[leish-l] Fwd: Articles found by RefScout 2005/08/24 - 2005/34

[jeffreyj at usp.br]

   Date: Wed, 24 Aug 2005 03:34:46
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REQUEST: [ leishmaniasis ]

(13 articles match this request)



PMID: 16095641
 

TITLE: Usefulness of PCR in the diagnosis of cutaneous leishmaniasis in
Tunisia.

AUTHORS: N Chargui, P Bastien, K Kallel, N Haouas, F Messaidi Akrout, A
Masmoudi, J Zili, E Chaker, A Dhahri Ben Othman, R Azaiez, L Crobu, H Mezhoud,
H Babba

AFFILIATION: Laboratoire de Parasitologie-Mycologie, 99-UR/08-05, Faculté de
Pharmacie de Monastir, Rue Avicenne, Monastir 5000, Tunisia.

REFERENCE: Trans R Soc Trop Med Hyg 2005 Oct 99(10):762-8

We assessed the efficiency of a PCR method in establishing the diagnosis
 of cutaneous leishmaniasis (CL) in Tunisian patients. Four hundred and 
thirty specimens collected passively from patients with cutaneous ulcers
 suggestive of leishmaniasis attending health centres for diagnosis were
 included in the study. Dermal scrapings were analysed both by 
parasitological (examination of Giemsa-stained smears and in vitro 
cultivation) methods and by a genus-specific PCR detecting a fragment of
 the 18S rRNA gene. Microscopy revealed amastigotes in 245 samples (57.0
%) and in vitro cultivation gave positive results in 88 cases (20.5%), 
whereas PCR detected Leishmania in 301 samples (70%). The sensitivities 
inferred from our results were 99.3%, 80.8% and 29% for PCR, microscopic
 examination and in vitro cultivation, respectively. The different forms
 of CL in this country are caused by three species of Leishmania and are
 treated with the same protocol. Of 303 well-documented cases in our 
study, 99% were probably caused by Leishmania major and 1% by Leishmania
 infantum. The lack of species-specific diagnosis is not known to affect
 treatment or prognosis in Tunisia. These data support the incorporation
 of PCR into diagnostic strategies for CL, particularly in Tunisia.








PMID: 16113329
 

TITLE: Arginase I Induction during Leishmania major Infection Mediates the
Development of Disease.

AUTHORS: Virginia Iniesta, Jesualdo Carcelén, Isabel Molano, Pablo M V
Peixoto,
Eloy Redondo, Pilar Parra, Marina Mangas, Isabel Monroy, Maria Luisa Campo,
Carlos Gómez Nieto, Inés Corraliza

AFFILIATION: Facultad de Veterinaria, Universidad de Extremadura. Avda. de la
Universidad s/n, Cáceres 10071, Spain. corragen at unex.es.

REFERENCE: Infect Immun 2005 Sep 73(9):6085-90

In a previous work, we demonstrated that the induction of arginase I 
favored the replication of Leishmania inside macrophages. Now we have 
analyzed the differential expression of this enzyme in the mouse model 
of L. major infection. Ours results show that arginase I is induced in 
both susceptible and resistant mice during the development of the 
disease. However, in BALB/c-infected tissues, the induction of this 
protein parallels the time of infection, while in C57BL/6 mice, the 
enzyme is upregulated only during footpad swelling. The induction of the
 host arginase in both strains is mediated by the balance between 
interleukin-4 (IL-4) and IL-12 and opposite to nitric oxide synthase II 
expression. Moreover, inhibition of arginase reduces the number of 
parasites and delays disease outcome in BALB/c mice, while treatment 
with l-ornithine increases the susceptibility of C57BL/6 mice. Therefore
, arginase I induction could be considered a marker of disease in 
leishmaniasis.




PMID: 16113303
 

TITLE: Vaccination with the Leishmania infantum Acidic Ribosomal P0 Protein
plus
CpG Oligodeoxynucleotides Induces Protection against Cutaneous Leishmaniasis in
C57BL/6 Mice but Does Not Prevent Progressive Disease in BALB/c Mice.

AUTHORS: Salvador Iborra, Javier Carrión, Charles Anderson, Carlos Alonso,
David Sacks, Manuel Soto

AFFILIATION: Centro de Biología Molecular "Severo Ochoa," Universidad
Autónoma
de Madrid, 28049 Madrid, Spain. msoto at cbm.uam.es.

