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Biomedical subjects

P D Ready

Publications and source records attributed to P D Ready.

At least 37 records · Page 2Linked to original sources
PubMed1995

A single isoform of the Na+/K(+)-ATPase alpha-subunit in Diptera: evidence from characterization of the first extracellular domain.

The first extracellular domain of the alpha-subunit of the Na+/K+-ATPase (sodium/potassium pump) is functionally important, affecting sensitivity of the enzyme to cardiac glycosides (e.g. ouabain) and being implicated in the transport of K+. This domain is also variable among mammalian isoforms of the alpha-subunit. Using PCR, we have isolated from seven insect species with contrasting physiologies a DNA fragment containing this region, in order to help determine whether tissue-specific expression might be associated with isoforms encoded by a gene family, as it is in mammals. A single sequence (with one ORF) characteristic of Na+/K+-ATPase was obtained from genomic DNA of each species. Only the fragment from Manduca sexta contained an intron, but at a location different to that found in mammals. For all Diptera so far characterized, the species phylogeny is the same as the alpha-subunit gene phylogeny (based on the sequences of the first extracellular domain and flanking transmembrane domains). The results strongly indicate a single, ouabain-sensitive isoform of the alpha-subunit of Na+/K+-ATPase is present in Diptera.

Amino Acid Sequence↗
PubMed1994

Isolation of non-LTR retrotransposon reverse transcriptase-like sequences from phlebotomine sandflies.

Reverse transcriptase-like sequences (RTs) with amino acid motifs characteristic of non-LTR retrotransposons have been isolated from several medically important phlebotomine sandfly species. These sequences were amplified using the polymerase chain reaction (PCR) with primers based on conserved amino acid motifs present in previously described insect RTs. A further set of RTs were amplified using primers based on the conserved regions identified in phlebotomine RTs. The average similarity of the phlebotomine RTs to the Drosophila I, F and R1Dm elements was 28-29% between the closest primers used. Phlebotomine RTs were 31-62% similar to each other, the most dissimilar sequences coming from the same species. Several amino acid residues were invariant in the ten phlebotomine RTs, including motif Q(F/Y)GF, conserved in other non-LTR retrotransposons, but not in retrovirus or LTR retrotransposon RTs. The remarkable conservation of this distinctive domain of non-LTR retrotransposon RTs suggests it has a vital and possibly unique role in the mode of reverse transcription of this class of transposon.

Amino Acid Sequence↗
PubMed1992

A kinetoplast DNA probe diagnostic for Leishmania infantum.

A 196 bp fragment of cloned kinetoplast (k) DNA of Leishmania infantum has been sequenced and shown to be diagnostic for this species. The DNA from 10(4)-10(5) promastigotes was routinely and specifically detected in crude dot blots of whole cells lysed in situ, and there was no cross-hybridization with 10(6) cells of L. major, L. tropica, L. killicki, L. aethiopica, L. braziliensis and L. mexicana. A low degree of hybridization was found with L. donovani and L. chagasi. This probe will be of particular use in Mediterranean countries, such as Italy, in which dermotropic zymodemes of L. infantum can be difficult to isolate and grow in numbers sufficient for isoenzyme typing.

Animals↗
PubMed1991

Retrotransposons and evolution in phlebotomines.

The polymerase chain reaction was used to amplify a segment of the reverse transcriptase (RT) gene of putative retrotransposons from Phlebotomus (Larroussius) perniciosus, P. (L.) perfiliewi, P. (Phlebotomus) papatasi and Lutzomyia (Lutzomyia) longipalpis. Based on amino acid sequence comparisons with known RT genes, the amplified products of these species were shown to be derived from non-LTR retrotransposons related to the F element of Drosophila melanogaster. The usefulness of this technique is discussed in relation to taxonomy and genetic manipulation.

Amino Acid Sequence↗
PubMed1990

Amplification and analysis of human DNA present in mosquito bloodmeals.

DNA fingerprinting should permit the identification of individual human hosts of haematophagous arthropods, providing epidemiologically useful information, for example, the biting rates on different people and the impact of insecticide-impregnated bednets. Investigations reported here demonstrate that it is possible to extract, amplify and fingerprint human DNA from the bloodmeals of individual female Anopheles gambiae mosquitoes kept at 24 degrees C for up to 10-15 h post-ingestion.

Animals↗
PubMed1989

A kinetoplast DNA probe diagnostic for Leishmania major: sequence homologies between regions of Leishmania minicircles.

A restriction fragment from a cloned kinetoplast minicircle DNA has been shown to be diagnostic for Leishmania major. This 402-bp TaqI fragment has been used routinely (as a radiolabelled probe) to detect 10(4) parasites in simple dot blots, both experimentally and in epidemiological surveys. It positively identified all stocks of L. major tested (including all six known zymodemes) and showed very low homology to kinetoplast DNA (kDNA) and chromosomal DNA of Leishmania infantum and Leishmania tropica, two species commonly isolated from patients and wild hosts within foci of L. major in the Old World. DNA sequence analysis of a minicircle of L. major is reported for the first time, and it is demonstrated that this species shares with Leishmania aethiopica, Sauroleishmania tarentolae and several species of Trypanosoma a region of conserved sequence that is involved in DNA replication, a process that could present targets for selective chemotherapeutic attack. Sequence and restriction fragment analyses have indicated the difficulties of selecting species-specific sequences from kDNA which, even in the same parasite clone, contains several predominant minicircle classes, not all of which contain diagnostic sequences.

Animals↗
PubMed1988

Natural infections of phlebotomine sandflies with Trypanosomatidae in central and south Italy.

