Crystal structures of SCP2-thiolases of Trypanosomatidae, human pathogens causing widespread tropical diseases: the importance for catalysis of the cysteine of the unique HDCF loop

Thiolases are essential CoA-dependent enzymes in lipid metabolism. In the present study we report the crystal structures of trypanosomal and leishmanial SCP2 (sterol carrier protein, type-2)-thiolases. Trypanosomatidae cause various widespread devastating (sub)-tropical diseases, for which adequate treatment is lacking. The structures reveal the unique geometry of the active site of this poorly characterized subfamily of thiolases. The key catalytic residues of the classical thiolases are two cysteine residues, functioning as a nucleophile and an acid/base respectively. The latter cysteine residue is part of a CxG motif. Interestingly, this cysteine residue is not conserved in SCP2-thiolases. The structural comparisons now show that in SCP2-thiolases the catalytic acid/base is provided by the cysteine residue of the HDCF motif, which is unique for this thiolase subfamily. This HDCF cysteine residue is spatially equivalent to the CxG cysteine residue of classical thiolases. The HDCF cysteine residue is activated for acid/base catalysis by two main chain NH-atoms, instead of two water molecules, as present in the CxG active site. The structural results have been complemented with enzyme activity data, confirming the importance of the HDCF cysteine residue for catalysis. The data obtained suggest that these trypanosomatid SCP2-thiolases are biosynthetic thiolases. These findings provide promise for drug discovery as biosynthetic thiolases catalyse the first step of the sterol biosynthesis pathway that is essential in several of these parasites.

Photoacoustic tomography of the human finger: towards the assessment of inflammatory joint diseases

Inflammatory arthritis is often manifested in finger joints. The growth of new or withdrawal of old blood vessels can be a sensitive marker for these diseases. Photoacoustic (PA) imaging has great potential in this respect since it allows the sensitive and highly resolved visualization of blood. We systematically investigated PA imaging of finger vasculature in healthy volunteers using a newly developed PA tomographic system. We present the PA results which show excellent detail of the vasculature. Vessels with diameters ranging between 100 mu m and 1.5 mm are visible along with details of the skin, including the epidermis and the subpapillary plexus. The focus of all the studies is at the proximal and distal interphalangeal joints, and in the context of ultimately visualizing the inflamed synovial membrane in patients. This work is important in laying the foundation for detailed research into PA imaging of the phalangeal vasculature in patients suffering from rheumatoid arthritis.

Quantitative approaches for characterising fibrillar protein nanostructures

Polypeptide sequences have an inherent tendency to self-assemble into filamentous nanostructures commonly known as amyloid fibrils. Such self-assembly is used in nature to generate a variety of functional materials ranging from protective coatings in bacteria to catalytic scaffolds in mammals. The aberrant self-assembly of misfolded peptides and proteins is also, however, implicated in a range of disease states including neurodegenerative conditions such as Alzheimer's and Parkinson's diseases. It is increasingly evident that the intrinsic material properties of these structures are crucial for understanding the thermodynamics and kinetics of the pathological deposition of proteins, particularly as the mechanical fragmentation of aggregates enhances the rate of protein deposition by exposing new fibril ends which can promote further growth. We discuss here recent advances in physical techniques that are able to characterise the hierarchical self-assembly of misfolded protein molecnles and define their properties. © 2010 Materials Research Society.

Recognizing and managing common fish diseases

(4pp.)

A novel form of human disease with a protease-sensitive prion protein and heterozygosity methionine/valine at codon 129: Case report

