Characterization of the DNF15S2 locus on human chromosome 3: identification of a gene coding for four kringle domains with homology to hepatocyte growth factor.

A human genomic DNA library was screened by using conditions of reduced stringency with a bovine cDNA probe coding for the kringle domains in prothrombin in order to isolate the human prothrombin gene. Twelve positives were identified, three of which coded for prothrombin (Degen & Davie, 1987). Phage L5 was characterized in more detail because of its strong hybridization to the cDNA probe and its unique restriction map compared to the gene coding for human prothrombin. The gene in L5 was sequenced and found to code for a kringle-containing protein. A human liver cDNA library was screened by using a genomic probe from the gene in L5. cDNAs were isolated that contained sequence identical with regions in the gene in L5. Comparison of the cDNA with the gene indicated that the gene in L5 was composed of 18 exons separated by 17 intervening sequences and is 4690 bp in length. Exons ranged in size from 36 to 242 bp in length while intervening sequences ranged from 77 to 697 bp in length. The putative protein encoded by the gene in L5 contains four kringle domains followed by a serine protease-like domain. This domain structure is identical with that found in hepatocyte growth factor (HGF), although the two proteins are only about 50% identical. On the basis of the similarity of the protein encoded by L5 and HGF, we propose that the putative L5 protein be tentatively called HGF-like protein until a function is identified. The DNA sequence of the gene and cDNA and its translated amino acid sequence were compared against GenBank and NBRF databases. Sequences homologous to DNF15S1 and DNF15S2, human DNF15S2 lung mRNA, and rat acyl-peptide hydrolase were identified in exon 17 to the 3' end of the characterized sequence for the gene. From our results, it is apparent that the gene coding for human HGF-like protein is located at the DNF15S2 locus on human chromosome 3 (3p21). The gene for acyl-peptide hydrolase is 444 bp downstream of the gene coding for HGF-like protein, but on the complementary strand. The DNF15S2 locus has been proposed to code for one or more tumor suppressor genes since this locus is deleted in DNA from small cell lung carcinoma, other lung cancers, renal cell carcinoma, and von Hippel-Lindau syndrome.     

Permethrin-impregnated bednet effects on resting and feeding behaviour of lymphatic filariasis vector mosquitoes in Kenya

The impact of permethrin-impregnated bednets on resting and feeding behaviour of mosquito vectors of Wuchereria bancrofti, causing human lymphatic filariasis was studied in six pairs of villages (treated and untreated) before and after intervention. The study villages were in Kwale District, near the coast of Kenya, where Bancroftian filariasis is highly endemic, transmitted by a combination of both anopheline and culicine mosquito vectors. Mosquitoes were collected weekly in each village, indoors (using pyrethrum spray catches) and outdoors (using pit traps) during 3–4 months following the long rainy season. Of the filariasis vector species of mosquitoes collected in 1994 before intervention, 33.6% were members of the Anopheles gambiae complex, 30% were An. funestus and 36.4% were Culex quinquefasciatus. PCR analysis of the An. gambiae complex species collected in 1995 demonstrated that 98.5% were An. gambiae sensu stricto, 1% An. arabiensis and 0.5% An. merus. Introduction of impregnated bednets in 1995 significantly reduced the number of indoor-resting An. gambiae s.l. by 94.6% and An. funestus by 96.7%, but there was no change in the number of Cx quinquefasciatus collected indoors. The number of outdoor-resting An. gambiae s.l. was significantly reduced, whereas densities of An. funestus and Cx quinquefasciatus remained unaffected outdoors. ELISA analysis of mosquito bloodmeals showed a shift from human to animal feeding after the introduction of treated nets. The human blood index (HBI) for indoor resting Cx quinquefasciatus was reduced from 93.1% to 14.4%. Vector potential based on the HBI and mosquito density was estimated to be reduced by 99% for An. gambiae s.l., 98% for An. funestus and 97% for Cx quinquefasciatus and vectorial capacity would be suppressed even more by the impact on the vector survival rates (not measured). These results suggest that permethrin-impregnated bednets give effective personal protection against transmission of W. bancrofti by An. gambiae, An. funestus and Cx quinquefasciatus in East Africa.     

SGS--Spatial Genetic Software: a computer program for analysis of spatial genetic and phenotypic structures of individuals and populations.

We have developed a program called Spatial Genetic Software (SGS), which provides a user-friendly Windows tool to analyze both local and broad scale genetic and phenotypic structure. It can deal with nearly any type of genetic data, codominant (allozyme, PCR-RFLP, microsatellite) or dominant (RAPD, AFLP) markers, or biparentally (nuclear) or uniparentally (cpDNA and mtDNA) inherited markers. Data based on any of these markers can be analyzed, either as individual genotypes within a single population (local scale) or as allele or haplotype frequencies from different populations (broad scale). We also include a simple approach to analysis of spatial structure for continuous quantitative traits. The program implements various parameters to analyze spatial genetic and phenotypic structure: Moran's index, Geary's index, number of alleles in common, and approaches using genetic distances and F(ST) values. The statistical significance of all measures is verified by the use of a permutation test. The results are assessed by graphics that can be integrated, via the clipboard, to other Windows programs. The details of the computations are given in a table and can be stored as ASCII files.     