In a survey of Leishmania infections of phlebotomine sandflies caught in central and south Italy in 1986, zymodeme MON-1 (Montpellier 1) of Leishmania infantum was isolated from Phlebotomus perniciosus and L. tarentolae [= Trypanosoma platydactyli?] was isolated from Sergentomyia minuta, providing for mainland Italy the first direct proof of these long-suspected parasite-vector associations. The roles of P. perfiliewi and P. perniciosus in the transmission of Leishmania spp. in Italy are discussed.

Animals↗
PubMed1988

DNA hybridizations on squash-blotted sandflies to identify both Phlebotomus papatasi and infecting Leishmania major.

Epidemiological field studies on leishmaniasis have been hampered by the laborious, and often inefficient, methods used to assess the rates of infection of sandfly vectors (Diptera; Phlebotominae) by species of the causative disease organisms, protozoal parasites of the genus Leishmania (Kinetoplastida; Trypanosomatidae). We report the rapid and accurate identification of both sandfly vector (Phlebotomus (Phlebotomus) papatasi (Scopoli] and infecting Leishmania major Yakimov & Schokov by DNA hybridizations to squash-blotted sandflies. Large numbers of whole (infected) sandflies can be quickly squashed on to nylon hybridization filters and (following standard procedures) the filter-bound DNA can be hybridized sequentially to cloned, multicopy genomic sequences that are specific for species of Leishmania (kinetoplast DNA) or for the sandfly (ribosomal (r) DNA). Our sandfly probe consists of a 3.2 kb fragment of the intergenic 'non-transcribed' spacer of rDNA of P. papatasi that we have detected only in this species: it is present in all six geographically isolated populations tested (from Tunisia through to India) but cannot be detected in the morphologically similar P. (Phlebotomus) duboscqi Neveu-Lemaire, the vector of Leishmania major south of the Sahara; it also cannot be detected in Phlebotomus species of the subgenera Larroussius and Paraphlebotomus that together with P. papatasi are the dominant man-biting sandflies in north African foci of zoonotic cutaneous leishmaniasis, where (as in many arid regions of western Asia) P. papatasi is believed to be the sole vector of L. major.

Animals↗
PubMed1983

Leishmaniasis in Brazil: XX. Prevalence of "enzootic rodent leishmaniasis" (Leishmania mexicana amazonensis), and apparent absence of "pian bois" (Le. braziliensis guyanensis), in plantations of introduced tree species and in other non-climax forests in eastern Amazônia.

In Amazonian Brazil most human leishmaniasis is due to Leishmania braziliensis s.l. and is acquired during the clearing of primary climax forest. One of the largest deforestation projects has taken place on the JARI property where plantations of exotic tree species are grown for paper pulp. The ability of the regional leishmaniasis enzootics to invade plantations was investigated. CDC light-trap catches indicated the phletobomine vectors of Le. b. guyanensis (causing "pian bois" in man) to be very scarce in JARI plantations compared to native-forest controls. It is concluded (drawing on other observations) that the vectors of "pian bois" are unlikely to thrive in any secondary forest. In contrast, catches from mammal traps and rodent-baited (Disney) traps demonstrated the presence in JARI plantations of infected Proechimys guyannensis and large populations of Lutzomyia flaviscutellata, respectively the major rodent reservoir and sandfly vector of Le. mexicana amazonensis. Alone amongst the local vectors of human cutaneous leishmaniasis, Lu. flaviscutellata is adapted to non-climax forests (primary or secondary, natural or man-made; synopsis given). It is predicted that the public health importance of Le. m. amazonensis is unlikely to diminish following the development of Amazônia. This is worrying because ca. 30% of Le. m. amazonensis infections in man cause highly-disfiguring, incurable "diffuse cutaneous leishmaniasis".

Animals↗
PubMed1981

Leishmaniasis in Brazil: XVI. Isolation and identification of Leishmania species from sandflies, wild mammals and man in north Para State, with particular reference to L. braziliensis guyanensis causative agent of "pian-bois".

A total of 125 wild mammals (14 different species) were examined for evidence of infection with Leishmania in an area of primary forest highly endemic for "pian-bois", due to Leishmania braziliensis guyanensis, in north Pará State, Brazil. Parasites isolated were characterized biologically, and biochemically on enzymic profiles. L. b. guyanensis was isolated from the viscera of one lesser anteater (Tamandua tetradactyla) and one opossum (Didelphis marsupialis), and the skin of one rodent (Proechimys guyannensis). The isolates were indistinguishable from 10 others previously made from the sandfly vectors Lutzomyia umbratilis (five) and Lu. whitmani (five), and nine isolates from field-workers who became infected during these studies. Leishmania mexicana amazonensis was obtained from the skin of 21 animals, including three species of opossums (D. marsupialis, Philander opossum and Metachirus nudicaudatus) and two species of rodents (proechimys guyannensis and Dasyprocta sp.). A peripylarian Leishmania isolated from the viscera of two armadillos (Dasypus novemcinctus) was shown to be different, biologically and biochemically, from L. b. guyanensis and L. m. amazonensis. Four other isolates of Leishmania, from the rodents Rhipidomys leucodactylus (one) and P. guyannensis (three) have yet to be characterized owing to their very poor growth in both hamster skin and in vitro culture: they appear closest, however, to L. braziliensis braziliensis. The complexity of Amazonian leishmaniasis is discussed, and attention drawn to the importance of edentates as reservoir hosts of some leishmanias in the New World. Whereas L. mexicana subspecies appear largely restricted to the skin of their natural hosts, subspecies of L. braziliensis are commonly found in the viscera.

Animal Population Groups↗