REFERENCE: Infect Immun 2005 Sep 73(9):5842-52

We have examined the efficacy of the administration in mice of a 
molecularly defined vaccine based on the Leishmania infantum acidic 
ribosomal protein P0 (rLiP0). Two different challenge models of murine 
cutaneous leishmaniasis were used: (i) subcutaneous inoculation of L. 
major parasites in susceptible BALB/c mice (a model widely used for 
vaccination analysis) and (ii) the intradermal inoculation of a low 
infective dose in resistant C57BL/6 mice (a model that more accurately 
reproduces the L. major infection in natural reservoirs and in human 
hosts). First, we demonstrated that C57BL/6 mice vaccinated with LiP0-
DNA or rLiP0 protein plus CpG oligodeoxynucleotides (ODN) were protected
 against the development of dermal pathology and showed a reduction in 
the parasite load. This protection was associated with production of 
gamma interferon (IFN-gamma) in the dermal site. Secondly, we showed 
that immunization with rLiP0 plus CpG ODN is able to induce only partial
 protection in BALB/c, since these mice finally developed a progressive 
disease. Further, we demonstrated that LiP0 vaccination induces a Th1 
immunological response in both strains of mice. In both cases, the 
antibodies against LiP0 were predominantly of the immunoglobulin G2a 
isotype, which was correlated with an rLiP0-stimulated production of IFN
-gamma in draining lymph nodes. Finally, we demonstrated that LiP0 
vaccination does not prevent the Th2 response induced by L. major 
infection in BALB/c mice. Taken together, these data indicate that the 
BALB/c model of cutaneous leishmaniasis may undervalue the potential 
efficacy of some vaccines based on defined proteins, making C57BL/6 a 
suitable alternative model to test vaccine candidates.




PMID: 16113301
 

TITLE: Toward a Novel Experimental Model of Infection To Study American
Cutaneous Leishmaniasis Caused by Leishmania braziliensis.

AUTHORS: Tatiana R de Moura, Fernanda O Novais, Fabiano Oliveira, Jorge
Clarêncio, Almério Noronha, Aldina Barral, Claudia Brodskyn, Camila I de
Oliveira

AFFILIATION: Centro de Pesquisas Gonçalo Moniz, Fundação Oswaldo Cruz, Rua
Waldemar Falcão, 121, Salvador, BA 40295-001, Brazil.
camila at cpqgm.fiocruz.br.

REFERENCE: Infect Immun 2005 Sep 73(9):5827-34

Leishmania spp. cause a broad spectrum of diseases collectively known as
 leishmaniasis. Leishmania braziliensis is the main etiological agent of
 American cutaneous leishmaniasis (ACL) and mucocutaneous leishmaniasis
. In the present study, we have developed an experimental model of 
infection that closely resembles ACL caused by L. braziliensis. In order
 to do so, BALB/c mice were infected in the ear dermis with 10(5) 
parasites and distinct aspects of the infection were evaluated. 
Following inoculation, parasite expansion in the ear dermis was 
accompanied by the development of an ulcerated dermal lesion which 
healed spontaneously, as seen by the presence of a scar. Histological 
analysis of infected ears showed the presence of a mixed inflammatory 
infiltrate consisting of both mononuclear and polymorphonuclear cells. 
In draining lymph nodes, parasite replication was detected throughout 
the infection. In vitro restimulation of draining lymph node cells 
followed by intracellular staining showed an up-regulation in the 
production of gamma interferon (IFN-gamma) and in the frequency of IFN-
gamma-secreting CD4(+) and CD8(+) T cells. Reverse transcription-PCR of 
ears and draining lymph node cells showed the expression of CC 
chemokines. The dermal model of infection with L. braziliensis herein is
 able to reproduce aspects of the natural infection, such as the 
presence of an ulcerated lesion, parasite dissemination to lymphoid 
areas, and the development of a Th1-type immune response. These results 
indicate that this model shall be useful to address questions related to
 the concomitant immunity to reinfection and parasite persistence 
leading to mucocutaneous leishmaniasis.




PMID: 16103587
 

TITLE: Detection of leishmanial antigen in the urine of patients with visceral
leishmaniasis by a latex agglutination test.

AUTHORS: Shyam Sundar, Shrinkhla Agrawal, Kalpana Pai, Michael Chance, Marcel
Hommel

AFFILIATION: Kala-Azar Medical Research Center, Department of Medicine,
Institute of Medical Sciences, Banaras Hindu University, Varanasi, India;
Liverpool School of Tropical Medicine, Liverpool, United Kingdom.

REFERENCE: Am J Trop Med Hyg 2005 Aug 73(2):269-71

Diagnosis of visceral leishmaniasis (VL) is usually done by 
demonstration of parasites in tissue smears. However, obtaining these 
smears may be risky, painful, and difficult. Antibody-based diagnostics 
are limited by their inability to predict active disease. In this study
, a new latex agglutination test (KAtex), which detects parasite antigen
 in freshly voided and boiled urine, was evaluated in patients with VL 
before the start (n = 382) and at the end of treatment (n = 273); 185 
healthy controls from leishmaniasis-endemic region were also studied. 
The KAtex result was positive in 87% (95% confidence interval [CI] = 83.

