Development of Fungal Applications on Netting Substrates for Malaria Vector Control

Mosquito resistance to chemical insecticides is considered a serious threat for the sustainable use of contemporary malaria vector control methods. Fungal entomopathogens show potential as alternative biological control agents against (insecticide-resistant) anophelines. This study was designed to test whether the fungus, Beauveria bassiana, could be delivered to mosquitoes on netting materials that might be used in house screens, such as eave curtains. Tests were conducted to determine effects of formulation, application method, netting material, and nature of mosquito contact. Beauveria had a twice as high impact on Anopheles gambiae s.s. longevity when suspended in Shellsol solvent compared with Ondina oil (HR = 2.12, 95% confidence interval = 1.83–2.60, P 90% of mosquitoes within 10 d. Results indicate that the use of fungal spores dissolved in Shellsol and sprayed on small-meshed cotton eave curtain nets would be the most promising option for field implementation. Biological control with fungus-impregnated eave curtains could provide a means to target host-seeking mosquitoes upon house entry, and has potential for use in integrated vector management strategies, in combination with chemical vector control measures, to supplement malaria control in areas with high levels of insecticide resistance

Bone Quality in Chronic Kidney Disease: Definitions and Diagnostics

PURPOSE OF REVIEW: In this paper, we review the epidemiology, diagnosis, and pathogenesis of fractures and renal osteodystrophy. RECENT FINDINGS: The role of bone quality in the pathogenesis of fracture susceptibility in chronic kidney disease (CKD) is beginning to be elucidated. Bone quality refers to bone material properties, such as cortical and trabecular microarchitecture, mineralization, turnover, microdamage, and collagen content and structure. Recent data has added to our understanding of the effects of CKD on alterations to bone quality, emerging data on the role of abnormal collagen structure on bone strength, the potential of non-invasive methods to inform our knowledge of bone quality, and how we can use these methods to inform strategies that protect against bone loss and fractures. However, more prospective data is required. CKD is associated with abnormal bone quality and strength which results in high fracture incidence

Tick saliva is a potent inhibitor of endothelial cell proliferation and angiogenesis

We report for the first time that saliva of the hard tick and Lyme disease vector,Ixodes scapularis,is a potent inhibitor of angiogenesis. Saliva (≤ 1:500 dilutions) or salivary gland (0.1–0.5 pairs/assay) dose-dependently inhibits microvascular endothelial cell (MVEC) proliferation. Inhibition was also detected with the saliva of the cattle tick Boophilus microplus but not with the salivary gland of Anopheles gambiae, An. stephensi, Lutzomyia longipalpis, Phlebotomus papatasi, Aedes aegypti, Culex quinquefasciatus, and Cimex lectularius. Inhibition of MVEC proliferation by I. scapularis saliva was accompanied by a change in cell shape (shrinkage of the cytoplasm with loss of cell-cell interactions) and apoptosis which was estimated by expression of phosphatidylserine using the Apopercentage dye, and by a typical pattern of chromatin margination, condensation, and fragmentation as revealed by nuclear staining with Hoechst 33258.The effect of saliva appears to be mediated by endothelial cell α5β1 integrin, because monoclonal antibodies against this but not αvβ3, αvβ5, α9β1, or α2β1 integrins remarkably block its effect. In addition, SDS/PAGE shows that saliva specifically degrades purified α5β1 but not αvβ5 or αvβ3 integrins. Incubation of saliva with EDTA and 1,10-phenanthroline, but not phenylmethylsulfonyl fluoride (PMSF), inhibits saliva-dependent degradation of purified α5β1 integrin, suggesting that a metalloprotease is responsible for the activity. Finally, saliva at ≤ 1:1,000 dilutions blocks sprouting formation from chick embryo aorta implanted in Matrigel, an in vitro model of angiogenesis. These findings introduce the concept that tick saliva is a negative modulator of angiogenesis-dependent wound healing and tissue repair, therefore allowing ticks to feed for days. Inhibition of angiogenesis was hitherto an unidentified biologic property of the saliva of any blood-sucking arthropod studied so far. Its presence in tick saliva may be regarded as an additional source of angiogenesis inhibitors with potential applications for the study of both vector and vascular biology

Cloning of a salivary gland metalloprotease and characterization of gelatinase and fibrin(ogen)lytic activities in the saliva of the Lyme disease tick vector Ixodes scapularis

The full-length sequence of tick salivary gland cDNA coding for a protein similar to metalloproteases (MP) of the reprolysin family is reported. The Ixodes scapularis MP is a 488 amino acid (aa) protein containing pre- and pro-enzyme domains, the zinc-binding motif HExxHxxGxxH common to metalloproteases, and a cysteine-rich region. In addition, the predicted amino-terminal sequences of I. scapularis MPs were found by Edman degradation of PVDF-transferred SDS/PAGE-separated tick saliva proteins, indicating that these putative enzymes are secreted. Furthermore, saliva has a metal-dependent proteolytic activity towards gelatin, fibrin(ogen), and fibronectin, but not collagen or laminin. Accordingly, I. scapularis saliva has a rather specific metalloprotease similar to the hemorrhagic proteases of snake venoms. This is the first description of such activity in tick saliva and its role in tick feeding and Borrelia transmission is discussed

