Entomological investigations in the region of the last malaria focus in Morocco

To evaluate the risk of malaria transmission resumption in Morocco, we have studied the current level of receptivity of the region of the last malaria focus in the country. Anopheles (Anopheles) maculipennis labranchiae and Anopheles (Cellia) sergentii, the major vectors of malaria in Morocco, are still presents but their anthropothic index was low and no parasite positive samples were detected. An. labranchiae was very rare; only 34 females were caught over all the study period. The human biting rate was nil and none of its blood meal was human. An. sergenti was more abundant but its low human aggressiveness and its zoophilic behaviour would not attribute to this species an important vectorial capacity. Thus, the receptivity of Chefchaouen province, the region of the last malaria focus in Morocco, under the current vector control measures undertaken by Public Health services, is low and despite the likely presence of Plasmodium vivax gametocyte carriers, the malariogenic potential appears to be low and the risk of malaria resumption is, at this time, unimportant.     

Phylogeography of Barbary macaques (Macaca sylvanus) and the origin of the Gibraltar colony

The Barbary macaque (Macaca sylvanus) is the earliest offshoot of the genus Macaca and the only extant African representative, all other species being Asiatic. Once distributed throughout North Africa, M. sylvanus is now restricted to isolated forest fragments in Algeria and Morocco. The species is threatened; the maximum total wild population size is estimated at 10,000 individuals. Relationships among surviving wild subpopulations in Algeria (96 samples) and Morocco (116 samples) were examined by using 468-bp sequences from hypervariable region I of the mitochondrial DNA control region. Twenty-four different haplotypes were identified, differing by 1-26 mutational steps (0.2-5.6%) and 1 insertion. With one exception (attributable to secondary introduction in coastal Morocco), Algerian and Moroccan haplotypes are clearly distinct. However, whereas Moroccan subpopulations show little divergence in hypervariable region I sequences and little correspondence with geographical distribution, there is a deep division between two main subpopulations in Algeria and one marked secondary division, with haplotypes generally matching geographical distribution. Accepting an origin of the genus Macaca of 5.5 million years ago, the Moroccan population and the two main Algerian subpopulations diverged approximately 1.6 million years ago. Distinction between Moroccan and Algerian haplotypes permitted analysis of the origin of the Gibraltar colony of Barbary macaques (68 samples; 30% of the population). It is generally held that the present Gibraltar population descended from a dozen individuals imported during World War II. However, the Gibraltar sample was found to include Algerian and Moroccan haplotypes separated by at least 16 mutational steps, revealing a dual origin of the founding females.     

Reactive and undifferentiated arthritis in North Africa: use of PCR for detection of Chlamydia trachomatis

Little is known about the possible role of Chlamydia in patients with reactive or unclassified arthritis in North Africa. This study used polymerase chain reaction (PCR) to survey this population. In addition, we compared the results in three different laboratories for PCR analyses for Chlamydia trachomatis (Ct) in synovial fluid (SF) and tissue (ST) from these North African patients with reactive arthritis (ReA), undifferentiated arthritis (UA), and in rheumatoid arthritis (RA) and osteoarthritis (OA). Eight ReA (six posturethritic, two postenteritic), 23 UA, 13 OA, and 12 RA patients were studied in Algeria, Morocco, and Tunisia. Serum, SF, and ST were obtained from each patient. Ct-PCR was performed in the three different laboratories and compared to Ct-serology [microimmunofluorescence (MIF) and anti-hsp60 enzyme-linked immunosorbent assay (ELISA)] performed in one laboratory. The rate of Ct-PCR positivity in SF/ST was low: none out of the eight ReA and three out of 23 UA patients. In the controls, Ct DNA was detected in two OA SF and in one RA SF. There was no concordance for Ct-PCR positivity between the three laboratories. MIF suggested previous Ct infection (IgG-positive) in two out of five posturethritic ReA, none out of one postenteritic ReA, one out of 17 UA, and nine out of 21 RA/OA patients tested. No MIF-positive patient was PCR-positive from SF or ST. However, anti-hsp60 IgG was detected in all four out of four patients positive by PCR and in 11 out of 44 PCR-negative patients (p = 0.002). In this multinational comparative study, the rate of Ct-PCR-positive synovial specimens in North African ReA/UA patients was low. Concordance among the three PCR testing laboratories was poor indicating the need for test standardization. All Ct-PCR-positive patients were found positive by anti-hsp60 IgG serology. J. G. Kuipers and J. Sibilia contributed equally to this study. H. R. Schumacher and M. Dougados are equal senior authors of this study.     

