Aspergillus tubingensis Endocarditis: A Case Report and Review of the Literature

Mycopathologia - Aspergillus endocarditis is a rare infection that may affect immunocompetent patients following heart valve replacement or heart surgery. We report the case of a 39 year...     

The intrinsic contributions of tyrosine, serine, glycine and arginine to the affinity and specificity of antibodies

Synthetic antibody libraries with restricted chemical diversity were used to explore the intrinsic contributions of four amino acids (Tyr, Ser, Gly and Arg) to the affinity and specificity of antigen recognition. There was no correlation between nonspecific binding and the content of Tyr, Ser or Gly in the antigen-binding site, and in fact, the most specific antibodies were those with the highest Tyr content. In contrast, Arg content was clearly correlated with increased nonspecific binding. We combined Tyr, Ser and Gly to generate highly specific synthetic antibodies with affinities in the subnanomolar range, showing that the high abundance of Tyr, Ser and Gly in natural antibody germ line sequences reflects the intrinsic capacity of these residues to work together to mediate antigen recognition. Despite being a major functional contributor to co-evolved protein-protein interfaces, we find that Arg does not contribute generally to the affinity of naïve antigen-binding sites and is detrimental to specificity. Again, this is consistent with studies of natural antibodies, which have shown that nonspecific, self-reactive antibodies are rich in Arg and other positively charged residues. Our findings suggest that the principles governing naïve molecular recognition differ from those governing co-evolved interactions. Analogous studies can be designed to explore the roles of the other amino acids in molecular recognition. Results of such studies should illuminate the basic principles underlying natural protein-protein interactions and should aid the design of synthetic binding proteins with functions beyond the scope of natural proteins.     

Mise en évidence de structures internes des chromosomes par une technique nouvelle (Scanning video)

The basic principle of the technique is the analysis of photomicrographs (light microscope) with a system of closed television chain. By suitable settings of the video scanning (e.g. black and white level, contrast) it is possible to bring out some particular structures of chromosomes, by a kind of optical density selection. — Chromosomes in mitosis and meiosis from different species (man, rat, sloth) have been studied up to now; all of them show a similar inner organization. The double spiralization of each chromatid appears clearly. Each chromonema contains a more or less dense heap of fine loops; these appear to be made of a folded fibril of 200–300 Å thickness. In less condensed zones of the chromonema, e.g. in uncoiled parts, four filaments arranged in two more or less twisted pairs are clearly distinguishable; these filaments seem to correspond to 1/8 chromatid. From our first investigations it seems that the inner fibrils of the chromosomes are organized according to one of the DuPraw's models (combined transverse and longitudinal folding with quaternary coiling). — Some arguments are proposed and discussed as to explain how it is possible to reach such a high resolution with a conventional light microscope.

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Recherches effectuées avec l'aide du Fonds de la Recherche Scientifique Médicale (Belgique).     

The Controversy on Fish Pain: A Veterinarian’s Perspective

Fish welfare is still a relatively new field. As such, regulations and protocols to ensure fish welfare are currently limited and vary considerably in different jurisdictions. This is in part because of the ongoing controversy as to whether or not fish feel pain. This controversy has persisted for several years, yet veterinarians have been mostly absent from the discussion so far. This essay aims to address this issue. Here, it is argued that while this controversy has its place, it is unlikely to be resolved in the near future. Fish welfare could instead be improved by pursuing more clinically applicable research to increase knowledge of fishes’ behavior and physiology. Such research would assist in learning the optimal environment for their specific needs, as well as compiling some verified indicators of pain in fish. This would then lead to improved studies that could help to determine if and when analgesic drugs can be beneficial in fish, as they are in many other species.     

Combination of active and inactive siRNA targeting the mitotic kinesin Eg5 impairs silencing efficiency in several cancer cell lines

Gene silencing by RNA interference (RNAi) has proven to be a powerful tool for investigating gene function in mammalian cells. Combination of several short interfering RNA (siRNA) targeting the same gene is commonly used to improve RNA interference. However, in contrary to the well-described mechanism of RNAi, efficiency of single siRNA compared to pool remains poorly documented. We addressed this issue using several active and inactive siRNA targeting Eg5, a kinesin-related motor involved in mitotic spindle assembly. These siRNA, used alone or in combination, were tested for their silencing efficiency in several cancer cell lines. Here we show that presence of inactive Eg5 siRNA in a pool dramatically decreases knockdown efficacy in a cell line- and dose-dependent manner. Lack of inhibition by unrelated siRNA suggests that a competition may occur during siRNA incorporation into RNA-induced silencing complexes (RISCs) along with the target mRNA. Altogether, our results, which need to be confirmed with additional inactive siRNA, indicate that combination of siRNA may not increase but instead decrease silencing efficiency.     

