Teaching mothers to provide home treatment of malaria in Tigray, Ethiopia: a randomised trial.

BACKGROUND: No satisfactory strategy for reducing high child mortality from malaria has yet been established in tropical Africa. We compared the effect on under-5 mortality of teaching mothers to promptly provide antimalarials to their sick children at home, with the present community health worker approach. METHODS: Of 37 tabias (cluster of villages) in two districts with hyperendemic to holoendemic malaria, tabias reported to have the highest malaria morbidity were selected. A census was done which included a maternity history to determine under-5 mortality. Tabias (population 70,506) were paired according to under-5 mortality rates. One tabia from each pair was allocated by random number to an intervention group and the other was allocated to the control group. In the intervention tabias, mother coordinators were trained to teach other local mothers to recognise symptoms of malaria in their children and to promptly give chloroquine. In both intervention and control tabias, all births and deaths of under-5s were recorded monthly. FINDINGS: From January to December 1997, 190 of 6383 (29.8 per 1000) children under-5 died in the intervention tabias compared with 366 of 7294 (50.2 per 1000) in the control tabias. Under-5 mortality was reduced by 40% in the intervention localities (95% CI from 29.2-50.6; paired t test, p<0.003). For every third child who died, a structured verbal autopsy was undertaken to ascribe cause of mortality as consistent with malaria or possible malaria, or not consistent with malaria. Of the 190 verbal autopsies, 13 (19%) of 70 in the intervention tabias were consistent with possible malaria compared with 68 (57%) of 120 in the control tabias. INTERPRETATION: A major reduction in under-5 mortality can be achieved in holoendemic malaria areas through training local mother coordinators to teach mothers to give under-5 children antimalarial drugs.

X-ray scatter signatures for normal and neoplastic breast tissues.

Measurements of breast tissue scattering properties have been made in an energy dispersive x-ray diffraction system over the momentum transfer range of 0.70 to 3.50 nm(-1). One hundred samples of excised tissue have been used. Results from the diffraction system have been compared with the histological analysis for each individual sample. It has been found that tissue types can be characterized on the basis of the shape of the scatter spectrum and on its relative intensity. The shapes are significantly different between tissue types in the range 1.0 to 1.8 nm(-1) and suggest that if particular values of momentum transfer are monitored, a discriminating signal could be obtained. Analysis of the maximum intensity in the signature also reveals a change of up to a factor of 2 between adipose and fat-free tissues.

Temporal and spatial distribution of anopheline mosquitos in an Ethiopian village: implications for malaria control strategies.

The spatial and temporal distribution of Anopheles gambiae mosquitos in houses in the village of Sille in Ethiopia was monitored in 1990-91. Monthly mosquito densities in over 300 houses were obtained, and the data for each month were plotted on maps, which indicated clustering of mosquitos within the village. Spatial analysis using "kriging" techniques demonstrated clustering towards the edges of the village, the pattern of which changed with time. For example, the low density of mosquitos in one area in September increased as the nearby irrigation canals dried up during the following months. Since most entomological activity occurred at the periphery of the village, focal spraying of these areas could be a cost-effective procedure. If such clustering occurs in other villages, selective control of breeding sites and indoor spraying could provide a more efficient use of limited resources than traditional total coverage.

Prospects for the use of larvivorous fish for malaria control in Ethiopia: search for indigenous species and evaluation of their feeding capacity for mosquito larvae.

Because of problems with drug and insecticide resistance, the National Organization for the Control of Malaria and other Vectorborne Diseases, Ethiopia, has embarked on a programme of research on alternative malaria control methods, including the use of biological control agents, such as larvivorous fish. The objectives of the study were to identify indigenous larvivorous fish species which could be potential candidates for use as biological control agents; to extend knowledge of their distribution in Ethiopia; and to conduct laboratory tests to determine their feeding capacity. An extensive search resulted in the identification of 11 larvivorous fish species indigenous to Ethiopia, including five species previously unrecorded in the country. Seven species were assessed under standard laboratory conditions for their feeding capacity on larvae of Anopheles gambiae s.l. and Culex andersoni. All species tested were efficient larvivores in the laboratory. However, their larvivorous capacity should be tested further in field trials. Based on the findings of this study, two priority areas for the assessment of biological control using larvivorous fish were identified, the port city of Assab, using the local species Aphanius dispar, and the Ogaden, south-eastern Ethiopia, using the local species Oreochromis spilurus spilurus.

Cloning of developmentally regulated genes from Leishmania major and expression following heat induction.

The protozoa Leishmania undergo morphological and biochemical transformation from the promastigote to the amastigote form during their life cycle. To characterize this transformation process, we constructed a cDNA library for the promastigote stage of Leishmania major and used differential cDNA hybridization to identify cDNA sequences expressed at different abundance in promastigotes or amastigotes of L. major. P100/11E is a single copy gene whose 1600-nucleotide mRNA is enriched in promastigotes. P101/10 is a repeated gene whose 3300-nucleotide transcript is enriched in amastigotes. These developmentally regulated genes are not linked in the genome of L. major and are located on separate chromosome bands. The abundance of the P101/10 transcript increases severalfold during the transformation process at 37 degrees C in vitro and is thermally induced within 3 h after transfer of promastigotes from 27 to 37 degrees C. Examination of beta-tubulin gene expression showed that the relative abundance of the 3400-nucleotide beta-tubulin RNA is decreased at 37 degrees C in vitro. Our results indicate that the expression of two developmentally regulated genes of L. major is controlled at the level of mRNA abundance and provide direct evidence that thermal induction plays a general role in regulating gene expression in Leishmania.

Peculiar sequence organization of kinetoplast DNA minicircles from Trypanosoma cruzi.