Background: Sporadic Creutzfeldt-Jakob disease (sCJD) is a rare neurodegenerative disorder in humans included in the group of Transmissible Spongiform Encephalopathies or prion diseases. The vast majority of sCJD cases are molecularly classified according to the abnormal prion protein (PrPSc) conformations along with polymorphism of codon 129 of the PRNP gene. Recently, a novel human disease, termed "protease-sensitive prionopathy", has been described. This disease shows a distinct clinical and neuropathological phenotype and it is associated to an abnormal prion protein more sensitive to protease digestion. Case presentation: We report the case of a 75-year-old-man who developed a clinical course and presented pathologic lesions compatible with sporadic Creutzfeldt-Jakob disease, and biochemical findings reminiscent of "protease-sensitive prionopathy". Neuropathological examinations revealed spongiform change mainly affecting the cerebral cortex, putamen/globus pallidus and thalamus, accompanied by mild astrocytosis and microgliosis, with slight involvement of the cerebellum. Confluent vacuoles were absent. Diffuse synaptic PrP deposits in these regions were largely removed following proteinase treatment. PrP deposition, as revealed with 3F4 and 1E4 antibodies, was markedly sensitive to pre-treatment with proteinase K. Molecular analysis of PrPSc showed an abnormal prion protein more sensitive to proteinase K digestion, with a five-band pattern of 28, 24, 21, 19, and 16 kDa, and three aglycosylated isoforms of 19, 16 and 6 kDa. This PrPSc was estimated to be 80% susceptible to digestion while the pathogenic prion protein associated with classical forms of sporadic Creutzfeldt-Jakob disease were only 2% (type VV2) and 23% (type MM1) susceptible. No mutations in the PRNP gene were found and genotype for codon 129 was heterozygous methionine/valine. Conclusions: A novel form of human disease with abnormal prion protein sensitive to protease and MV at codon 129 was described. Although clinical signs were compatible with sporadic Creutzfeldt-Jakob disease, the molecular subtype with the abnormal prion protein isoforms showing enhanced protease sensitivity was reminiscent of the "protease-sensitive prionopathy". It remains to be established whether the differences found between the latter and this case are due to the polymorphism at codon 129. Different degrees of proteinase K susceptibility were easily determined with the chemical polymer detection system which could help to detect proteinase-susceptible pathologic prion protein in diseases other than the classical ones.

The economic impact of diseases and parasitic problems in freshwater fish production

Diseases and parasitic problems could constitute significant economic losses in fish production if not controlled, thus the need to continue monitoring its prevalence. Based on field studies on feral and intensively raised fish at the Kainji Lake Research Institute Nigeria, some diseases and parasitic problems have been identified. These include; helminthiasis; fungal disease; protozoa which include Myxosoma sp., Myxobolus spp., Henneguya sp., Trichodina sp., Ichthopthrius sp. bacterial mainly Aeromonas sp., Pseudomonas sp., mechanical injuries; death due to unknown causes and economic assessment of myxosporidian infection. Suggestion for disease control in fish production are recommended

Distribution and occurrence of bacterial fish diseases in the different cultural facilities of Lake Kainji area

Fish farming practices in the Lake Kainji Area of Nigeria are categorized under seven main cultural facilities, namely, earthen ponds/reservoirs, indoor/outdoor concrete tanks, plastic tanks, floating cages/hapas, aquaria, sewage and feral conditions. The presence of Bacteria isolates associated with diseased fish conditions varied significantly (P<0.05) with different cultural facilities. The highest bacteria isolates and bacterial disease incidence, 33% and 46% respectively, was associated with diseased fish in the indoor/outdoor concrete tanks. The least incidence of bacteria isolates (3.5%) and blue bacterial disease (3%) was associated with diseased fish in the aquaria and feral conditions. Nine Gram-negative and two Gram-positive bacteria genera were isolated during this investigation. Pseudomonas spp. (23.6%) and Staphylococcus spp. (14.3%), were the predominant Gram-negative and Gram-positive bacteria genera in the different cultural facilities, respectively. This paper highlights the relevance of occurrence and distribution of bacteria isolates associated with diseased fish to bacterial fish diseases under different cultural facilities

A review of diseases associated with cage culture systems: diagnosis and control in smale scale fish farming

Fish cage culture is a rapid aquacultural practice of producing fish with more yield compared to traditional pond culture. Several species cultured by this method include Cyprinus carpio, Orechromis niloticus, Sarotherodon galilaeus, Tilapia zilli, Clarias lazera, C. gariepinus, Heterobranchus bidorsalis, Citharinus citharus, Distochodus rostratus and Alestes dentes. However, the culture of fish in cages has some problems that are due to mechanical defects of the cage or diseases due to infection. The mechanical problems which may lead to clogged net, toxicity and easy access by predators depend on defects associated with various types of nets which include fold sieve cloth net, wire net, polypropylene net, nylon, galvanized and welded net. The diseases problems are of two types namely introduced diseases due to parasites. The introduced parasites include Crustaseans, Ergasilus sp. Argulus africana, and Lamprolegna sp, Helminth, Diplostomulum tregnna: Protozoan, Trichodina sp, Myxosoma sp, Myxobolus sp. the second disease problems are inherent diseases aggravated by the very rich nutrient environment in cages for rapid bacterial, saprophytic fungi, and phytoplanktonic bloom resulting in clogging of net, stagnation of water and low biological oxygen demand (BOD). The consequence is fish kill, prevalence of gill rot and dropsy conditions. Recommendations on routine cage hygiene, diagnosis and control procedures to reduce fish mortality are highlighted