Genetic analysis of ticks belonging to the Rhipicephalus sanguineus group in Latin America

Phylogenetic analyses based on mitochondrial 16S rDNA sequences were generated from Rhipicephalus sanguineus group specimens collected in 29 localities among 9 Latin-American countries, plus ticks collected in South Africa, Spain, and Italy. Sequences from Latin America generated six different haplotypes (A, B, C, D, E, and F). Phylogenetic analyses generated trees that segregated our tick sequences into two distinct clades: one is represented by haplotypes A–C, and South African R. sanguineus and Rhipicephalus turanicus ticks; the second clade is represented by haplotypes D–F, and European R. sanguineus and R. turanicus ticks. When haplotypes A–F are plotted in the Latin America map according to their geographical coordinates, it is clearly seen that haplotypes D–F are restricted to the southern portion of this continent, whereas haplotypes A–C are distributed in areas between northern Mexico and Brazil (except for the extreme south of this last country, where haplotype E was present). Hence, our phylogenetic analyses separated New World specimens of R. sanguineus into two distinct clades, one represented by tropical and subtropical populations (haplotypes A–C), here designated as the ‘tropical’ species. On the other hand, haplotypes D–F are here designated as the ‘temperate’ species because of their distribution in the southern portion of South America. Until recently, it was assumed that the R. sanguineus group was represented by a single species in the New World, namely R. sanguineus. While the present results coupled with recent studies support the presence of at least two species under the taxon R. sanguineus in the New World, they also show that even in the Old World, the taxon R. sanguineus might be represented by more than one species, since our phylogenetic analysis segregated European and South African R. sanguineus ticks into two distinct clades. The same can be applied for Spanish and South African R. turanicus.     

Dynamics of insecticide resistance in the malaria vector Anopheles gambiae s.l. from an area of extensive cotton cultivation in Northern Cameroon

Objective To explore temporal variation in insecticide susceptibility of Anopheles gambiae s.l. populations to the four chemical groups of insecticides used in public health and agriculture, in close match with the large‐scale cotton spraying programme implemented in the cotton‐growing area of North Cameroon. Methods Mosquito larvae were collected in 2005 before (mid June), during (mid August) and at the end (early October) of the cotton spraying programme. Larvae were sampled in breeding sites located within the cotton fields in Gaschiga and Pitoa, and in Garoua, an urban cotton‐free area that served as a control. Insecticide susceptibility tests were carried out with 4% DDT (organochlorine), 0.4% chlorpyrifos methyl (organophosphate), 0.1% propoxur (carbamate), 0.05% deltamethrin and 0.75% permethrin (pyrethroids). Results Throughout the survey, An. gambiae s.l. populations were completely susceptible to carbamate and organophosphate, whereas a significant decrease of susceptibility to organochlorine and pyrethroids was observed during spraying in cotton‐growing areas. Tolerance to these insecticides was associated with a slight increase of knockdown times compared to the reference strain. Among survivor mosquitoes, the East and West African Kdr mutations were detected only in two specimens of An. gambiae s.s. (n = 45) and not in Anopheles arabiensis (n = 150), suggesting metabolic‐based resistance mechanisms. Conclusions Environmental disturbance due to the use of insecticides in agriculture may provide local mosquito populations with the enzymatic arsenal selecting tolerance to insecticides.     

