Molecular models of NS3 protease variants of the Hepatitis C virus

Background  

Hepatitis C virus (HCV) currently infects approximately three percent of the world population. In view of the lack of vaccines against HCV, there is an urgent need for an efficient treatment of the disease by an effective antiviral drug. Rational drug design has not been the primary way for discovering major therapeutics. Nevertheless, there are reports of success in the development of inhibitor using a structure-based approach. One of the possible targets for drug development against HCV is the NS3 protease variants. Based on the three-dimensional structure of these variants we expect to identify new NS3 protease inhibitors. In order to speed up the modeling process all NS3 protease variant models were generated in a Beowulf cluster. The potential of the structural bioinformatics for development of new antiviral drugs is discussed.     

Study of response of Swiss Webster mice to light subunit of mushroom tyrosinase

The light subunit of mushroom, Agaricus bisporus, tyrosinase (LSMT), has been identified as an extrinsic component of the enzyme. Its function is unknown, but it can cross an epithelial cell layer, which suggests that it can be absorbed by the intestine. A similar capability has been demonstrated for the HA-33 component of the progenitor toxin from Clostridium botulinum, which is the closest structural homolog of LSMT. Unlike HA-33, LSMT appears to be non-immunogenic as shown by preliminary tests in Swiss Webster mice. We investigated the immunogenicity and histopathology of LSMT in mice to determine its safety in vivo. LSMT did not evoke generation of antibodies after prolonged periods of intraperitoneal administration. Histopathological observations confirmed the absence of responses in organs after twelve weekly administrations of LSMT. We found that LSMT is not toxic and is less immunogenic than the C. botulinum HA-33 protein, which supports further research and development for pharmaceutical application.     

A Dutch guideline for the treatment of scoliosis in neuromuscular disorders

Background

Children with neuromuscular disorders with a progressive muscle weakness such as Duchenne Muscular Dystrophy and Spinal Muscular Atrophy frequently develop a progressive scoliosis. A severe scoliosis compromises respiratory function and makes sitting more difficult. Spinal surgery is considered the primary treatment option for correcting severe scoliosis in neuromuscular disorders. Surgery in this population requires a multidisciplinary approach, careful planning, dedicated surgical procedures, and specialized after care.

Methods

The guideline is based on scientific evidence and expert opinions. A multidisciplinary working group representing experts from all relevant specialties performed the research. A literature search was conducted to collect scientific evidence in answer to specific questions posed by the working group. Literature was classified according to the level of evidence.

Results

For most aspects of the treatment scientific evidence is scarce and only low level cohort studies were found. Nevertheless, a high degree of consensus was reached about the management of patients with scoliosis in neuromuscular disorders. This was translated into a set of recommendations, which are now officially accepted as a general guideline in the Netherlands.

Conclusion

In order to optimize the treatment for scoliosis in neuromuscular disorders a Dutch guideline has been composed. This evidence-based, multidisciplinary guideline addresses conservative treatment, the preoperative, perioperative, and postoperative care of scoliosis in neuromuscular disorders.     

A note on pre-spawning maternal cortisol exposure in farmed Atlantic salmon and its impact on the behaviour of offspring in response to a novel environment

This study was designed to assess whether pre-spawning maternal cortisol administration and an additional stressor, here modelled by mild hyperthermia experienced during incubation, affected the behavioural responses of farmed Atlantic salmon (Salmo salar) offspring when exposed to a novel environment. Mature females were injected prior to stripping with 0, 50 or 100 mg/kg cortisol. After fertilization the eggs were kept at incubation temperatures of either 8 or 10 °C. Four months after hatching, the fish were introduced to a novel environment, i.e. an aquarium (38 l). In each trial two individuals originating from different exposure groups were video recorded and their activity (number of turns, time spent not swimming, time spent at the bottom) and feeding behaviour were measured. Maternal cortisol increased time spent non-swimming and also time spent at the bottom, while cortisol and temperature together additionally affected number of turns. To the best of our knowledge this study is the first in fish to demonstrate a possible link between the maternal endocrinological state at spawning and the offspring's behavioural reactivity to novel situations.     

Design and performance of a rugged standard operative temperature thermometer for avian studies

The lack of a truly satisfactory sensor which can characterize the thermal environment at the spatial scale experienced by small endotherms has hindered study of their thermoregulatory behavior. We describe a general design for a rugged, easily constructed sensor to measure standard operative temperature, T_es. We present specific designs for adult dark-eyed juncos (Junco hyemalis) and hatchling mallards (Anas platyrhynchos). Sensor response was stable and repeatable (±1.4%) over the course of several months. Over the range of conditions for which validation data were available (variable air temperature and wind with negligible net radiation), sensors predicted the mean net heat production of live animals to within ±0.023 W (equivalent to ±1°C at T_es=15°C). The main limit on accuracy was scatter in the data on metabolism and evaporative water loss in live animals. These sensors are far more rugged and easily constructed than the heated taxidermic mounts previously used to measure T_es. These characteristics facilitate the use of significant numbers of sensors in thermal mapping studies of endotherms.     

Characterisation and subcellular localisation of the GroEL-like and DnaK-like proteins isolated from Fusobacterium nucleatum ATCC 10953

Fusobacterium nucleatum is associated with periodontitis in humans, and is a central member of the dental biofilm. Heat shock proteins (HSPs) of many different bacteria have been considered to play important roles during inflammations and infections. We have identified and characterised the HSP60 and HSP70, the Escherichia coli GroEL and DnaK homologues, respectively, in F. nucleatum ATCC 10953. The N-terminal 22 amino acid residues of HSP60 exhibited up to 63.6% identity with members of the HSP60 heat shock protein family of some selected bacterial species, while the N-terminal of 25 residues of HSP70 revealed up to 80% identity with members of the HSP70 family. The subcellular localisation of HSP60 and HSP70 was analysed by immunoblotting of bacterial cell fractions and immunoelectron microscopy of whole cells. HSP60 and HSP70 were localised in the cytosol, associated with membranes and extracellular fractions. These results are consistent with localisation for HSPs found in other micro-organisms, which further lead to the suggestion of a potential role in the pathogenesis of infectious diseases.     

Statistical and procedural issues in the use of heated taxidermic mounts

Studies using mounts have an inherently nested error structure; calibration and standardization should use the appropriate procedures and statistics. One example is that individual mount differences are nested within morphological factors related to species, age, or gender; without replication, mount differences may be confused with differences due to morphology. Also, the sensitivity of mounts to orientation to wind or sun is nested within mount; without replication, inadvertent variation in mount positioning may be confused with differences among mounts. Data on heat loss from a of 1-day-old mallard duckling mount are used to illustrate orientation sensitivity.     

Seasonal changes and wind dependence of thermal conductance in dorsal fur from two small mammal species (Peromyscus leucopus and Microtus pennsylvanicus)

(1) We measured the thermal conductance of dorsal pelage from meadow voles (Microtus pennsylvanicus) and white-footed mice (Peromyscus leucopus) during summer and winter.

(2) Thermal conductance was lower in the winter pelage of both species, but the seasonal change was greater in meadow voles.

(3) The form of wind speed dependence was determined by fitting a nonlinear curve of the form a+bu^c to data recorded at five wind speeds. The most appropriate exponent c was between 0.908 and 0.987, depending on species and season. These values are common and suggest that thermal and dynamic forces are important.