3-90.3). However, at the end of treatment only 3% (95% CI = 1.6-6.2) 
patients were positive. The specificity of the test was 99% and 2 of 185
 healthy controls tested positive. Positive and negative predictive 
values were 0.994 and 0.788, respectively. KAtex is a promising test, 
and in a simplified and improved format it could be applied meaningfully
 in the diagnosis of VL.




PMID: 16101793
 

TITLE: Treatment of canine Old World visceral leishmaniasis: a systematic
review.

AUTHORS: Chiara Noli, Silvia T Auxilia

AFFILIATION: Ospedale Veterinario Cuneese, Via Cuneo 52/N, 12011 Borgo S.
Dalmazzo (CN), Italy.

REFERENCE: Vet Dermatol 2005 Aug 16(4):213-32

Canine visceral leishmaniasis is a systemic disease caused by Leishmania
 infantum. The aim of this systematic review was to identify and 
evaluate the evidence of efficacy of interventions for treatment or 
prevention of canine visceral leishmaniasis, and to propose 
recommendations for or against their use. Forty-seven articles 
describing clinical trials published between 1980 and 2004 fulfilled 
selection criteria. The evaluation of clinical trials provided good 
evidence for recommending the use of meglumine antimoniate at a minimum 
dosage of 100 mg kg(-1) daily for at least 3-4 weeks, combined with 
allopurinol in order to obtain a good clinical efficacy and a reduced 
relapse rate. The evaluation of the articles also provided fair evidence
 for recommending the use of pentamidine (4 mg kg(-1) twice weekly) and 
aminosidine (5 mg kg(-1) twice daily) for 3-4 weeks. There was 
insufficient evidence for recommending the use of allopurinol alone, 
amphotericin B, buparvaquone, ketoconazole, enrofloxacin, and the 
combinations of metronidazole with spiramicyn or metronidazole with 
enrofloxacin. Fair evidence against the use of aminosidine at high 
dosages (20-80 mg kg(-1) per day) was proposed due to its side effects. 
Evaluation of articles on repellent measures against sand fly vectors of
 leishmaniasis provided good evidence for recommending deltamethrin 
collars and fair evidence for recommending spot-on permethrin.








PMID: 16113906
 

TITLE: [Phlebotomine sandflies of Southern Brazil.]

AUTHORS: Rubens Massafera, Allan Martins da Silva, Antônio Plácido de
Carvalho, Demilson Rodrigues Dos Santos, Eunice Aparecida Bianchi Galati,
Ueslei Teodoro

AFFILIATION: Núcleo de Entomologia de Jacarezinho, Secretaria de Estado da
Saúde do Paraná, Jacarezinho, PR, Brasil.

REFERENCE: Rev Saude Publica 2005 Aug 39(4):571-7

OBJECTIVE: To identify the sandfly fauna and some aspects of their 
behavior in forest and anthropic environments. METHODS: Sandfly captures
 were undertaken in farm (23 masculine 6' S; 50 masculine 22' W), in 
Southern Brazil. Falcão light traps were set in forest, domicile and 
domestic animal shelters and mosquitoes were collected monthly, between 
17h and 7h, from March 1997 to February 1998. RESULTS: A total of 3,655 
specimens representing 13 species were captured. Nyssomyia whitmani and 
Nyssomyia neivai were the predominant species, with a total of 2,977 
specimens (81.0%). Of these two species, a total of 2,552 (85.7%) 
specimens were captured in intradomiciliary and peridomiciliary 
environments, 2,332 (91.3%) of them in a pigsty. These two species 
predominated between 20h-21h when 90.4% of the specimens were captured. 
CONCLUSIONS: Five sandfly species, N. whitmani, N. neivai, Migonemyia 
migonei, Pintomyia pessoai and Pintomyia fischeri, potential vectors of 
cutaneous leishmaniasis, were captured. The importance of the two former
 species is emphasized, since both presented similar behavior in regard 
to seasonal period, hourly frequency and predominance in the anthropic 
environment. Besides, N. whitmani was the most predominant species.




PMID: 16103589
 

TITLE: The value of a new microculture method for diagnosis of visceral
leishmaniasis by using bone marrow and peripheral blood.

AUTHORS: Adil M Allahverdiyev, Malahat Bagirova, Soner Uzun, Derya Alabaz,
Necmi
Aksaray, Emine Kocabas, Fatih Koksal

AFFILIATION: Cukurova University, Tropical Diseases Research Centre, Adana,
Turkey; Cukurova University, School of Medicine, Department of Pediatrics,
Infectious Diseases Unit, Adana, Turkey.