Alcohol-metabolizing enzyme gene polymorphisms in the Basque Country, Morocco, and Ecuador

Background: Genes ADH1B and ADH1C have certain functional SNPs that are related to alcoholism. The frequencies of these polymorphisms vary between populations, so studying them in populations made up of groups with different phylogeographic origins requires an individualized analysis of each group. In the Basque Country, various recently arrived foreign groups live side by side with the original Southern European population, particularly North Africans from Morocco and Hispanics from Ecuador. This study sets out to examine the distribution of the frequencies of alleles that encode alcohol dehydrogenase with different metabolization rates, as higher rates make for greater susceptibility to alcoholism. Methods: Four SNPs: rs1229984, rs2066702, rs698, and rs1693482 using Taqman technology with a Rt‐PCR were studied in a sample of 114 European individuals originating from the Basque Country, 100 North Africans from Morocco, and 109 Hispanics from Ecuador. The allele and genotype frequencies were calculated using Genepop v4.0. The most frequent haplotypes were estimated using the ELB algorithm with Arlequin v3.01. A breakdown of the complete disequilibrium commonly observed between the 2 missense polymorphisms that distinguish the common ADH1C alleles rs698 and rs1693482 was observed and confirmed by sequencing in 2 individuals from the Basque Country. Results: A higher frequency of protective allele ADH1C*1 was found in the North African population group. Haplotype combinations are also studied, and the rare association of alleles ADH1B*2–ADH1C*2 was observed in the Southern European group in the Basque Country, along with an allele not hitherto described in the ADH1C locus. Conclusions: This study provides the first data published on the allele and genotype frequencies of the ADH1C locus in the Moroccan population and on the ADH1B and ADH1C loci in the Ecuadorian population. The study shows differences in the distribution of the frequency of allele ADH1C*1 between the Basque Country and Moroccan populations, and a new allele not described to date.     

Natural history of Helicobacter pylori infection

This report describes the modalities of chronic gastritis induced by Helicobacter pylori infection in different populations. The full gamut of lesions representing the precancerous cascade is very prevalent in populations of low socioeconomic background experiencing very high gastric cancer risk, as seen in the Latin American Andes Mountains. In populations of high socioeconomic standards and high cancer risk, such as Japan and Korea, the precancerous cascade predominates and “early” cancers are also diagnosed frequently. Some reports describe frequent corpus atrophy, not prominent in the former group. The so-called African enigma is seen in populations of low socioeconomic standards, usually living at low altitudes, with high prevalence of infection but low frequency of cancer and precancerous lesions. In populations in transition from high to low cancer risk, duodenal ulcer and antral non-atrophic gastritis are frequently seen. In affluent societies at low risk of cancer, such as Western Europe, Australia and North America, mild non-atrophic gastritis associated with low virulence Helicobacter pylori genotypes predominate. The varied phenotypes of gastritis may reflect secular changes in the ecology of our species.     

North African Jewish and non-Jewish populations form distinctive, orthogonal clusters

North African Jews constitute the second largest Jewish Diaspora group. However, their relatedness to each other; to European, Middle Eastern, and other Jewish Diaspora groups; and to their former North African non-Jewish neighbors has not been well defined. Here, genome-wide analysis of five North African Jewish groups (Moroccan, Algerian, Tunisian, Djerban, and Libyan) and comparison with other Jewish and non-Jewish groups demonstrated distinctive North African Jewish population clusters with proximity to other Jewish populations and variable degrees of Middle Eastern, European, and North African admixture. Two major subgroups were identified by principal component, neighbor joining tree, and identity-by-descent analysis-Moroccan/Algerian and Djerban/Libyan-that varied in their degree of European admixture. These populations showed a high degree of endogamy and were part of a larger Ashkenazi and Sephardic Jewish group. By principal component analysis, these North African groups were orthogonal to contemporary populations from North and South Morocco, Western Sahara, Tunisia, Libya, and Egypt. Thus, this study is compatible with the history of North African Jews-founding during Classical Antiquity with proselytism of local populations, followed by genetic isolation with the rise of Christianity and then Islam, and admixture following the emigration of Sephardic Jews during the Inquisition.     

