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REQUEST: [ leishmaniasis ]

(12 articles match this request)



PMID: 15364993
 

TITLE: Isoenzymatic Analysis of 712 Strains of Leishmania infantum in the South
of France and Relationship of Enzymatic Polymorphism to Clinical and
Epidemiological Features.

AUTHORS: Francine Pratlong, Jean-Antoine Rioux, Pierre Marty, Françoise
Faraut-Gambarelli, Jacques Dereure, Geneviève Lanotte, Jean-Pierre Dedet

AFFILIATION: Laboratoire de Parasitologie, 163, rue Auguste Broussonet, 34090
Montpellier, France. parasito at univ-montp1.fr

REFERENCE: J Clin Microbiol 2004 Sep 42(9):4077-82

In the south of France, leishmaniasis due to Leishmania infantum occurs 
in the following five foci of endemicity (from west to east): Pyrénées
-Orientales, Cévennes, Provence, Côte d'Azur, and Corsica. Between 
1981 and 2002, 712 Leishmania strains obtained from humans, dogs, cats, 
and sand flies were studied by isoenzyme analysis. In total, seven 
zymodemes were identified: MON-1, MON-11, MON-24, MON-29, MON-33, MON-34
, and MON-108. The Pyrénées-Orientales focus is characterized by a 
predominance of human cutaneous leishmaniasis and a high enzymatic 
polymorphism (five zymodemes). In the other foci, where human visceral 
leishmaniasis is predominant, only two zymodemes are present. L. 
infantum MON-1 is the parasite most frequently found, in patients both 
with and without concomitant human immunodeficiency virus infection. MON
-1 is the only zymodeme present in dogs, which act as the reservoir host
 in all of the foci. In Cévennes, where the complete life cycle of 
zymodeme MON-1 has been identified, Phlebotomus perniciosus and 
Phlebotomus ariasi are vectors. The enzymatic polymorphism is compared 
to that of neighboring countries (Spain and Italy). In Pyrénées-
Orientales, small variant zymodemes with electromorphs of heterozygote-
like and homozygotic patterns can be explained by different genetic 
hypotheses.








PMID: 15364433
 

TITLE: Vaccination with a plasmid DNA cocktail encoding the nucleosomal histones
of Leishmania confers protection against murine cutaneous leishmaniosis.

AUTHORS: Salvador Iborra, Manuel Soto, Javier Carrión, Carlos Alonso, Jose M
Requena

AFFILIATION: Centro de Biología Molecular "Severo Ochoa", Universidad Autónoma
de Madrid, Campus de Cantoblanco, Madrid E-28049, Spain.

REFERENCE: Vaccine 2004 Sep 22(29-30):3865-76

Leishmania histones are relevant immunogens for the host immune system 
during both Leishmania infection and disease. In the present paper we 
have evaluated the prophylactic value of the four Leishmania infantum 
histones forming the nucleosomal core in the murine model of cutaneous 
leishmaniasis. In a first stage, the immune response elicited by the 
intramuscular injection of a mixture of four plasmid DNAs, encoding the 
L. infantum histones H2A, H2B, H3 and H4, was determined in BALB/c mice
. It was found that the immunized animals developed a specific Th1 
immune response, which was associated with an antigen-specific 
production of interferon (IFN-gamma) and a limited humoral response 
against histones (dominated by antibodies of the IgG2a isotype). 
According to the pure Th1-type immune response elicited by the DNA 
vaccination with Leishmania histones, vaccinated mice showed a solid 
immunity that efficiently controlled the Leishmania major infection. The
 protection in mice vaccinated with histone-DNAs was associated with a 
low humoral response against leishmanial antigens, an enhanced IFN-gamma
 production and little, if any, IL-4 production. The relative 
contribution of both CD8(+) and CD4(+) T cells to the IFN-gamma 
production, and the IL-12 dependence were also evaluated. All these data
 indicated that DNA vaccination with Leishmania histones genes results 
in a specific Th1-like response during L. major infection, and that both
 CD4(+) and CD8(+) T cells contribute to the resistance of vaccinated 
mice to cutaneous leishmaniasis.




PMID: 15364462
 

TITLE: Double-blind randomized efficacy field trial of alum precipitated
autoclaved Leishmania major vaccine mixed with BCG against canine visceral
leishmaniasis in Meshkin-Shahr district, I.R. Iran.

AUTHORS: Mehdi Mohebali, Ali Khamesipour, Iraj Mobedi, Zabih Zarei, Reza
Hashemi-Fesharki

AFFILIATION: School of Public Health and Institute of Public Health Research,
Tehran University of Medical Sciences, P.O. Box 14155-6446, Tehran, Iran.

REFERENCE: Vaccine 2004 Sep 22(29-30):4097-100

This study was designed to evaluate the efficacy of a single dose of 
aluminum hydroxide (alum) precipitated Leishmania major (Alum-ALM) 
vaccine plus BCG against canine visceral leishmaniasis. Three hundred 
and forty-seven healthy dogs with no anti-Leishmania antibodies were 
double-blind randomly injected intradermally with either 0.1ml of Alum-
ALM (200microg protein) mixed with BCG (182 dogs) or injected with 0.1ml
 of normal saline (165 dogs). The results of 16 months follow-up showed 
that the vaccine was safe and well-tolerated. Strong seroconversion 
using DAT and ELISA techniques at 16 months post-vaccination was 
considered as an indication of Leishmania infection. The incidence rate 
was 3.7% (6/162) in vaccinated group and 12.0% (17/141) in control group
 using DAT technique. The efficacy of the vaccine was calculated to be 
69.3%.




PMID: 15357081
 

TITLE: Clinical features, epidemiology, and efficacy and safety of intralesional
antimony treatment of cutaneous leishmaniasis: recent experience in Turkey.