REFERENCE: Am J Trop Med Hyg 2005 Aug 73(2):276-80

We have demonstrated that the microculture method (MCM) enables the 
diagnosis of visceral leishmaniasis (VL) with samples from both the bone
 marrow (BM) and peripheral blood (PB). The MCM is superior to the 
traditional culture method (TCM) as determined by its higher sensitivity
 in the detection of promastigotes and the more rapid time for emergence
 of promastigotes. The sensitivity of MCM (100% in BMs and 77.8-100% in 
PB) was considerably higher than that of the TCM (37.5-100% in BMs and 0
-100% in PB) according to decreasing parasite density (P < 0.05). The
 concentration of parasites in buffy coats has increased the sensitivity
 of both methods, especially that of the MCM. Detection of promastigotes
 by MCM requires lower amounts of culture media (25-50 muL) and shorter 
incubation periods (2-7 days) than TCM (2.5-3.5 mL and 15-35 days, 
respectively). MCM was found to be valuable with the advantages of 
simplicity and sensitivity, in addition to being cost-effective in the 
routine diagnosis for VL in Adana Turkey.




PMID: 16098272
 

TITLE: Leishmaniasis cutis: report of two cases.

AUTHORS: Ke-jian Xu, Yue-hua Liu, Kai Fang

AFFILIATION: Department of Dermatology, Peking Union Medical College Hospital,
Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing
100730, China.

REFERENCE: Chin Med J (Engl) 2005 Jul 118(13):1137-9




PMID: 16105358
 

TITLE: A new focus for cutaneous leishmaniasis in the West Coast of Turkey.

AUTHORS: Hatice Ertabaklar, S Oncü, S Ertug

AFFILIATION: Department of Parasitology, Adnan Menderes University Medical
Faculty, Aydin, Turkey.

REFERENCE: Trop Doct 2005 Jul 35(3):189




PMID: 15946800
 

TITLE: A seroepidemiologic survey of canine visceral leishmaniosis among
apparently healthy dogs in Croatia.

AUTHORS: T Zivicnjak, F Martinković, A Marinculić, V Mrljak, N Kucer, V
Matijatko, Z Mihaljević, R Barić-Rafaj

AFFILIATION: Department for Parasitology and Parasitic Diseases, Veterinary
Faculty, University of Zagreb, Heinzelova 55, 10000 Zagreb, Croatia.
tatjanaz at vef.hr

REFERENCE: Vet Parasitol 2005 Jul 131(1-2):35-43

Cross-sectional investigation was done on seroprevalence of Leishmania 
sp. infection among apparently healthy dogs in an area where canine 
leishmaniosis is endemic. Survey included 68 dogs living in the coastal 
city of Split, and 238 dogs living in 12 villages scattered in the 
hinterland. Each dog was clinically examined for the presence of some 
discrete signs compatible with leishmaniosis and by dot-ELISA 
modification determined the presence of anti-Leishmania antibodies. The 
titre 1:600 and higher was regarded as positive in the study. The 
seroprevalence ranged from 0 to 42.85%, depending on the location. 54.34
% of the seropositive dogs had moderately enlarged lymph nodes and/or 
some discrete changes on the skin. In our parasitological study, 
Leishmania sp. was isolated from several seropositive animals that had 
some clinical signs and from a few which did not have any. Data analysis
 revealed that serological positivity to Leishmania sp. was not 
associated with a dog's outdoor lifestyle and utility, but was 
associated with the gender and age.








PMID: 16108545
 

TITLE: Axenic promastigote forms of Leishmania (Viannia) lainsoni as an
alternative source for Leishmania antigen production.

AUTHORS: José R Corrêa, Sidney G Santos, Márcio S Araújo, Cibele Baptista,
Maurilio J Soares, Reginaldo P Brazil

AFFILIATION: Lab. Biologia Celular de Microrganismos, Departamento de
Ultra-estrutura e Biologia Celular, Instituto Oswaldo Cruz/FIOCRUZ, Av. Brasil
4365, 21040-900 Rio de Janeiro, RJ, Brazil. correa at ioc.fiocruz.br

REFERENCE: J Parasitol 2005 Jun 91(3):551-6

The present study demonstrates that axenic cultures of Leishmania (
Viannia) lainsoni produce larger cell masses in NNN-LIT medium, as well 
as higher amounts of total proteins in cell extracts, than Leishmania (
Leishmania) amazonensis. Antigenicity of L. (V.) lainsoni whole 
promastigotes is similar to that of L. (L.) amazonensis, as demonstrated
 by an indirect immunofluorescence diagnostic test using sera from human
 patients and dogs infected with visceral leishmaniasis. Infectivity of 
the L. (V.) lainsoni strain used in the present work was demonstrated by
 the detection by transmission-electron microscopy of tissue amastigotes
 in skin lesion samples from an experimentally infected hamster. 
Incubation of lesion fragments in NNN-LIT medium allowed us to obtain 
promastigote forms, which could be cultivated successfully in vitro. 
lsoenzyme analysis of such promastigotes confirmed the parasite strain 
as L. (V.) lainsoni, as compared to other Leishmania reference strains. 
Our data indicate that L. (V.) lainsoni is a useful alternative source 
for antigen production as well for use in assays that depend on large 
cell volumes of Leishmania spp. parasites.