Relationship between host diversity and parasite diversity: flea assemblages on small mammals

Aim We examined the relationship between host species richness and parasite species richness using simultaneously collected data on small mammals (Insectivora, Rodentia and Lagomorpha) and their flea parasites. Location The study used previously published data on small mammals and their fleas from 37 different regions. All the world's main geographical regions other than Australasia and Wallacea were represented in the study, i.e. neotropical, nearctic, palaearctic, oriental and afrotropical realms. Methods We controlled the data for the area sampled and sampling effort and then tested this relationship using both cross‐region conventional analysis and the independent contrasts method (to control for the effects of biogeographic historical relationships among different regions). Brooks parsimony analysis was used to construct a region cladogram based on the presence/absence of a host species and host phylogeny. Results Both cross‐region and independent contrasts analyses showed a positive correlation between host species richness and flea species richness. Conventional cross‐region regression under‐ or overestimated fleas species richness in the majority of regions. Main conclusions When the regression derived by the independent contrasts method was mapped onto the original tip data space, points that deviated significantly from the regression originated from Kenya, Mississippi and southern California (lower than expected flea richness) and Chile, Idaho, south‐western California and Kyrgyzstan (higher than expected flea richness). These deviations can be explained by the environmental mediation of host–flea relationships and by a degree of environmental variety in sampled areas.     

Role of olivary electrical coupling in cerebellar motor learning

The level of electrotonic coupling in the inferior olive is extremely high, but its functional role in cerebellar motor control remains elusive. Here, we subjected mice that lack olivary coupling to paradigms that require learning-dependent timing. Cx36-deficient mice showed impaired timing of both locomotion and eye-blink responses that were conditioned to a tone. The latencies of their olivary spike activities in response to the unconditioned stimulus were significantly more variable than those in wild-types. Whole-cell recordings of olivary neurons in vivo showed that these differences in spike timing result at least in part from altered interactions with their subthreshold oscillations. These results, combined with analyses of olivary activities in computer simulations at both the cellular and systems level, suggest that electrotonic coupling among olivary neurons by gap junctions is essential for proper timing of their action potentials and thereby for learning-dependent timing in cerebellar motor control.     

Occurrence and direct control potential of parasitoids and predators of the fall armyworm (Lepidoptera: Noctuidae) on maize in the subtropical lowlands of Mexico

Abstract 1 Native natural enemies have the potential to control fall armyworm Spodoptera frugiperda (Smith) in tropical maize grown in Mexico, where this insect pest causes severe economic losses to farmers. It has been proposed that enhancing herbivore‐induced volatile emissions in maize plants may help to increase the effectiveness of natural enemies, which use these volatiles to locate their prey. This will only be of immediate benefit to farmers if the activity of the natural enemies results in a direct reduction in herbivory. Here we report on field surveys for the most common natural enemies in a tropical maize‐growing region in Mexico and the potential effects of these enemies on herbivory by fall armyworm. 2 Caterpillars were collected in maize fields near Poza Rica in the state of Veracruz during January and February 1999, 2000 and 2001. Plants were either naturally infested by S. frugiperda, or artificially infested with laboratory‐reared larvae. Ten species of parasitoids emerged from the collected larvae and eight species of predators that are known to feed on larvae and eggs were observed on the plants. Campoletis sonorensis (Cameron) (Hymenoptera: Ichneumonidae) was the dominant parasitoid species, in 1999 and 2001. 3 Of the nine larval parasitoids collected, six (all solitary) are known to reduce herbivory, whereas one causes the host to eat more (for two species this is not known). This implies that enhancing the effectiveness of solitary endoparasitoids may benefit subsistence farmers in developing countries by immediately reducing herbivory. The overall benefit for the plant resulting from parasitoid activity also has important implications for the evolutionary role of parasitoids in contributing to selection pressures that shape indirect defences in plants.     

Functional overlap between two classes of matrix-degrading proteases in wound healing.

Retarded wound healing was found in mice deficient in the serine protease precursor plasminogen, as well as in wild-type mice treated with the metalloprotease inhibitor galardin, but in both cases wound closure was ultimately completed in all mice within 60 days. The expression of several matrix metalloproteases in keratinocytes migrating to cover the wound was strongly enhanced by galardin treatment. However, when plasminogen-deficient mice were treated with galardin, healing was completely arrested and wound closure was not seen during an observation period of 100 days, demonstrating that protease activity is essential for skin wound healing. The requirement for both plasminogen deficiency and metalloprotease inhibition for complete inhibition of the healing process indicates that there is a functional overlap between the two classes of matrix-degrading proteases, probably in the dissection of the fibrin-rich provisional matrix by migrating keratinocytes. Each class alone is capable of maintaining sufficient keratinocyte migration to regenerate the epidermal surface, although this function would normally be performed by both classes acting in parallel. Since there are strong similarities between the proteolytic mechanisms in wound healing and cancer invasion, these results predict that complete arrest of this latter process in therapeutic settings will require the use of inhibitors of both classes of proteases.