Species B of Anopheles culicifacies (Diptera: Culicidae) is reproductively less fit than species A and C of the complex

Anopheles culicifacies, the most important malaria vector of peninsular India exist as a complex of five sibling species. The member species of the complex have various biological differences including their susceptibility to malaria parasites. The present attempt is made to study and compare the fecundity of the differentially susceptible members of the An. culicifacies complex. Gravid female mosquitoes of species A, B and C were allowed to lay their eggs individually during first and second gonotrophic cycle. The eggs were counted after hatching and categorized as ‘hatched eggs’, ‘unhatched eggs’, ‘embryonated eggs’, ‘unembryonated eggs’ and ‘non/partially melanized eggs’. The data was analyzed using Student's t test, ANOVA, Chi-square and Pearson's correlation analysis. All females that were visually categorized as ‘gravid’ did not lay eggs. Species C laid maximum number of eggs per female. The eggs laid per female mosquito of each species were found to be significantly higher during second gonotrophic cycle as compared with the first gonotrophic cycle. The eggs hatched per female in species C were found to be significantly higher than that of species A and B. The poor-vector species B mosquitoes were found to be the least fecund among the members of the species complex. The unembryonated eggs constitute the biggest proportion of the unhatched eggs in species A, B and C of the species complex.     

Lower frequency of the 5/5 homozygous CNDP1 genotype in South Asian Surinamese

We investigated the frequency of the 5/5 homozygous CNDP1 (carnosinase) genotype, which was found to be associated with a reduced risk of developing diabetic nephropathy, in three ethnic groups in The Netherlands. Particularly interesting were the South Asian Surinamese, who have a high prevalence of diabetic nephropathy. Furthermore, we investigated the association between this gene and carnosinase activity in South Asian Surinamese and whether carnosinase was expressed in the kidney.We genotyped 290 South Asian Surinamese, 532 African Surinamese, and 472 White Dutch in a cross-sectional population study. Furthermore, an independent cohort of South Asian Surinamese was genotyped. In this population, carnosinase activity was measured in serum. Immunostaining and in situ hybridization for CNDP1 were performed on kidney tissue.Both South Asian populations had lower frequencies of the 5/5 homozygous genotype than African Surinamese and White Dutch (23.0%, 27.2%, 38.2%, and 41.3%, respectively; chi-square, p < 0.001). This genotype showed a lower carnosinase activity in South Asian Surinamese (Wilcoxon rank-sum, p = 0.03). CNDP1 was expressed in the kidney.South Asian Surinamese have a lower frequency of the 5/5 homozygous genotype, which was associated with lower carnosinase activity. Our study provides an indication that South Asian Surinamese are genetically at risk for developing diabetic nephropathy.     

Prevalence of HFE mutations in people from North Africa living in southern France

The two main mutations of the HFE (haemochromatosis) gene, C282Y and H63D, were found previously to be rare or absent among Africans. Dried blood samples of 1276 newborns from southern France were analysed for both HFE mutations, and the origins of the four grandparents of each newborn were recorded. The allele frequency of C282Y and H63D was 3.0% +/- 0.7% and 16.9% +/- 1.5% respectively. In a subgroup of 171 newborns with four North African ancestries (mainly from Morocco and Algeria) the allele frequency was 0.9%+2.5%-0.2% for the C282Y and 13.2% +/- 3.6% for H63D. HFE mutations are not absent in individuals with North African origins living in southern Europe. This finding has implications for the diagnosis and screening of hereditary haemochromatosis in these populations.     

First record of the Asian malaria vector Anopheles stephensi and its possible role in the resurgence of malaria in Djibouti,Horn of Africa