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PMID: 15855481
 

TITLE: Antileishmanial activity of the terpene nerolidol.

AUTHORS: Denise C Arruda, Fabio Luiz D'Alexandri, Alejandro M Katzin, Silvia R
B
Uliana

AFFILIATION: Departamento de Parasitologia, Instituto de Ciências Biomédicas,
Universidade de São Paulo, Av. Professor Lineu Prestes, 1374, CEP 05508-900,
São Paulo, SP, Brazil.

REFERENCE: Antimicrob Agents Chemother 2005 May 49(5):1679-87

The activity of nerolidol, a sesquiterpene used as a food-flavoring 
agent and currently under testing as a skin penetration enhancer for the
 transdermal delivery of therapeutic drugs, was evaluated against 
Leishmania species. Nerolidol inhibited the growth of Leishmania 
amazonensis, L. braziliensis, and L. chagasi promastigotes and L. 
amazonensis amastigotes with in vitro 50% inhibitory concentrations of 
85, 74, 75, and 67 microM, respectively. The treatment of L. amazonensis
-infected macrophages with 100 microM nerolidol resulted in 95% 
reduction in infection rates. Inhibition of isoprenoid biosynthesis, as 
shown by reduced incorporation of [2-(14)C]mevalonic acid (MVA) or [1-(
14)C]acetic acid precursors into dolichol, ergosterol, and ubiquinone, 
was observed in nerolidol-treated promastigotes. This drug effect can be
 attributed to the blockage of an early step in the mevalonate pathway, 
since incorporation of the precursor [1(n)-(3)H]farnesyl pyrophosphate 
in polyisoprenoids is not inhibited by nerolidol. L. amazonensis-
infected BALB/c mice were treated with intraperitoneal doses of 100 mg/
kg/day for 12 days or topically with 5 or 10% ointments for 4 weeks. 
Significant reduction of lesion sizes in nerolidol treated mice was 
observed for both treatment routes. However, long-term follow up 
indicated that the disease was not cured in this highly susceptible 
animal model. Nonetheless, the in vitro activity of nerolidol against 
these parasites may prove a useful tool for the development of new drugs
 for the treatment of leishmaniasis. In addition, biosynthesis of 
dolichols with 11 and 12 isoprene units was identified in Leishmania, as
 described for other trypanosomatids and Apicomplexa.




REQUEST: [ leishmania ]

(16 articles match this request. 7 articles matching other requests removed)



PMID: 16081295
 

TITLE: Synthesis and antileishmanial activities of 4,5-di-substituted acridines
as compared to their 4-mono-substituted homologues.

AUTHORS: Di Giorgio Carole, De Méo Michel, Chiron Julien, Delmas Florence,
Nikoyan Anna, Jean Séverine, Dumenil Gérard, Timon-David Pierre, Galy
Jean-Pierre

AFFILIATION: Laboratoire de Parasitologie, Hygiène et Zoologie, Faculté de
Pharmacie, 27 Bd. Jean Moulin, 13385 Marseille Cedex 05, France.

REFERENCE: Bioorg Med Chem 2005 Oct 13(19):5560-8

Newly synthesized 4,5-di-substituted acridines were assessed for in 
vitro antileishmanial activities as compared to those of their 4-mono-
substituted homologues. Mono-substituted acridines exhibited a weak 
specificity for Leishmania parasites. Di-substituted acridines, on the 
contrary, displayed interesting amastigote-specific activities through a
 mechanism of action that might not involve intercalation to DNA. This 
antileishmanial property, associated with a low antiproliferative 
activity towards human cells, led to the identification of a new class 
of promising acridine derivatives such as 4,5-bis(hydroxymethyl)acridine
 with a nonclassical mechanism of action based on the inhibition of 
Leishmania internalization within macrophages. In the meantime, the 
effects of experimental lighting on the biological properties of 
acridines were assessed: experimental lighting did not significantly 
improve the antileishmanial activity of the compounds since it produced 
a greater toxicity against human cells.




PMID: 15850459
 

TITLE: Leishmania major encodes an unusual peroxidase that is a close homologue
of plant ascorbate peroxidase: a novel role of the transmembrane domain.