The sequences of two minicircles from the kinetoplast DNA of the CL strain and one of the Y strain of Trypanosoma cruzi are reported. These 1.4 kb molecules have a peculiar sequence organization, the most distinctive feature being the occurrence of a 120 bp sequence repeated four times, located at 0, 90, 180 and 270 degrees along each circle. We have termed these conserved regions in this species 'minirepeats'. Minirepeats have a 3-fold higher concentration of cytosine residues in comparison with the variable regions and contain the universal 12-mer motif GGGGTTGGTGTA present in all sequenced minicircles and which was shown to be involved in DNA replication. A consensus sequence of T. cruzi minirepeats was determined using the 20 minirepeats present in five known T. cruzi minicircle sequences. This consensus sequence contains regions which have been remarkably well preserved in strains which show great biological diversity. In addition a low level of intraminicircle sequence similarity was also observed within the variable region, but this similarity did not extend between strains. The abundance of conserved minirepeat sequences containing invariant restriction sites in T. cruzi cells may prove valuable for the development of new direct diagnostic methods for Chagas' disease based on DNA probe technology.

Trehalose and maltose metabolism in yeast transformed by a MAL4 regulatory gene cloned from a constitutive donor strain.

A 6.8 kb fragment of DNA containing the regulatory sequence MAL4p has been cloned from a genomic library prepared from Saccharomyces cerevisiae strain 1403-7A which ferments maltose constitutively. The library was prepared by ligation of 5-20 kb Sau3AI restriction fragments of total yeast DNA into the BamH1 restriction site of shuttle vector YEp13. A restriction map of the cloned fragment indicates that it encompasses a 2.6 kb segment which closely resembles the regulatory MAL6 gene previously identified (Needleman et al. 1984). The hybrid plasmid, p(MAL4p)4, could transform maltose-nonfermenting strains which contain cryptic alpha-glucosidase and maltose permease genes (malp MALg), but could not transform strains containing a functional regulatory sequence and a defective maltase-permease region (MAlp malg). A correlated absence of maltase and permease DNA from the cloned fragment was indicated by the restriction map. Although the cloned DNA fragment was derived from a constitutive strain, maltose fermentation and alpha-glucosidase formation by yeast transformed with p(MAL4p)4 was largely inducible by maltose and sensitive to catabolite repression. Moreover, the active trehalose accumulation pattern (TAC(+) phenotype) linked to the complete MAL4 locus in strain 1403-7A and other constitutive MAL strains (Oliveira et al. 1981b) was not found in p(MAL4p)4 transformants. It may be concluded that constitutivity of maltose fermentation and the associated active trehalose accumulation are not merely consequences of a cis-dominant mutation causing constitutive formation of the MALp regulatory product. Moreover, constitutivity may not be caused solely by a mutation within the structural region of the MALp gene.

An Sau3 AI restriction endonuclease isoschizomer from Bacillus cereus.

The isolation and characterization of a restriction endonuclease from Bacillus cereus IOC 243 are described. The enzyme recognizes the palindromic sequence 5'-G(met-A,A)TC-3' as determined by PEI chromatography of pancreatic DNase, snake venom phosphodiesterase digestion products of labelled fragments, analysis of restriction digests from normal and N6-methyladenine-free DNA and direct sequence analysis of cloned fragments. The staggered cleavage products with 5' -terminal pGATC extensions are efficiently labelled with polynucleotide kinase and are easily cloned into BamHI sites. The enzyme, denoted Bce243, is thus an isoschizomer of Sau3AI. Its use and potential advantages in substituting Sau3AI. are discussed.

Sequence heterogeneity and anomalous electrophoretic mobility of kinetoplast minicircle DNA from Leishmania tarentolae.

Several unit-length minicircles from the kinetoplast DNA of Leishmania tarentolae were cloned into pBR322 and into M13 phage vectors. The complete nucleotide sequences of three different partially homologous minicircles were obtained. The molecules contained a region of approx. 80% sequence homology extending for 160-270 bp and a region unique to each minicircle . A 14-mer was found to be conserved in all kinetoplast minicircle sequences reported to date. The frequency distributions of various minicircle sequence classes in L. tarentolae were obtained by quantitative gel electrophoresis and by examination of the "T ladder" patterns of minicircles randomly cloned into M13 at several sites. By these methods we could assign approx. 50% of the total minicircle DNA into a minimum of five sequence classes. A sequence-dependent polyacrylamide gel migration abnormality was observed with several minicircle fragments both cloned and uncloned. The abnormality was dependent on the presence of a portion of the conserved region of the minicircle .

The kinetoplast DNA of the hemoflagellate protozoa.

The kinetoplast DNA of Leishmania tarentolae and Trypanosoma brucei was studied in terms of genetic organization and transcriptional activity. Several minor sequence classes of minicircles from L. tarentolae were cloned in a bacterial plasmid and were compared in terms of sequence organization. The cloned minicircles were characterized by the possession of a constant region of at least 91 nucleotides and a variable region. Minicircles from Trypanosoma brucei strain 366D were also cloned. Fragments of the maxicircle DNA from both species were also cloned in pBR322. No homology with the cloned minicircles was apparent. Several maxicircle transcripts, in addition to the 9 and 12s presumptive ribosomal RNAs, were observed in L. tarentolae. The 9 and 12s RNA genes were also mapped on the T. brucei maxicircle. Sequence homology between the L. tarentolae 9 and 12s RNA genes and the T. brucei 9 and 12s RNA genes was observed. A culture system was developed to study the developmental change of cultured bloodstream forms of T. brucei into procyclic forms. This developmental system is amenable for the study of the role of the kinetoplast DNA in the extensive mitochondrial biogenesis that occurs at this time.