AUTHORS: Soner Uzun, Murat Durdu, Gulnaz Culha, Adil M Allahverdiyev, Hamdi R
Memisoglu

AFFILIATION: Department of Dermatology, Cukurova University School of Medicine,
Adana 01330, Turkey. sonuzun at hotmail.com

REFERENCE: J Parasitol 2004 Aug 90(4):853-9

A total of 1,030 patients, 40.2% men and 59.8% women, identified during 
the period of October 1998 to November 2002 as having cutaneous 
leishmaniasis (CL), were studied; 1,431 lesions were identified in the 1
,030 patients. One lesion was present in 80.7% of the patients. The size
 of the lesions (longest axis) was 13.6 mm (standard, 12.1 mm; range 3-
150 mm). Most of the lesions were of the papular type (51.2%), although 
several atypical clinical presentations of CL were observed. The 
duration of the disease ranged between 1 and 72 mo (mean duration, 10.8 
mo). The clinical suspicion of CL was confirmed by the observation of 
amastigotes on lesion tissue samples stained by Giemsa. The test was 
positive in 851 of 1,030 patients (82.6%). Intralesional meglumine 
antimonate solution (85 mg Sb/ml, 0.2-1 ml, depending on the size of the
 lesion) weekly until complete cure or up to 20 wk was used for first-
line therapy of 890 patients (86.4%). We found that this regimen of 
intralesional Sb has an efficacy of 97.2% with a low relapse rate of 3.9
% and no serious adverse side effects.




PMID: 15357103
 

TITLE: Molecular characterization of the Leishmania braziliensis L6 ribosomal
protein.

AUTHORS: M C Thomas, E Martinez-Carretero, E Carmelo, A C González, B
Valladares

AFFILIATION: Instituto de Parasitologia y Biomedicina López Neyra, C.S.J.C. Av.
del Conocimiento s/n 18100 Granada, Spain.

REFERENCE: J Parasitol 2004 Aug 90(4):908-13

By screening a Leishmania braziliensis complementary DNA library with a 
pool of sera from leishmaniasis patients, the gene coding for L6 
ribosomal protein was isolated. The sequence, genomic organization, and 
transcription of this gene are described in this article. The sequence 
analysis of the L. braziliensis L6 gene shows a single open reading 
frame, which codes for a protein of 192 amino acids (aa) with a 
hypothetical molecular mass of 20.9 kDa. The protein exhibits 
significant sequence similarity to L6 ribosomal proteins from higher 
eukaryotes and yeast. Thus, the L. braziliensis L6 protein contains 4 
functional motifs, which are located at equivalent positions in other L6
 ribosomal proteins described previously. Interestingly, the L6 
ribosomal protein from L. braziliensis contains a specific region of 14 
aa and a tyrosine kinase motif, which is absent in human and C. elegans 
L6 protein. The locus coding the L. braziliensis L6 ribosomal protein is
 formed by 2 gene copies arranged in tandem and located in a chromosome 
of approximately 0.9. Mb. The genes are actively transcribed as 2 
polyadenylated transcripts of approximately 1.15 and 0.85 kb, which 
differ in their steady-state level and stability.




PMID: 15372998
 

TITLE: Case report of leishmaniasis in four cats.

AUTHORS: M G Pennisi, M Venza, S Reale, F Vitale, S Lo Giudice

AFFILIATION: Department of Medical Veterinary Sciences--Faculty of Veterinary
Medicine, University of Messina, Italy. MariaGrazia.Pennisi at unime.it

REFERENCE: Vet Res Commun 2004 Aug 28 Suppl 1():363-6




PMID: 15305319
 

TITLE: In vitro leishmanicidal activity of some scarce natural products.

AUTHORS: Marii Takahashi, Hiroyuki Fuchino, Setsuko Sekita, Motoyoshi Satake

AFFILIATION: Tsukuba Medicinal Plant Research Station, National Institute of
Health Sciences, Tsukuba, Japan.

REFERENCE: Phytother Res 2004 Jul 18(7):573-8

Leishmanicidal activity of 46 natural products including several fern 
and Betula constituents was examined. Several pterosin and atisene 
compounds from ferns had high activity. Among the triterpenoids the 
carboxyl group was found to be important for activity. In the 
diarylheptanoids, the linear-type and the diphenylether-type showed 
significant activity, and it was found that the carbonyl group at C-11 
is necessary for the activity of biphenyl-types.








PMID: 15234661
 

TITLE: The Leishmania chagasi proteasome: role in promastigotes growth and
amastigotes survival within murine macrophages.

AUTHORS: Izaltina Silva-Jardim, M Fátima Horta, F Juarez Ramalho-Pinto

AFFILIATION: Departamento de Bioquímica e Imunologia, Faculdade de Medicina de
Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, CEP
14049-900, Brazil.

REFERENCE: Acta Trop 2004 Jul 91(2):121-30

Proteasomes are multisubunit proteases that exist universally among 
eukaryotes. They have multiple proteolytic activities and are believed 
to have important roles in regulating cell cycle, selective 
intracellular proteolysis, and antigen presentation. Here we have 
partially purified Leishmania chagasi proteasome. The L. chagasi 
proteasome rich fraction displayed the typical features of eukaryotic 
20S proteasome complexes, being active towards peptidyl substrates with 
hydrophobic and acidic residues, and sensitive to the proteasome-
specific inhibitor lactacystin. We have shown that lactacystin, or its 
active form clasto-lactacystin beta-lactone, but not E-64, blocks the in
 vitro growth of L. chagasi promastigotes, demonstrating that the 
interference with parasite growth is due to the lack of proteasome 
activity. Furthermore, pre-treatment of L. chagasi promastigotes with 
lactacystin did not prevent parasite entry in host cells, but markedly 
restricted its intracellular survival. These results demonstrate that 
intact parasite proteasome function is required for replication of L. 
chagasi and for amastigotes survival inside the vertebrate host cell.




PMID: 15369643
 

TITLE: Facial and oral aspects of some venereal and tropical diseases.