AUTHORS: Subrata Adak, Alok K Datta

AFFILIATION: Division of Infectious Diseases, Leishmania Group, Indian
Institute
of Chemical Biology, 4, Raja S. C. Mullick Road, Kolkata 700 032, India.

REFERENCE: Biochem J 2005 Sep 390(Pt 2):465-74

Haem-containing enzymes (peroxidase and catalase) are widely distributed
 among prokaryotes and eukaryotes and play a vital role in H(2)O(2) 
detoxification. But, to date, no haem-containing enzymatic defence 
against toxic H(2)O(2) has been discovered in Leishmania species. We 
cloned, expressed and purified an unusual plant-like APX (ascorbate 
peroxidase) from Leishmania major (LmAPX) and characterized its 
catalytic parameters under steady-state conditions. Examination of its 
protein sequence indicated approx. 30-60% identity with other APXs. The 
N-terminal extension of LmAPX is characterized by a charged region 
followed by a stretch of 22 amino acids containing a transmembrane 
domain. To understand how the transmembrane domain influences the 
structure-function of LmAPX, we generated, purified and extensively 
characterized a variant that lacked the transmembrane domain. 
Eliminating the transmembrane domain had no impact on substrate-binding 
affinity but slowed down ascorbate oxidation and increased resistance to
 H(2)O(2)-dependent inactivation in the absence of electron donor by 480
-fold. Spectral studies show that H(2)O(2) can quickly oxidize the 
native enzyme to compound (II), which subsequently is reduced back to 
the native enzyme by an electron donor. In contrast, ascorbate-free 
transmembrane domain-containing enzyme did not react with H(2)O(2), as 
revealed by the absence of compound (II) formation. Our findings suggest
 that the single copy LmAPX gene may play an important role in 
detoxification of H(2)O(2) that is generated by endogenous processes and
 as a result of external influences such as the oxidative burst of 
infected host macrophages or during drug metabolism by Leishmania.




PMID: 16113310
 

TITLE: Transforming Growth Factor {beta}1 Production by CD4+ CD25+ Regulatory T
Cells in Peripheral Blood Mononuclear Cells from Healthy Subjects Stimulated
with Leishmania guyanensis.

AUTHORS: A Kariminia, E Bourreau, H Pascalis, P Couppié, D Sainte-Marie, F
Tacchini-Cottier, P Launois

AFFILIATION: WHO-IRTC, Department of Biochemistry, University of Lausanne, Ch
des Boveresses, 155, 1066 Epalinges, Switzerland. pascal.launois at unil.ib.ch.

REFERENCE: Infect Immun 2005 Sep 73(9):5908-14

Transforming growth factor beta (TGF-beta) has been shown to be a 
central immunomodulator used by leishmaniae to escape effective 
mechanisms of protection in human and murine infections with these 
parasites. However, all the information is derived from studies of 
established infection, while little is known about TGF-beta production 
in response to Leishmania stimulation in healthy subjects. In this study
, TGF-beta1 production was demonstrated in peripheral blood mononuclear 
cells from healthy subjects never exposed to leishmaniae in response to 
live Leishmania guyanensis, and the TGF-beta1-producing cells were 
described as a distinct subpopulation of CD4(+) CD25(+) regulatory T 
cells. The suppressive properties of CD4(+) CD25(+) T cells were 
demonstrated in vitro by their inhibition of production of interleukin 2
 (IL-2) and IL-10 by CD4(+) CD25(-) T cells in the presence of either 
anti-CD3 or L. guyanensis. Although neutralization of TGF-beta1 did not 
reverse the suppressive activity of CD4(+) CD25(+) T cells activated by 
anti-CD3, it reversed the suppressive activity of CD4(+) CD25(+) T cells
 activated by L. guyanensis. Altogether our data demonstrated that TGF-
beta1 is involved in the suppressive activity of L. guyanensis-
stimulated CD4(+) CD25(+) T cells from healthy controls.








PMID: 16107969
 

TITLE: Turnover of Neutrophils Mediated by Fas Ligand Drives Leishmania major
Infection.

AUTHORS: Flavia L Ribeiro-Gomes, Maria Carolina A Moniz-de-Souza, Valeria M
Borges, Marise P Nunes, Marcio Mantuano-Barradas, Heloisa D'Avila, Patricia T
Bozza, Vera L Calich, George A Dosreis

AFFILIATION: Instituto de Biofisica Carlos Chagas Filho, Federal University of
Rio de Janeiro, Brazil.