Optimal pedalling velocity characteristics during maximal and submaximal cycling in humans

The aim of this study was to compare optimal pedalling velocities during maximal (OVM) and submaximal (OVSM) cycling in human, subjects with different training backgrounds. A group of 22 subjects [6 explosive (EX), 6 endurance (EN) and 10 non-specialised subjects] sprint cycled on a friction-loaded ergometer four maximal sprints lasting 6 s each followed by five 3-min periods of steady-state cycling at 150 W with pedalling frequencies varying from 40 to 120 rpm. The OVM and OVSM were defined as the velocities corresponding to the maximal power production and the lowest oxygen consumption, respectively. A significant linear relationship (r2 = 0.52, P < 0.001) was found between individual OVM [mean 123.1 (SD 11.2) rpm] and OVSM [mean 57.0 (SD 4.9) rpm, P < 0.001] values, suggesting that the same functional properties of leg extensor muscles influence both OVM and OVSM. Since EX was greater than EN in both OVM and OVSM (134.3 compared to 110.9 rpm and 60.8 compared to 54.0 rpm, P < 0.01 and P < 0.05, respectively) it could be hypothesised that the distribution of muscle fibre type plays an important role in optimising both maximal and submaximal cycling performance.     

New simian immunodeficiency virus infecting De Brazza's monkeys (Cercopithecus neglectus): evidence for a cercopithecus monkey virus clade

Nearly complete sequences of simian immunodeficiency viruses (SIVs) infecting 18 different nonhuman primate species in sub-Saharan Africa have now been reported; yet, our understanding of the origins, evolutionary history, and geographic distribution of these viruses still remains fragmentary. Here, we report the molecular characterization of a lentivirus (SIVdeb) naturally infecting De Brazza's monkeys (Cercopithecus neglectus). Complete SIVdeb genomes (9,158 and 9227 bp in length) were amplified from uncultured blood mononuclear cell DNA of two wild-caught De Brazza's monkeys from Cameroon. In addition, partial pol sequences (650 bp) were amplified from four offspring of De Brazza's monkeys originally caught in the wild in Uganda. Full-length (9068 bp) and partial pol (650 bp) SIVsyk sequences were also amplified from Sykes's monkeys (Cercopithecus albogularis) from Kenya. Analysis of these sequences identified a new SIV clade (SIVdeb), which differed from previously characterized SIVs at 40 to 50% of sites in Pol protein sequences. The viruses most closely related to SIVdeb were SIVsyk and members of the SIVgsn/SIVmus/SIVmon group of viruses infecting greater spot-nosed monkeys (Cercopithecus nictitans), mustached monkeys (Cercopithecus cephus), and mona monkeys (Cercopithecus mona), respectively. In phylogenetic trees of concatenated protein sequences, SIVdeb, SIVsyk, and SIVgsn/SIVmus/SIVmon clustered together, and this relationship was highly significant in all major coding regions. Members of this virus group also shared the same number of cysteine residues in their extracellular envelope glycoprotein and a high-affinity AIP1 binding site (YPD/SL) in their p6 Gag protein, as well as a unique transactivation response element in their viral long terminal repeat; however, SIVdeb and SIVsyk, unlike SIVgsn, SIVmon, and SIVmus, did not encode a vpu gene. These data indicate that De Brazza's monkeys are naturally infected with SIVdeb, that this infection is prevalent in different areas of the species' habitat, and that geographically diverse SIVdeb strains cluster in a single virus group. The consistent clustering of SIVdeb with SIVsyk and the SIVmon/SIVmus/SIVgsn group also suggests that these viruses have evolved from a common ancestor that likely infected a Cercopithecus host in the distant past. The vpu gene appears to have been acquired by a subset of these Cercopithecus viruses after the divergence of SIVdeb and SIVsyk.     

The embryonic leaf identity gene FUSCA3 regulates vegetative phase transitions by negatively modulating ethylene-regulated gene expression in Arabidopsis

Background

Protein kinase CK2 is a pleiotropic serine/threonine protein kinase with hundreds of reported substrates, and plays an important role in a number of cellular processes. The cellular functions of Plasmodium falciparum CK2 (PfCK2) are unknown. The parasite's genome encodes one catalytic subunit, PfCK2??, which we have previously shown to be essential for completion of the asexual erythrocytic cycle, and two putative regulatory subunits, PfCK2??1 and PfCK2??2.

Results

We now show that the genes encoding both regulatory PfCK2 subunits (PfCK2??1 and PfCK2??2) cannot be disrupted. Using immunofluorescence and electron microscopy, we examined the intra-erythrocytic stages of transgenic parasite lines expressing hemagglutinin (HA)-tagged catalytic and regulatory subunits (HA-CK2??, HA-PfCK2??1 or HA-PfCK2??2), and localized all three subunits to both cytoplasmic and nuclear compartments of the parasite. The same transgenic parasite lines were used to purify PfCK2??1- and PfCK2??2-containing complexes, which were analyzed by mass spectrometry. The recovered proteins were unevenly distributed between various pathways, with a large proportion of components of the chromatin assembly pathway being present in both PfCK2??1 and PfCK2??2 precipitates, implicating PfCK2 in chromatin dynamics. We also found that chromatin-related substrates such as nucleosome assembly proteins (Naps), histones, and two members of the Alba family are phosphorylated by PfCK2?? in vitro.

Conclusions

Our reverse-genetics data show that each of the two regulatory PfCK2 subunits is required for completion of the asexual erythrocytic cycle. Our interactome study points to an implication of PfCK2 in many cellular pathways, with chromatin dynamics being identified as a major process regulated by PfCK2. This study paves the way for a kinome-wide interactomics-based approach to elucidate protein kinase function in malaria parasites.