AUTHORS: Marcia Ramos-E-Silva

AFFILIATION: Dermatology Sector and Post-Graduation Course University Hospital
and School of Medicine Federal University of Rio de Janeiro; Rua Sorocaba, 464
/ 205 22271-110 Rio de Janeiro Brazil; ramos.e.silva at dermato.med.br

REFERENCE: Acta Dermatovenerol Croat 2004  12(3):173-80

Diseases of the tropical areas include some venereal diseases, and they 
are still very prevalent in some countries; Brazil is one of them. Very 
few cases are originated in large cities, as Rio de Janeiro, but at the 
University Hospital of the Federal University of Rio de Janeiro we also 
see those patients who come from the interior of the State of Rio de 
Janeiro or from other states to seek medical care at better equipped 
hospitals for this type of investigation and therapy. Venereal and 
tropical dermatoses have many different cutaneous manifestations and may
 affect skin in several locations. The face is one of the affected areas
 especially when the disease has a predilection for cartilage, oral and/
or nasal mucosa. Alterations observed on the skin of the face and on the
 mucosa of the mouth of some tropical diseases, such as leprosy, 
leishmaniasis, paracoccidioidomycosis, donovanosis, and syphilis, as 
they are observed in Brazil, are presented and discussed in this article




PMID: 15267002
 

TITLE: A stable focus of canine leishmaniosis in the Bologna Province, Italy.

AUTHORS: E Mollicone, G Battelli, M Gramiccia, M Maroli, R Baldellii

AFFILIATION: Dipartimento di Sanità Pubblica Veterinaria e Patologia Animale,
Alma Mater Studiorum-Università di Bologna, via Tolara di Sopra 50, 40064
Ozzano dell'Emilia (BO), Italy.

REFERENCE: Parassitologia 2003 Jun 45(2):85-8

During an epidemiological survey carried out for two consecutive years (
2001-2002), autochthonous cases of canine leishmaniosis (CanL) were 
reported in Communes of the Bologna Province (Emilia-Romagna Region, 
northern Italy), involved in the past (1971-1972) in a severe outbreak 
of human visceral leishmaniosis (VL). Serological controls, carried out 
by immunofluorescence antibody test on a sample of owned dogs, detected 
a mean prevalence of 2.5% in the first year in 4 Communes, and of 11.2% 
in the second year in only one Commune, where an incidence value of 9.3
% was assessed. The autochthonous origin of the infection was confirmed 
in 11 out of 13 positive animals in the first year and in 5 out of 6 new
 cases in the second year. In one case the parasitological examinations 
led to the isolation of leishmaniae characterized as Leishmania infantum
 zymodeme MON 1. Entomological surveys carried out during two sandfly 
seasons in one of the areas concerned by the VL outbreak showed the 
presence of Phlebotomus perfiliewi and, to a lesser extent, of P 
perniciosus, both proven vectors of L. infantum in Italy. The results 
obtained seem to suggest the presence of a stable focus of CanL in the 
territory involved in the previous VL outbreak of 1971-1972, within 
which the infection in the canine population had been assessed only 
serologically. Such an epidemiological situation may be seen either as 
the persistence of an old focus or as a new imported one.




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PMID: 11269325
 

TITLE: Non-ulcerative cutaneous lesion in immunodeficient mice with Leishmania
amazonensis infection.

AUTHORS: M Terabe, T Kuramochi, T Hatabu, M Ito, Y Ueyama, K Katakura, S Kawazu,
T Onodera, Y Matsumoto

AFFILIATION: Department of Molecular Immunology, School of Agriculture and Life
Sciences, University of Tokyo, Japan.

REFERENCE: Parasitol Int 1999 Mar 48(1):47-53

Cutaneous leishmaniasis begins as papules or nodules at the site of 
promastigote inoculation. The next key pathogenic event in this disease 
is the formation of an ulcer at this site. Leishmania infection in 
immunodeficient mice, however, showed non-ulcerative cutaneous lesions 
suggesting the involvement of the immune system in ulcer formation. 
Severe combined immunodeficient (SCID), recombination-activating gene 2 
knockout (RAG-2-/-), and immunocompetent mice were inoculated 
subcutaneously with cultured L. amazonensis promastigotes. Macroscopic 
nodules appeared at the inoculation site within 2 weeks of infection in 
all the mice and gradually extended to the surrounding skin tissue. 
Although nodules of immunocompetent mice ulcerated within 6 weeks, 
immunodeficient mice did not form ulcers even after 25 weeks of 
inoculation. These results strongly suggest the importance of functional
 T and B cells in ulcer formation of cutaneous leishmaniasis and are 
consistent with clinical features of non-ulcerative cutaneous 
leishmaniasis in some AIDS patients. The present study also indicates 
that the L. amazonensis-infected immunodeficient mouse model might be 
suitable for studying the mechanisms of ulcer formation in cutaneous 
leishmaniasis.




PMID: 9164958
 

TITLE: Role of CD4+ T cells in pathogenesis associated with Leishmania
amazonensis infection.

AUTHORS: L Soong, C H Chang, J Sun, B J Longley, N H Ruddle, R A Flavell, D
McMahon-Pratt

AFFILIATION: Department of Epidemiology and Public Health, Yale University
School of Medicine, New Haven, CT 06520, USA.