REFERENCE: J Infect Dis 2005 Sep 192(6):1127-34

Apoptosis mediated by Fas ligand (FasL) initiates inflammation 
characterized by neutrophilic infiltration. Neutrophils undergo 
apoptosis and are ingested by macrophages. Clearance of dead neutrophils
 leads to prostaglandin- and transforming growth factor- beta -dependent
 replication of Leishmania major in macrophages from susceptible mice. 
How L. major induces neutrophil turnover in a physiological setting is 
unknown. We show that BALB/c FasL-sufficient mice are more susceptible 
to L. major infection than are FasL-deficient mice. FasL promotes the 
apoptosis of infected resident macrophages and attracts neutrophils. 
Furthermore, FasL-sufficient neutrophils exacerbate L. major replication
 in macrophages, whereas FasL-deficient neutrophils induce parasite 
killing. These contrasting effects are due to delaying apoptosis and the
 clearance of FasL-deficient neutrophils. The transfer of neutrophils 
exacerbates infection in FasL-sufficient mice but reduces infection in 
FasL-deficient mice. Depletion of neutrophils abolishes the 
susceptibility of FasL-sufficient mice. These data illustrate a 
deleterious role of the FasL-mediated turnover of neutrophils on L. 
major infection.




PMID: 15901238
 

TITLE: Characterization of the ATPase activity of topoisomerase II from
Leishmania donovani and identification of residues conferring resistance to
etoposide.

AUTHORS: Tanushri Sengupta, Mandira Mukherjee, Aditi Das, Chhabinath Mandal,
Rakhee Das, Tanmoy Mukherjee, Hemanta K Majumder

AFFILIATION: Molecular Parasitology Laboratory, Indian Institute of Chemical
Biology, Kolkata-700032, India.

REFERENCE: Biochem J 2005 Sep 390(Pt 2):419-26

We have cloned and expressed the 43 kDa N-terminal domain of Leishmania 
donovani topoisomerase II. This protein has an intrinsic ATPase activity
 and obeys Michaelis-Menten kinetics. Cross-linking studies indicate 
that the N-terminal domain exists as a dimer both in the presence and 
absence of nucleotides. Etoposide, an effective antitumour drug, traps 
eukaryotic DNA topoisomerase II in a covalent complex with DNA. In the 
present study, we report for the first time that etoposide inhibits the 
ATPase activity of the recombinant N-terminal domain of L. donovani 
topoisomerase II. We have modelled the structure of this 43 kDa protein 
and performed molecular docking analysis with the drug. Mutagenesis of 
critical amino acids in the vicinity of the ligand-binding pocket 
reveals less efficient inhibition of the ATPase activity of the enzyme 
by etoposide. Taken together, these results provide an insight for the 
development of newer therapeutic agents with specific selectivity.




PMID: 16101794
 

TITLE: PCR technique detection of Leishmania spp. but not Mycobacterium spp. in
canine cutaneous 'sterile' pyogranuloma/granuloma syndrome.

AUTHORS: L Cornegliani, D Fondevila, A Vercelli, G Mantero, A Fondati

AFFILIATION: Ambulatorio Veterinario Associato, Corso Traiano 99/d, Torino,
Italy.

REFERENCE: Vet Dermatol 2005 Aug 16(4):233-8

Cutaneous 'sterile' pyogranuloma/granuloma syndrome (SPGS) is an 
uncommon canine skin disorder of unknown aetiopathogenesis. 
Histopathological findings and failure to demonstrate an aetiologic 
agent are suggestive of this syndrome. Nevertheless, it has been 
hypothesized that SPGS may be related to an immune response against 
persistent endogenous or exogenous antigens. The presence of Leishmania 
and Mycobacterium organisms was investigated by polymerase chain 
reaction (PCR) techniques in 46 canine skin samples histopathologically 
diagnosed as SPGS. Concomitantly, an immunohistochemical technique for 
Leishmania detection was applied on the same samples and the results 
were compared with those from PCR. The PCR technique yielded positive 
results for Leishmania spp. in 21 out of 46 skin samples. The results of
 immunohistochemical techniques were identical to those obtained by PCR
. The PCR technique gave negative results for Mycobacterium spp. in all 
the samples examined. These results suggest the importance of looking 
for Leishmania spp. in skin biopsies with histopathological findings 
consistent with the diagnosis of SPGS.




PMID: 15955815
 

TITLE: Down-regulation of 7SL RNA expression and impairment of vesicular
protein
transport pathways by Leishmania infection of macrophages.

AUTHORS: Smita Misra, Manish K Tripathi, Gautam Chaudhuri

AFFILIATION: Division of Microbial Pathogenesis and Immune Response, Department
of Biomedical Sciences, Meharry Medical College, Nashville, TN 37208, USA.