Anopheles stephensi is an important vector of urban malaria in India and the Persian Gulf area. Its previously known geographical range includes southern Asia and the Arab Peninsula. For the first time, we report A. stephensi from the African continent, based on collections made in Djibouti, on the Horn of Africa, where this species’ occurrence was linked to an unusual urban outbreak of Plasmodium falciparum malaria, with 1228 cases reported from February to May 2013, and a second, more severe epidemic that emerged in November 2013 and resulted in 2017 reported malaria cases between January and February 2014. Anopheles stephensi was initially identified using morphological identification keys, followed by sequencing of the Barcode cytochrome c-oxidase I (COI) gene and the rDNA second internal transcribed spacer (ITS2). Positive tests for P. falciparum circumsporozoite antigen in two of six female A. stephensi trapped in homes of malaria patients in March 2013 are evidence that autochthonous urban malaria transmission by A. stephensi has occurred. Concurrent with the second malaria outbreak, P. falciparum-positive A. stephensi females were detected in Djibouti City starting in November 2013. In sub-Saharan Africa, newly present A. stephensi may pose a significant future health threat because of this species’ high susceptibility to P. falciparum infection and its tolerance of urban habitats. This may lead to increased malaria outbreaks in African cities. Rapid interruption of the urban malaria transmission cycle, based on integrated vector surveillance and control programs aimed at the complete eradication of A. stephensi from the African continent, is strongly recommended.     

The First Case of Hb Groene Hart [α119(H2)Pro→Ser,CCT→TCT (α1)] Homozygosity Confirms That a Thalassemia Phenotype Is Associated with this Abnormal Hemoglobin Variant

Hb Groene Hart [α119(H2)Pro→Ser, CCT→TCT (α1)] has been reported in heterozygotes of Moroccan origin and also in association with the common ?α^3.7 deletion. In all cases, the mutated protein was not detectable but was apparently associated with a mild α-thalassemia (thal) phenotype, presumably due to a modification of the α-globin chain domain that is recognized by the a hemoglobin stabilizing protein (AHSP). The present case of Hb Groene Hart homozygosity, confirms that the α-thal phenotype is associated with this α-globin chain. Hb Groene Hart must be quite frequent not only in Morocco but probably also among the northern African coastal population.     

Taxonomic assessment of Anopheles crawfordi and An. dangi of the Hyrcanus Group of subgenus Anopheles in Vietnam

Anopheles dangi, introduced as a new species of the Hyrcanus Group of subgenus Anopheles in an illustrated dichotomous key for the identification of the Anopheles mosquitoes of Vietnam published in 1987, was distinguished from Anopheles crawfordi based on the presence of a humeral pale spot on the base of the costal vein of the wing. However, this character has been known to occur occasionally in An. crawfordi. To determine whether An. dangi is distinct from An. crawfordi, we analyzed nucleotide sequences of the COI, COII and Cyt-b genes of mtDNA and the D3 gene of rDNA obtained from specimens collected in south-central Vietnam that were identified as An. dangi and An. crawfordi based on the presence or absence, respectively, of a humeral pale spot. Maximum Likelihood and Bayesian analyses of the sequences showed a low mean genetic distance of 0.004 for specimens identified as An. crawfordi and 0.008 for those identified as An. dangi. The mean genetic distance between the two nominal species was 0.006, compared with 0.077 for any group versus the outgroup taxa Anopheles dirus and Anopheles minimus, and the specimens of the two forms clustered in a single strongly supported clade. Consequently, An. dangi is merely a morphological variant of An. crawfordi and is deemed to be a synonym of that nominal species.     

Investigations into the isolation of the Tukuyu focus of onchocerciasis (Tanzania) from S. damnosum s.l. vector re-invasion

As part of the feasibility study for an onchocerciasis vector elimination project we investigated the isolation of the Tukuyu focus in Tanzania from possible vector re-invasion. This was achieved by examining the distribution of the Simulium damnosum complex vector cytospecies outside the focus to look for potential sources of re-invasion. Besides cytotaxonomic identifications of the aquatic stages, we applied morphotaxonomic and molecular techniques to identify S. thyolense and confirm it as the anthropophilic species in both the Tukuyu and the neighbouring Ruvuma foci. We detected significant differences in chromosome inversion frequencies between the Tukuyu populations and those breeding to the southwest in the adjacent Songwe river basin and in northern Malawi (where there is no man-biting and no onchocerciasis), suggesting that there is not normally a great deal of migration in either direction. By contrast, populations of S. thyolense from the Tukuyu and Ruvuma foci (150 km southeast of Tukuyu) were much more similar in terms of their chromosomal polymorphisms, indicating a higher possibility of re-invasion, although migration is still restricted to some extent, as indicated by some differences in chromosome polymorphisms between the two foci. Future migratory events which might be associated with vector control operations can be monitored by vector cytospecies identification, the frequency of polymorphic inversions which characterise the different vector populations, and the identification of accompanying non-vector cytospecies (e.g. S. plumbeum and cytotype Kasyabone occur exclusively in the two foci, and hence their re-appearance in Tukuyu could have only one outside source). The morphology of the scutal pattern of neonate males may act as a quick test for vector species identification where chromosome squashes are unavailable.     