REFERENCE: J Immunol 1997 Jun 158(11):5374-83

Most inbred strains of mice are susceptible to Leishmania amazonensis 
infection. We have examined the mechanism(s) underlying this generalized
 susceptibility using mice deficient in T cell development or in the 
expression of either MHC class I or class II. In contrast to wild-type 
C57BL/6 (B6) mice that uniformly developed large ulcerating lesions, 
mice lacking functional CD4+ T cells (due to targeted disruption of 
genes for either MHC class II trans-activator or I-A beta) showed no 
signs of lesion development for up to 12 to 14 wk postinfection and 
contained significantly lower numbers of parasites in lesions. Similarly
, both B6 nude and RAG2 -/- mice failed to develop lesions. However, 
RAG2 -/- mice reconstituted with naive wild-type CD4+ T cells and beta2m
 -/- mice did develop lesions. Lesions of MHC class II -/- mice 
contained minimal numbers of CD8+ T cells, a marked reduction of 
monocytes/macrophages, and evident extracellular parasites. The 
inability to mount an inflammatory response in MHC class II -/- mice 
correlated with the failure to produce lymphokines that lead to the 
recruitment of monocytes/granulocytes. These results demonstrate that 
CD4+ T cells are the primary lymphocyte subset that mediates cellular 
infiltration, lesion pathology, and therefore, susceptibility to L. 
amazonensis infection. The disease-promoting CD4+ T cells in L. 
amazonensis-infected mice have the characteristics of Th1 cells. The 
striking differences in the course of infection between MHC class II
 -/- mice infected with L. amazonensis and Leishmania major suggest that
 these parasites may have adapted different strategies regarding the CD4
-dependent immune response.




REQUEST: [ leishmania ]

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



PMID: 15368281
 

TITLE: Frontline: Self-peptides that bind with low affinity to the
diabetes-associated I-A(g7) molecule readily induce T cell tolerance in
non-obese diabetic mice.

AUTHORS: Walter G Ferlin, Evelyne Mougneau, Stéphanie Hugues, Heiner Appel,
Mei-Huei Jang, Julie Cazareth, Lucie Beaudoin, Corinne Schricke, Agnès Lehuen,
Kai W Wucherpfennig, Nicolas Glaichenhaus

AFFILIATION: INSERM E0344, Université de Nice-Sophia Antipolis, Valbonne,
France.

REFERENCE: Eur J Immunol 2004 Oct 34(10):2656

Although non-obese diabetic (NOD) mice spontaneously develop T cell 
autoimmunity, it is not clear whether this phenomenon results from a 
defect in tolerance to self-Ag. Furthermore, as autoimmunity has been 
postulated to result from T cell responses directed toward self-peptides
 that bind with low affinity to NOD I-A(g7) MHC class II molecules, it 
is important to determine whether the expression of such peptides 
induces tolerance. We have constructed NOD transgenic (Tg) mice 
expressing the Leishmania antigen receptor for C kinase (LACK) Ag in 
either the thymus or pancreatic beta cells. We identified LACK peptides 
that were the targets of T cells in LACK-immunized NOD mice while 
binding to I-A(g7) with low affinity. While CD4(+) T cells from NOD mice
 secreted IFN-gamma, IL-4, IL-5 and IL-10 in response to LACK, those 
from LACK-expressing Tg mice secreted reduced levels of cytokines. 
Experiments using peptide/MHC multimers showed that LACK-expressing Tg 
mice exhibited self-reactive CD4(+) T cells with impaired proliferation 
capabilities. Hence, even self-peptides that bind to I-A(g7) with low 
affinity can induce tolerance in NOD mice. This result is important in 
light of the commonly held hypothesis that T cells reacting to peptides 
that bind to MHC with low affinity escape tolerance induction and cause 
autoimmunity.




PMID: 15371479
 

TITLE: Identification of the most abundant secreted proteins from the salivary
glands of the sand fly Lutzomyia longipalpis, vector of Leishmania chagasi.

AUTHORS: Jesus G Valenzuela, Mark Garfield, Edgar D Rowton, Van M Pham

AFFILIATION: Vector Molecular Biology Unit, Laboratory of Malaria and Vector
Research, NIAID, National Institutes of Health, 12735 Twinbrook Parkway, Room
2E-22C, Rockville, MD 20852, USA.

REFERENCE: J Exp Biol 2004 Oct 207(Pt 21):3717-29

Using massive cDNA sequencing, proteomics and customized computational 
biology approaches, we have isolated and identified the most abundant 
secreted proteins from the salivary glands of the sand fly Lutzomyia 
longipalpis. Out of 550 randomly isolated clones from a full-length 
salivary gland cDNA library, we found 143 clusters or families of 
related proteins. Out of these 143 families, 35 were predicted to be 
secreted proteins. We confirmed, by Edman degradation of Lu. longipalpis
 salivary proteins, the presence of 17 proteins from this group. Full-
length sequence for 35 cDNA messages for secretory proteins is reported
, including an RGD-containing peptide, three members of the yellow-
related family of proteins, maxadilan, a PpSP15-related protein, six 
members of a family of putative anticoagulants, an antigen 5-related 
protein, a D7-related protein, a cDNA belonging to the Cimex apyrase 
family of proteins, a protein homologous to a silk protein with amino 
acid repeats resembling extracellular matrix proteins, a 5'-nucleotidase
, a peptidase, a palmitoyl-hydrolase, an endonuclease, nine novel 
peptides and four different groups of proteins with no homologies to any
 protein deposited in accessible databases. Sixteen of these proteins 
appear to be unique to sand flies. With this approach, we have tripled 
the number of isolated secretory proteins from this sand fly. Because of
 the relationship between the vertebrate host immune response to 
salivary proteins and protection to parasite infection, these proteins 
are promising markers for vector exposure and attractive targets for 
vaccine development to control Leishmania chagasi infection.








PMID: 15364110
 

TITLE: Vaccination with Leishmania soluble antigen and immunostimulatory
oligodeoxynucleotides induces specific immunity and protection against
Leishmania donovani infection.

AUTHORS: Poonam Tewary, Jayesh Mehta, Bindu Sukumaran, Rentala Madhubala

AFFILIATION: School of Life Sciences, Jawaharlal Nehru University, New Delhi
110067, India.