REFERENCE: J Biol Chem 2005 Aug 280(32):29364-73

The parasitic protozoan Leishmania specifically manipulates the 
expression of host macrophage genes during initial interactions, as 
revealed by mRNA differential display reverse transcription-PCR and cDNA
 microarray analyses. The genes that are down-regulated in mouse (J774G8
) or human (U937) macrophages upon exposure to Leishmania include small 
RNA transcripts from the short interspersed element sequences. Among the
 short interspersed element RNAs that are down-regulated is 7SL RNA, 
which is the RNA component of the signal recognition particle. Because 
the microbicidal functions of macrophages profoundly count on vesicular 
protein transport processes, down-regulation of 7SL RNA may be 
significant in the establishment of infection by Leishmania in 
macrophage phagolysosomes. To evaluate whether down-regulation of 7SL 
RNA results in inhibition of signal recognition particle-mediated 
vesicular protein transport processes, we have tested and found that the
 targeting of proteins to the endoplasmic reticulum and plasma membrane 
and the secretion of proteins by macrophages are compromised in 
Leishmania-infected J774G8 and U937 cells. Knocking down 7SL RNA using 
small interfering RNA mimicked the effect of exposure of macrophages to 
Leishmania. The overexpression of 7SL RNA in J774G8 or U937 cells made 
these cells resistant to Leishmania infection, suggesting the possible 
biological significance of down-regulation of 7SL RNA synthesis in the 
establishment of infection by Leishmania. We conclude that Leishmania 
down-regulates 7SL RNA in macrophages to manipulate the targeting of 
many proteins that use the vesicular transport pathway and thus favors 
its successful establishment of infection in macrophages.




PMID: 16113885
 

TITLE: Subcellular localization of an intracellular serine protease of 68 kDa
in
Leishmania (Leishmania) amazonensis promastigotes.

AUTHORS: José Andrés Morgado-Díaz, Raquel Elisa da Silva-Lopez, Carlos
Roberto Alves, Maurilio José Soares, Suzana Corte-Real, Salvatore Giovanni De
Simone

AFFILIATION: Grupo de Biologia Estrutural, Divisão de Biologia Celular, Centro
de Pesquisas, Instituto Nacional de Câncer, Rio de Janeiro, RJ, Brasil.

REFERENCE: Mem Inst Oswaldo Cruz 2005 Jul 100(4):377-83

Here we report the subcellular localization of an intracellular serine 
protease of 68 kDa in axenic promastigotes of Leishmania (Leishmania) 
amazonensis, using subcellular fractionation, enzymatic assays, 
immunoblotting, and immunocytochemistry. All fractions were evaluated by
 transmission electron microscopy and the serine protease activity was 
measured during the cell fractionation procedure using a-N-r-tosyl-L-
arginine methyl ester (L-TAME) as substrate, phenylmethylsulphone 
fluoride (PMSF) and L-1-tosylamino-2-phenylethylchloromethylketone (TPCK
) as specific inhibitors. The enzymatic activity was detected mainly in 
a membranous vesicular fraction (6.5-fold enrichment relative to the 
whole homogenate), but also in a crude plasma membrane fraction (2.0-
fold). Analysis by SDS-PAGE gelatin under reducing conditions 
demonstrated that the major proteolytic activity was found in a 68 kDa 
protein in all fractions studied. A protein with identical molecular 
weight was also recognized in immunoblots by a polyclonal antibody 
against serine protease (anti-SP), with higher immunoreactivity in the 
vesicular fraction. Electron microscopic immunolocalization using the 
same polyclonal antibody showed the enzyme present at the cell surface, 
as well as in cytoplasmic membranous compartments of the parasite. Our 
findings indicate that the internal location of this serine protease in 
L. amazonensis is mainly restricted to the membranes of intracellular 
compartments resembling endocytic/exocytic elements.




PMID: 16113874
 

TITLE: Leishmania (Viannia) braziliensis growth in vitro culture relies more on
folic acid availability than Leihsmania (Leishmania) amazonensis.

AUTHORS: Andrea Niño, Marcela Camacho

AFFILIATION: Laboratorio de Biofísica, Centro Internacional de Física,
Bogotá, Colombia.

REFERENCE: Mem Inst Oswaldo Cruz 2005 May 100(3):309-10

We compared the in vitro growth of promastigotes from two Leishmania 
species in TC-100 and Schneider media. Leishmania (Leishmania) 
amazonensis replication rates were similar in both tissue culture media 
and reached maximum rates by 48 h. In contrast Leishmania (Viannia) 
braziliensis growth was significantly greater in TC-100 but maximum 
rates were achieved by 96 h. Folic acid appears to be the limiting 
factor and supplementation of Schneider media with this nutrient 
improved L. (V.) braziliensis replication rates and decreased the time 
of maximum replication to 48 h.




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