Cloning and expression of hmgr1 gene from Hevea brasiliensis

Biosynthesis of natural rubber (cis-1,4-polyisoprene) takes place through mevalonate pathway in Hevea. The enzyme 3-hydroxy- 3-methyl glutaryl-CoA reductase (HMGR), which catalyses the synthesis of mevalonate from HMG-CoA is a key regulatory enzyme in this pathway. This study aimed to clone and express hmgr1 gene, in order to obtain the HMGR protein in vitro and to further use this protein as a marker for yield potential in Hevea. For this purpose, mRNA was isolated from the latex of Hevea (clone RRII 105). cDNA was synthesized and PCR amplification of coding region of hmgr1 was performed using hmgr1 specific primers. The PCR amplified product (~1.8 kb) was cloned into an expression vector (pRSET-A) and transformed into E. coli (BL21DE3) cells. Protein expression in transformed cells when monitored by SDS-PAGE analysis indicated the presence of HMGR protein (61.6 kDa). The protein would be used for developing specific antibody that could be further utilized for the quantification of HMGR in different Hevea clones for screening the yield potential. The details of cloning and expression of hmgr1 are presented and discussed

Nanostructure-modulated planar high spectral resolution spectro-polarimeter

We present a planar spectro-polarimeter based on Fabry-P{\'e}rot cavities with embedded polarization-sensitive high-index nanostructures. A $7~\mu$m-thick spectro-polarimetric system for 3 spectral bands and 2 linear polarization states is experimentally demonstrated. Furthermore, an optimal design is theoretically proposed, estimating that a system with a bandwidth of 127~nm and a spectral resolution of 1~nm is able to reconstruct the first three Stokes parameters \textcolor{black}{with a signal-to-noise ratio of -13.14~dB with respect to the the shot noise limited SNR}. The pixelated spectro-polarimetric system can be directly integrated on a sensor, thus enabling applicability in a variety of miniaturized optical devices, including but not limited to satellites for Earth observation

Purification of a serine protease and evidence for a protein C activator from the saliva of the tick, \u3cem\u3eIxodes scapularis\u3c/em\u3e

The saliva of ticks is critical to their survival as parasites and hematophagous animals. In this study, we have purified an enzyme with trypsin-like activity from the saliva of the tick vector of Lyme Disease, Ixodes scapularis. This enzyme, named as IXOSP (I. scapularis salivary serine protease), is a 29.9 kDa molecule with N-terminus FPxMVxLRIKxR. A BLAST search identified IXOSP as a secreted serine protease (AAY66740) with a conserved catalytic triad His, Asp, and Ser. In vitro studies demonstrated that IXOSP cleaves chromogenic substrates with arginine in the P1 position, by a mechanism inhibited by PMSF or aprotinin. Gene expression studies revealed that IXOSP is expressed at different tick developmental stages, including eggs, and unfed or fed adult tick salivary glands, but not in nymphs or in the midgut. While the physiological substrate for IXOSP remains to be identified, we demonstrated that I. scapularis saliva activate protein C (PC) resulting in the production of activated PC, a potent anticoagulant that also regulates a myriad of inflammatory responses through protease activated receptors. In contrast, the salivary glands of Anopheles gambiae, Anopheles stephensi, Anopheles albimanus, Aedes aegypti, Lutzomyia longipalpis, and Phlebotomus ariasi did not activate protein C. These discoveries are discussed in the context of blood coagulation, inflammation and vector–host interactions

Application of regulatory sequence analysis and metabolic network analysis to the interpretation of gene expression data

We present two complementary approaches for the interpretation of clusters of co-regulated genes, such as those obtained from DNA chips and related methods. Starting from a cluster of genes with similar expression profiles, two basic questions can be asked: 1. Which mechanism is responsible for the coordinated transcriptional response of the genes? This question is approached by extracting motifs that are shared between the upstream sequences of these genes. The motifs extracted are putative cis-acting regulatory elements. 2. What is the physiological meaning for the cell to express together these genes? One way to answer the question is to search for potential metabolic pathways that could be catalyzed by the products of the genes. This can be done by selecting the genes from the cluster that code for enzymes, and trying to assemble the catalyzed reactions to form metabolic pathways. We present tools to answer these two questions, and we illustrate their use with selected examples in the yeast Saccharomyces cerevisiae. The tools are available on the web (http://ucmb.ulb.ac.be/bioinformatics/rsa-tools/; http://www.ebi.ac.uk/research/pfbp/; http://www.soi.city.ac.uk/~msch/)