REFERENCE: FEMS Immunol Med Microbiol 2004 Oct 42(2):241-8

In this report, we investigated the effect of ODN containing 
immunostimulatory CG motifs as adjuvant with soluble antigen (SA) from 
Leishmania donovani. BALB/c mice were vaccinated with the soluble 
antigen with or without CpG-ODN as adjuvant and then challenged with L. 
donovani metacyclic promastigotes. CpG-ODN alone resulted in partial 
protection against challenge with L. donovani. Immunization of mice with
 SA and CpG-ODN showed enhanced reduction in parasite load ( 
approximately 60%) when compared to SA ( approximately 40%) immunized 
mice. Immunization with SA by itself resulted in a mixed Th1/Th2 
response whereas co-administration of SA with CpG-ODN resulted in a 
strong Th1 promoting isotype as they together promoted production of 
immunoglobulin G2a. Leishmania-specific Th1 cytokine response was 
induced by co-administering CpG-ODN and SA as they together promoted 
production of IFN-gamma and IL-12. In the present study, we demonstrate 
that immunostimulatory phosphorothioate-modified ODN are promising 
immune enhancers for vaccination against visceral leishmaniaisis.




PMID: 15362872
 

TITLE: Isomerase-Independent Chaperone Function of Cyclophilin Ensures
Aggregation Prevention of Adenosine Kinase Both in vitro and under in vivo
Conditions.

AUTHORS: Anutosh Chakraborty, Banibrata Sen, Rupak Datta, 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: Biochemistry 2004 Sep 43(37):11862-72

Using inactive aggregates of adenosine kinase (AdK) from Leishmania 
donovani as the model substrate, we recently demonstrated that a 
cyclophilin (LdCyP) from the same source in an isomerase-independent 
fashion reactivated the enzyme in vitro by disaggregating its inactive 
oligomers [Chakraborty et al. (2002) J. Biol. Chem. 277, 47451-47460]. 
Besides disrupting preformed aggregates, LdCyP also prevents 
reaggregation of the newly formed active protein that is generated after
 productive refolding from its urea-denatured state. To investigate 
possible physiological implications of such phenomena, a unique 
expression system that simultaneously induces both AdK and LdCyP in 
naturally AdK-deficient Escherichia coli, was developed. Both in vitro 
and in vivo experiments revealed that oligomerization is an inherent 
property of this particular enzyme. In vivo protein cross-linking 
studies, activity determination analysis and Ado phosphorylation 
experiments carried out in cells coexpressing both the proteins 
unequivocally demonstrated that, similar to the phenomena observed in 
vitro, aggregates of the enzyme formed in vivo are able to interact with
 both LdCyP and its N-terminal truncated form (N(22)(-)(88)DEL LdCyP) in
 a crowded intracellular environment, resulting in aggregation 
prevention and reactivation of the enzyme. Our results indicate that the
 isomerase-independent chaperone function of LdCyP, detected in vitro, 
participates in vivo as well to keep aggregation-prone proteins in a 
monomeric state. Furthermore, analogous to yeast/bacterial two-hybrid 
systems, development of this simple coexpression system may help in the 
confirmation of interaction of two proteins under simulated in vivo 
conditions.




PMID: 15329410
 

TITLE: A trypanothione-dependent glyoxalase I with a prokaryotic ancestry in
Leishmania major.

AUTHORS: Tim J Vickers, Neil Greig, Alan H Fairlamb

AFFILIATION: Division of Biological Chemistry and Molecular Microbiology,
Wellcome Trust Biocentre, School of Life Sciences, University of Dundee, DD1
5EH Dundee, Scotland.

REFERENCE: Proc Natl Acad Sci U S A 2004 Sep 101(36):13186-91

Glyoxalase I forms part of the glyoxalase pathway that detoxifies 
reactive aldehydes such as methylglyoxal, using the spontaneously formed
 glutathione hemithioacetal as substrate. All known eukaryotic enzymes 
contain zinc as their metal cofactor, whereas the Escherichia coli 
glyoxalase I contains nickel. Database mining and sequence analysis 
identified putative glyoxalase I genes in the eukaryotic human parasites
 Leishmania major, Leishmania infantum, and Trypanosoma cruzi, with 
highest similarity to the cyanobacterial enzymes. Characterization of 
recombinant L. major glyoxalase I showed it to be unique among the 
eukaryotic enzymes in sharing the dependence of the E. coli enzyme on 
nickel. The parasite enzyme showed little activity with glutathione 
hemithioacetal substrates but was 200-fold more active with 
hemithioacetals formed from the unique trypanosomatid thiol 
trypanothione. L. major glyoxalase I also was insensitive to glutathione
 derivatives that are potent inhibitors of all other characterized 
glyoxalase I enzymes. This substrate specificity is distinct from that 
of the human enzyme and is reflected in the modification in the L. major
 sequence of a region of the human protein that interacts with the 
glycyl-carboxyl moiety of glutathione, a group that is conjugated to 
spermidine in trypanothione. This trypanothione-dependent glyoxalase I 
is therefore an attractive focus for additional biochemical and genetic 
investigation as a possible target for rational drug design.




PMID: 15372110
 

TITLE: A serine/threonine kinase, Cot/Tpl2, modulates bacterial DNA-induced
IL-12 production and Th cell differentiation.

AUTHORS: Kenji Sugimoto, Mutsuhiro Ohata, Jun Miyoshi, Hiroyoshi Ishizaki,
Naotake Tsuboi, Akio Masuda, Yasunobu Yoshikai, Masaya Takamoto, Kazuo Sugane,
Seiichi Matsuo, Yasuhiro Shimada, Tetsuya Matsuguchi

AFFILIATION: Division of Host Defense, Center for Neural Disease and Cancer,
and.