Larvicidal activity of synthesized silver nanoparticles using Eclipta prostrata leaf extract against filariasis and malaria vectors

Mosquitoes transmit serious human diseases, causing millions of deaths every year. Use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides of synthesized natural products for vector control have been a priority in this area. In this study, larvicidal activity of synthesized silver nanoparticles (AgNPs) utilizing aqueous extract from Eclipta prostrata, a member of the Asteraceae was investigated against fourth instar larvae of filariasis vector, Culex quinquefasciatus say and malaria vector, Anopheles subpictus Grassi (Diptera: Culicidae). The synthesized AgNPs characterized by UV–vis spectrum, scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared (FTIR) and X-ray diffraction (XRD). SEM analyses of the synthesized AgNPs were clearly distinguishable measured 35–60 nm in size. Larvae were exposed to varying concentrations of aqueous extract of synthesized AgNPs for 24 h. The maximum efficacy was observed in crude aqueous, and synthesized AgNPs against C. quinquefasciatus (LC₅₀ = 27.49 and 4.56 mg/L; LC₉₀ = 70.38 and 13.14 mg/L), and against A. subpictus (LC₅₀ = 27.85 and 5.14 mg/L; LC₉₀ = 71.45 and 25.68 mg/L) respectively. The chi-square value were significant at p < 0.05 level. These results suggest that the synthesized AgNPs have the potential to be used as an ideal eco-friendly approach for the control of the Culex tritaeniorhynchus and A. subpictus. This method is considered as a new approach to control vectors. Therefore, this study provides first report on the mosquito larvicidal activity of synthesized AgNPs against vectors.     

<Superscript>13</Superscript>C-Urea Breath Test for Success of <Emphasis Type="Italic">Helicobacter pylori</Emphasis> Eradication: Study of 5885 Israeli Patients.

Helicobacter pylori (Hp) infection is highly prevalent in many countries and may cause gastritis, peptic ulcer disease, gastric cancer, and lymphoma. Successful eradication depends on the specific treatment used, patient compliance, and Hp antibiotic resistance. The primary aim was to characterize groups of patients with one or more failures of Hp eradication treatment. The secondary aim was to evaluate the factors that influence eradication failure. Between April 1, 1998, and December 31, 2001, 5885 patients were studied for the success of Hp eradication with the ¹³C-urea breath test (¹³C-UBT): 5442 after one course of treatment (Group I), 380 after two courses (Group II), and 63 after three courses (Group III). The ¹³C-UBT was positive in 27.8%, 37.4%, and 47.6% of patients in Groups I, II, and III, respectively (P_I-II = 0.000, P_II-III = 0.126). A combination of omeprazole, amoxicillin, and clarithromycin (OAC) was used in 31.3%, 27.4%, and 7.9% of Groups I, II, and III, respectively, and a combination of omeprazole, amoxicillin, and metronidazole (OAM) in 15.2%, 28.9%, and 28.6%, respectively. Regimens that contained clarithromycin were used in decreasing order in Groups I, II, and III, and regimens containing metronidazole, bismuth, or tetracycline, in increasing order. The only good prognostic factor for successful eradication was Israeli origin, while European–American and Asian–African origin, recurrence of symptoms, a history of duodenal ulcer, and chronic proton pump inhibitor (PPI) use did not favor successful eradication. Our results suggest that origin, history of peptic disease, and chronic PPI use are predictors of eradication failure.     

