooling to 32.6–33.4°C. Ten minutes after trauma, animals randomly received placebo or 90, 180 or 360 μg/kg rFVIIa. Global coagulation parameters, thromboelastometry (TEM) and plasma thrombin generation (TG) were determined at different time points during the observation period of 120 minutes.

Results

Total blood loss was significantly lower following 90 μg/kg rFVIIa (1206 [1138–1470] mL) relative to placebo (2677 [2337–3068] mL; p<0.05), with no increased effect with higher dose levels of rFVIIa. Following trauma and haemodilution, coagulation was impaired relative to baseline in both TEM and TG analysis. At 60 and 120 minutes after trauma, TEM variables improved in the rFVIIa-treated animals compared with the placebo group. Similarly, rFVIIa improved coagulation kinetics in TG. As was observed with blood loss, no significant effect between different rFVIIa dose levels was found in TEM or TG. Macro- and microscopic post-mortem examination did not reveal any signs of thromboembolic events.

Conclusion

Early administration of 90 μg/kg rFVIIa reduced blood loss in pigs undergoing blunt liver injury even after severe haemodilution and hypothermia, with no further effect of higher dose levels. Coagulation assays showed impaired coagulation in coagulopathic animals, with a dose-independent improvement in animals treated with rFVIIa.     

Cervical Lymph Nodes as a Selective Niche for Brucella during Oral Infections

Cervical lymph nodes (CLN) are the first lymph nodes encountered by material taking the oral route. To study their role in orally acquired infections, we analyzed 307 patients of up to 14 years treated in the university clinic of Skopje, Macedonia, for brucellosis, a zoonotic bacterial disease frequently acquired by ingestion of contaminated dairy products. From these children, 36% had lymphadenopathy. Among orally infected children, lymphadenopathy with CLN being the only lymph nodes affected was significantly more frequent as compared to those infected by contact with animals (83% vs. 63%), suggesting a possible involvement of CLN during orally acquired human brucellosis. Using a murine model where bacteria are delivered into the oral cavity, we show that Brucella quickly and selectively colonize the CLN where they proliferate and persist over long periods of time for up to 50 days post-infection. A similar efficient though less specific drainage to CLN was found for Brucella, Salmonella typhimurium and fluorescent microspheres delivered by gavage, a pathway likely representing a mixed infection mode of intragastric and oral infection, suggesting a central pathway of drained material. Microspheres as well as bacteria drained to CLN predominately reside in cells expressing CD68 and no or low levels of CD11c. Even though no systemic response could be detected, Brucella induced a locally restricted inflammatory reaction with increased expression levels of interferon γ, interleukin (IL)-6, IL-12, granzyme B and a delayed induction of Nos2. Inflammation led to pronounced lymphadenopathy, infiltration of macrophages/monocytes expressing high levels of major histocompatibility complex II and to formation of epitheloid granulomas. Together, these results highlight the role of CLN in oral infections as both, an initial and efficient trap for bacterial invaders and as possible reservoir for chronic pathogens. They likewise cast a new light on the significance of oral routes for means of vaccination.