REFERENCE: J Clin Invest 2004 Sep 114(6):857-66

A serine/threonine protein kinase, Cot/Tpl2, is indispensable for 
extracellular signal-regulated kinase (ERK) activation and production of
 TNF-alpha and PGE(2) in LPS-stimulated macrophages. We show here that 
Cot/Tpl2 is also activated by other Toll-like receptor (TLR) ligands. 
Bacterial DNA rich in the dinucleotide CG (CpG-DNA), unlike LPS or 
synthetic lipopeptide, activated ERK in a Cot/Tpl2-independent manner. 
Peritoneal macrophages and bone marrow-derived DCs from Cot/Tpl2(-/-) 
mice produced significantly more IL-12 in response to CpG-DNA than those
 from WT mice. Enhanced IL-12 production in Cot/Tpl2(-/-) macrophages is
, at least partly, regulated at the transcriptional level, and the 
elevated IL-12 mRNA level in Cot/Tpl2(-/-) macrophages is accompanied by
 decreased amounts of IL-12 repressors, such as c-musculoaponeurotic 
fibrosarcoma (c-Maf) and GATA sequence in the IL-12 promoter-binding 
protein (GA-12-binding protein; GAP-12) in the nucleus. Consistently, 
Cot/Tpl2(-/-) mice showed Th1-skewed antigen-specific immune responses 
upon OVA immunization and Leishmania major infection in vivo. These 
results indicate that Cot/Tpl2 is an important negative regulator of Th1
-type adaptive immunity, that it achieves this regulation by inhibiting 
IL-12 production from accessory cells, and that it might be a potential 
target molecule in CpG-DNA-guided vaccination.




PMID: 15372990
 

TITLE: Haemostatic disorders in dogs naturally infected by Leishmania infantum.

AUTHORS: M Corona, P Ciaramella, A Pelagalli, L Cortese, M E Pero, D Santoro, P
Lombardi

AFFILIATION: Department of Veterinary Clinical Science--Section of Internal
Medicine, University of Naples, Italy.

REFERENCE: Vet Res Commun 2004 Aug 28 Suppl 1():331-4




PMID: 15368649
 

TITLE: Anthranoid compounds with antiprotozoal activity from Vismia orientalis.

AUTHORS: Zakaria H Mbwambo, Sandra Apers, Mainen J Moshi, Modest C Kapingu,
Sabine Van Miert, Magda Claeys, Reto Brun, Paul Cos, Luc Pieters, Arnold
Vlietinck

AFFILIATION: Institute of Traditional Medicine, Muhimbili University College of
Health Sciences, Dar es Salaam, Tanzania. zmbwambo at muchs.ac.tz

REFERENCE: Planta Med 2004 Aug 70(8):706-10

A phytochemical investigation of the 80% ethanolic extract of stem bark 
of Vismia orientalis Engl. (Guttiferae or Clusiaceae), a plant used in 
traditional medicine in Tanzania, resulted in the isolation and 
spectroscopic characterisation of 3-geranyloxy-6-methyl-1,8-
dihydroxyanthraquinone, emodin, vismione D and bianthrone A1. Vismione D
 exhibited a broad range of antiprotozoal activities against Trypanosoma
 brucei rhodesiense and T. cruzi (IC50 < 10 micrograms/mL), 
Leishmania donovani (IC50 0.37 micrograms/mL) and Plasmodium falciparum 
strain K1 (IC50 1.0 microgram/mL). However, it was also slightly 
cytotoxic against human L6 cells (IC50 4.1 micrograms/mL). Emodin showed
 antileishmanial activity (IC50 2.0 micrograms/mL), while its IC50 
against L6 cells was 20.3 micrograms/mL. Other antiprotozoal activities 
observed for emodin against both Trypanosoma species and P. falciparum, 
for bianthrone A1 against T. b. rhodesiense and P. falciparum, and for 3
-geranyloxy-6-methyl-1,8-dihydroxyanthraquinone against T. b. 
rhodesiense, L. donovani and P. falciparum were in the range of 10 to 50
 micrograms/mL. None of the compounds showed antibacterial or antiviral
 (including also HIV) activity.








PMID: 15372989
 

TITLE: Plasma thrombomodulin levels in dogs naturally infected with Leishmania
infantum.

AUTHORS: P Ciaramella, L Cortese, M Corona, R Ambrosio, A Di Loria, A
Persechino

AFFILIATION: Department of Veterinary Clinical Science--Section of Internal
Medicine--University of Naples Federico II, via Delpino, 1--80137 Naples,
Italy. paociara at unina.it

REFERENCE: Vet Res Commun 2004 Aug 28 Suppl 1():327-30




PMID: 15234664
 

TITLE: Comparison of the effectiveness of two topical paromomycin treatments
versus meglumine antimoniate for New World cutaneous leishmaniasis.

AUTHORS: Rodrigo X Armijos, M Margaret Weigel, Manuel Calvopiña, Manuel
Mancheno, Roberto Rodriguez

AFFILIATION: Health Sciences Program, College of Health Sciences, Room 705, 1101
North Campbell Street, The University of Texas at El Paso, El Paso, TX
79902-0581, USA.