Intraspecific variability (rDNA ITS and mtDNA Cyt b) of Phlebotomus sergenti in Spain and Morocco

Phlebotomus sergenti, the main vector of Leishmania tropica, is widely represented in Spain, whilst L. tropica is not an endemic species in this country. Nevertheless, the important human migrant flow from regions where L. tropica is endemic, the existence of its vector and the anthroponotic cycle of the parasite could lead to its establishment in our country. The vectorial capacity of P. sergenti could depend on the existence of cryptic species or races, which can only be identified by molecular methods. Our aim has been to study the molecular variation on two genes of Spanish populations of P. sergenti, comparing them with Moroccan ones. We have identified 5 ribosomal and 16 mitochondrial haplotypes. The results obtained indicate a high diversity of P. sergenti in Spain and the existence in the country of two P. sergenti lineages, a typically Spanish mitochondrial lineage and another one common in Morocco, where L. tropica is endemic in the south and emergent in the north and centre of the country.     

Effect of temperature and inter-specific competition on the development and survival of Anopheles gambiae sensu stricto and An. arabiensis larvae

The two major African malaria vectors Anopheles gambiae sensu stricto and An. arabiensis are sibling species that occupy different climatic niches but are frequently found in the same larval habitats. Differences in survival and development of the aquatic larval stages of these species at different temperatures may help explain adult distribution. The development time from first instar larva to adult at constant water temperatures (25, 30 and 35 °C) was measured in these two species when reared together in the same container (ratio 1:1) and separately. Survival to adult was highest in both species reared at 25 °C and decreased with increasing temperature. More adult An. gambiae s.s. were produced at 25 °C than An. arabiensis (80% interquartile range (78-88) versus 68% (63-78)) but this situation was reversed at 35 °C (7% (3-17) versus 33% (27-32)). The survival of An. gambiae s.s. when reared alone was similar to that when reared in the presence of An. arabiensis. In marked contrast An. arabiensis suffered reduced survival when raised with An. gambiae s.s. at 30 °C (20% (7-57)) than when reared independently (57% (45-72)). Mean age at eclosion and adult size decreased for both species with increasing water temperature, however An. arabiensis larvae developed at a slower rate and resulted in larger adults than An. gambiae s.s. throughout. The apparent greater production of An. arabiensis at high water temperatures and An. gambiae s.s. at lower water temperatures may in part explain the spatial and temporal distribution of the two species.     

Analysis of SMAD4/DPC4 gene alterations in multiploid colorectal carcinomas

Background Although recent animal studies have shown that SMAD4/DPC4 gene alterations are essential for late-stage intestinal tumorigenesis, the role of SMAD4/DPC4 gene alterations in primary human colorectal carcinomas is not fully understood. Therefore, we attempted to clarify the role of the SMAD4/DPC4 gene during tumor progression of colorectal carcinoma.Methods Differences in allelic imbalance (AI) and mutations of the SMAD4/DPC4 gene between diploid and aneuploid populations were analyzed for 30 sporadic DNA multiploid colorectal carcinomas (used as a tumor progression model and defined as the coexistence of diploid and aneuploid cells within the same tumor). The crypt isolation technique was coupled with DNA cytometric sorting and a polymerase chain reaction assay. In addition, hypermethylation of the promoter region was examined to clarify whether inactivation of gene expression occurred.Results Although a SMAD4/DPC4 gene AI was detected in only 5 of 27 informative diploid populations, 25 of 27 aneuploid populations had a SMAD4/DPC4 gene AI. Mutation of the SMAD4/DPC4 gene was detected in only one aneuploid population of multiploid colorectal carcinomas, but not in the corresponding diploid population. In total, 20 available multiploid carcinomas were selected for methylation analysis, and no evidence of hypermethylation of the promoter region was found.Conclusions We suggest that, although mutation of the SMAD4/DPC4 gene and hypermethylation of the promoter region are infrequent events in colorectal tumorigenesis, AI at the SMAD4/DPC4 gene locus may play a key role in the progression of colorectal carcinomas.