Revealing, illuminating, and modifying proteins in human diseases using noncanonical amino acids

To better understand human diseases, much recent work has focused on proteins to either identify disease targets through proteomics or produce therapeutics via protein engineering. Noncanonical amino acids (ncAAs) are tools for altering the chemical and physical properties of proteins, providing a facile strategy not only to label proteins but also to engineer proteins with novel properties. My thesis research has focused on the development and applications of noncanonical amino acids in identifying, imaging, and engineering proteins for studying human diseases. Chapter 1 introduces the concept of ncAAs and reveals insights to how I chose my thesis projects.

ncAAs have been incorporated to tag and enrich newly synthesized proteins for mass spectrometry through a method termed BONCAT, or bioorthogonal noncanonical amino acid tagging. Chapter 2 describes the investigation of the proteomic response of human breast cancer cells to induced expression of tumor suppressor microRNA miR-126 by combining BONCAT with another proteomic method, SILAC or stable isotope labeling by amino acids in cell culture. This proteomic analysis led to the discovery of a direct target of miR-126, shedding new light on its role in suppressing cancer metastasis.

In addition to mass spectrometry, ncAAs can also be utilized to fluorescently label proteins. Chapter 3 details the synthesis of a set of cell-permeant cyclooctyne probes and demonstration of selective labeling of newly synthesized proteins in live mammalian cells using azidohomoalanine. Similar to live cell imaging, the ability to selectively label a particular cell type within a mixed cell population is important to interrogating many biological systems, such as tumor microenvironments. By taking advantage of the metabolic differences between cancer and normal cells, Chapter 5 discusses efforts to develop selective labeling of cancer cells using a glutamine analogue.

Furthermore, Chapter 4 describes the first demonstration of global replacement at polar amino acid positions and its application in developing an alternative PEGylation strategy for therapeutic proteins. Polar amino acids typically occupy solvent-exposed positions on the protein surface, and incorporation of noncanonical amino acids at these positions should allow easier modification and cause less perturbation compared to replacements at the interior positions of proteins.

On some new mathematical models for infective diseases: analysis, equilibrium, positivity and vaccination controls

196 p.

Combating fish disease. An explanation of the controls on notifiable fish diseases in Great Britain and advice on steps to prevent the spread of disease

For many years action has been taken to prevent the introduction and spread of serious fish diseases in Great Britain. In 1993 national rules were replaced by European Union wide rules designed to promote trade within the single market while safeguarding those parts of the Union with a high fish health status - such as this country. This booklet details the checks and controls which are applied to prevent the spread of disease outbreaks in this country. One can see that different rules apply to different diseases, generally reflecting the severity and other characteristics of the disease. The booklet also tries to explain the diseases and helps to recognise symptoms. This booklet is split into three parts: Part 1 gives an overview of the controls; Part 2 gives details for each of the diseases; and Part 3 gives advice on some of the precautions you can take to guard against the spread of disease.

Molecular probes and polymerase chain reaction (PCR) -based kits for diagnosis of shrimp diseases

Technology for effective and fast diagnosis of animal diseases is essential for developing aquaculture management strategies. This paper reviews the conventional techniques for shrimp disease diagnosis and discusses the emergence of nuclei acid probes and polymerase chain reaction (PCR)-based kits as powerful tools for rapid and accurate detection of shrimp diseases.

The potential for crop rotation in controlling diseases in shrimp culture

The use of antibiotics and other chemicals in controlling shrimp pathogens become ineffective as the strains grow more resistant to these chemicals. Moreover, the bacterial pathogen (Vibrio harveyi) produced biofilm coating that protects it from dying and disinfection procedures that are followed during pond preparation. Biological control is being considered as an alternative means of preventing shrimp disease outbreak. The main principle behind biological control is to enhance the growth of beneficial microorganisms which serve as antagonists or target pathogens. The paper discusses shrimp and tilapia crop rotation as a form of effective biological control, a technique which is already being practiced in Indonesia and the Philippines.