REFERENCE: Acta Trop 2004 Jul 91(2):153-60

The randomized, controlled study compared the therapeutic efficacy and 
safety of two paromomycin-containing topical preparations with the gold 
treatment standard, meglumine antimoniate, and with each other in 120 
Ecuadorian patients with ulcerated lesions. The two paromomycin 
treatment comparisons were double-blinded. Group 1 (n = 14) received 15
% paromomycin plus 12% methylbenzonium chloride (PR-MBCL) dissolved in a
 soft white paraffin base, applied twice daily for 30 days. Group 2 (n
 = 40) was also treated for 30 days with 15% paromomycin plus 10% urea (
PR-U) dissolved in the same paraffin base. Group 3 (n = 40) received 
20mg/kg/day of IM meglumine antimoniate (MA) for 10 days as per 
Ecuadorian Ministry of Public Health recommendations at the time of the 
study. The 10-day treatment was completed by 90% of the MA group 
compared to 72.5% of the PR-MBCL (X2 = 4.0, P = 0.045) and 75% of the PM
-U (X2 = 3.1, P > 0.05) groups whose treatment regime lasted 20 days 
longer than the MA treatment. Post-treatment lesion burning, redness, 
inflammation, and soreness were more common in the two paromomycin 
groups compared to MA group (P < 0.05). The frequency of treatment-
related side effects in the two paromomycin groups was similar. Six 
weeks after the start of treatment, 80.6% of MA subjects were clinically
 cured compared to 48.3% in the PR-MBCL (X2 = 6.1, P = 0.014) and 40% in
 the PM-U groups (X2 = 12.6, P = 0.002). By 12 weeks, the proportion of 
clinically cured subjects in the MA (91.7%) compared to PM-MBCL (79.3%) 
or PM-U (70%) groups was not significantly different (P > 0.05). MA-
treated subjects clinically cured by 12 weeks had a faster mean healing 
time (29.5 +/- 12.2 days) compared to those in the PM-MBCL (versus 43.1
 +/- 14.4 days, t = -3.7, P = 0.001) or PR-U groups (43.5 +/- 17 days; t
 = -3.2, P = 0.002). During the 48-week post-treatment follow-up period
, infection reactivation was observed in 15.2% of the MA subjects 
compared to 17.4% in the PM-MBCL and 10.5% PM-U of subjects diagnosed as
 clinically healed by 12 weeks (P > 0.05). The results suggest that 
although the time required for the clinical healing of ulcerated lesions
 takes longer, topical paromomycin may be an acceptable therapeutic 
alternative in endemic areas where meglumine antimoniate is not 
available, is too costly or medically contraindicated.




PMID: 15363945
 

TITLE: The modelling of mononuclear phagocyte-connective tissue adhesion in
vitro: application to disclose a specific inhibitory effect of Leishmania
infection.

AUTHORS: Djalma G F Carvalhal, Aryon Barbosa, Micely D'El-Rei Hermida, Jorge
Clarencio, Nathanael F Pinheiro, Patricia S T Veras, Washington L C Dos-Santos

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

REFERENCE: Exp Parasitol 2004 Jul-Aug 107(3-4):189-99

In this work, we have developed an adhesion assay to study interactions 
between mononuclear phagocytes and connective tissue in vitro and show 
its potential use to study diseases caused by intracellular 
microorganisms. The assay reproduces most of the characteristics of 
macrophage adhesion to connective tissue in vivo, such as: preferential 
adhesion to inflamed connective tissue, divalent cation and integrin 
dependence, and up-regulation upon cell activation. The phagocyte 
adhesion to connective tissue was inhibited by infection with Leishmania
 (58+/-22%, [Formula: see text] ) and was not affected by infection with
 Mycobacterium or by endocytosis of latex beads. Manganese partially 
reverted the loss in adherence produced by Leishmania infection, 
indicating that the mechanisms regulating the function of integrins are 
affected by cell infection with Leishmania. This assay might be a useful
 tool for the study of the mechanisms by which mononuclear phagocytes 
play a role in the immune-inflammatory response and in the development 
of lesions. Index Descriptors and Abbreviations: Adhesion assay; Cell 
adhesion; Macrophage adhesion; J774 cells; Leishmania amazonensis; 
Leishmania braziliensis; Leishmania chagasi; Mycobacterium fortuitum; 
Promastigotes; ANOVA, analysis of variance; CD, cluster designation for 
classification of cell markers; CS-1, connecting segment 1 of 
fibronectin; EDTA, ethylenediaminetetracetic acid; FACS, fluorescence 
activated cell sorting; FBS, fetal bovine serum; HBSS, Hanks' balanced 
salt solution; LFA-1, leukocyte activation antigen 1 (beta2-integrin, 
CD11a/CD18); LPS, lypopolysaccharide; Mac-1, beta2-integrin named after 
macrophages (CD11b/CD18); PBS, phosphate-buffered saline; RGD, peptide 
containing RGD sequence; RPMI, Roswell Park Memorial Institute-1640 (
tissue culture medium); SNK, Student-Newman-Keuls test; VCAM-1, vascular
 cell adhesion molecule 1




PMID: 15363943
 

TITLE: Leishmania (Leishmania) amazonensis: purification and characterization of
a promastigote serine protease.

AUTHORS: Raquel Elisa Da Silva-Lopez, Salvatore Giovanni-De-Simone

AFFILIATION: Laboratório de Bioquímica de Proteínas e Peptídeos,
Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz,
FIOCRUZ, Av. Brasil 4365, 21045-900, Rio de Janeiro, RJ, Brazil.

REFERENCE: Exp Parasitol 2004 Jul-Aug 107(3-4):173-82

Pathogenic protozoan proteases play crucial roles in the host-parasite 
interaction, and its characterization contributes to the understanding 
of protozoan disease mechanisms. A Leishmania amazonensis promastigote 
protease was purified 36-fold, using aprotinin-agarose affinity 
chromatography and gel filtration high performance liquid chromatography
, yielding a total recovery of 49%. The molecular mass of active enzyme 
obtained from native gel filtration HPLC and SDS-PAGE under conditions 
of reduction and non-reduction was 68kDa, suggesting that the enzyme may
 exist as a monomer. The protease isoelectric point (pI) was around 4.45
 and, as demonstrated by deglycosylation assay, it did not have any 
carbohydrate content. The optimal pH and temperature of the enzyme were 
8.0 and 28 degrees C, respectively, determined using alpha-N-rho-tosyl-l
-arginyl-methyl ester (l-TAME) as substrate. Assays of thermal stability
 indicated that 50% of the enzymatic activity was preserved after 4min 
of pre-treatment at 42 degrees C and after 24h of pre-treatment at 37 
degrees C, both in the absence of substrate. Hemoglobin, bovine serum 
albumin (BSA), ovalbumin, and both gelatin and peptide substrates 
containing arginine in ester bound were hydrolyzed by 68kDa protease. 
The insulin beta-chain was also hydrolyzed by the protease, and four 
peptidic bonds (L(11)-V(12), E(13)-A(14), L(15)-Y(16), and Y(16)-L(17)) 
were susceptible to the 68-kDa protease action. Inhibition studies 
suggested that the enzyme belonged to a serine protease class inhibited 
by calcium ions and activated by manganese ions. These findings 
demonstrate that the L. amazonensis 68-kDa serine protease differs from 
those of other protozoan parasites.




********************************************************************************************************************

 The following references are revised files and are brought to you in accordance
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********************************************************************************************************************


PMID: 12682728
 

TITLE: Characterization of the human autoimmune response to the major C-terminal
epitope of the ribosomal P proteins.

AUTHORS: M Mahler, K Kessenbrock, J Raats, R Williams, M J Fritzler, M
Blüthner

AFFILIATION: Pharmacia Diagnostics, Munzingerstrasse 7, 79111 Freiburg, Germany.
m.mahler.job at web.de

REFERENCE: J Mol Med 2003 Mar 81(3):194-204

Autoantibodies to the ribosomal phospho (-P) proteins P0, P1, and P2, 
collectively referred to as Rib-P, are specifically found in 10-40% of 
patients with systemic lupus erythematosus (SLE). These antibodies are 
believed to be correlated with lupus nephritis, hepatitis, and central 
nervous system involvement. The major immunoreactive epitope of these 
ribosomal antigens has been localized to the carboxy terminus, which is 
a highly conserved domain of all three proteins and contains two 
phosphorylated serine residues. The phosphorylated amino acids of the P 
proteins are known not to be critical epitope determinants. Furthermore
, epitope-mapping studies have shown that the major epitope is located 
within the last 11 C-terminal amino acids. Using peptide arrays we 
identified more precisely this shared epitope as the six C-terminal 
amino acids GFGLFD and elucidated the molecular recognition events of 
anti-Rib-P antibodies at the amino acid level. We identified Phe(111) 
and Phe(114) of Rib-P2 as the key residues for the interaction, with 
further contributions of Gly-112 and Asp-115. This amino acid stretch is
 also present in proteins of several pathogenic micro-organisms such as 
Trypanosoma cruzi, Brugia malayi, Pseudomonas aeruginosa, Candida 
albicans, several Leishmania species, and Bartonella henselae. Using 
newly developed ELISA systems with a C-terminal peptide (C22) and the 
recombinant proteins (P0, P1, and P2) as antigens we found a high 
specificity of anti-Rib-P antibodies for SLE and demonstrated positive 
correlations with anti-U1-C, anti-Sm-B/B' and anti-D and anti-dsDNA 
antibodies. The sensitivity and specificity in the peptide (C22) based 
assay varied between 12.8%/100% and 23.4%/96.7% for SLE, depending on 
the assigned cutoff. In contrast to other studies, we found no 
significant correlation of anti-Rib-P reactivity with central nervous 
system manifestations or renal involvement in SLE patients. We conclude 
that the epitope motif GFGLFD in the C-termini of the ribosomal P 
proteins is the key determinant of anti-Rib-P antibodies, and that the 
C22 peptide and the recombinant proteins can be used equally well for 
the detection of anti-Rib-P antibodies. The role of the major Rib-P 
epitope in the development of anti-ribosomal P antibodies and in the 
pathogenesis of SLE remains a subject of further investigation.




PMID: 9476797
 

TITLE: Identification of a transcription factor like protein at the TOR locus in
Leishmania mexicana amazonensis.

AUTHORS: S Detke

AFFILIATION: University of North Dakota School of Medicine, Department of
Biochemistry and Molecular Biology, Grand Forks 58202, USA.
sdetke at mail.med.und.nodak.edu

REFERENCE: Mol Biochem Parasitol 1997 Dec 90(2):505-11

The TOR gene (TOxic nucleoside Resistance gene) was mapped to a 2.3 kb 
fragment on the amplified DNA from tubercidin resistant Leishmania (TUB
). This DNA fragment conferred upon wild type cells resistance to 
tubercidin, inosine dialdehyde, formycin A and B and allopurinol 
riboside and a reduced ability to accumulate purine nucleobases and 
nucleosides. These properties were characteristic of the parental TUB 
cells which carried the intact amplified DNA and have been hypothesized 
to be caused by a reduction in the activity of the multiple purine 
transporters within this organism. The TOR gene was found to be 
partially homologous to the rodent and human Oct-6/SCIP/Tst-1 gene. It 
lacked, however, the POU specific domain of this class of transcription 
factors and contained only the first two helices of the POU homeodomain
. This truncated homeodomain was not required to confer resistance upon 
wild type cells to toxic nucleosides, suggesting that TOR was not a 
repressor with independent DNA binding capability.




PMID: 8619813
 

TITLE: Cloning and characterization of a ribosomal P protein from Taenia solium,
the aetiological agent of human cysticercosis.

AUTHORS: B H Kalinna, D P McManus

AFFILIATION: Molecular Parasitology Unit, Queensland Institute of Medical
Research, Bancroft Centre, Brisbane, Australia. berndK at qimr.edu.au

REFERENCE: Biochem Biophys Res Commun 1996 Feb 219(1):231-7

A cDNA encoding the complete open reading frame of the Taenia solium 
acidic ribosomal phosphoprotein P2 has been cloned. The 417 bp cDNA (
arpP2) was isolated from a T. solium cDNA expression library by PCR 
using P protein specific pimers. The ORF encodes a protein of 121 amino 
acids exhibiting 40-55% identity to mammalian, fungal, insect, and 
parasite P2 proteins. The inferred molecular mass of the protein is 12,
592 Daltons, similar to that reported for other ribosomal P2 proteins. 
After subcloning and expression, the recombinant protein was purified by
 affinity chromatography under non-denaturing conditions and was shown 
to have a molecular mass of 15 kDa which is equivalent to the expected 
size of the full length recombinant fusion protein, comprising the 12.5 
kDa arpP2 plus an additional 2 kDa for the N-terminal fusion peptide 
incorporating the six histidine residues required for purification.




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