Introduction 7 Visceral Leishmaniasis( VL )is characterized by a spectrum of
clinical features, with high cure rates in early diagnosis.
Objective: the objective is to describe the clinical and epidemiological features and
laboratory variables of
children with visceral leishmaniasis ,And to define the importance
of a positive direct investigation of the parasite in Bone marrow in the diagnosis of disease
.
Patients and Methods : It was a case series study of )52( cases of Visceral
Leishmaniasis which were hospitalized between June-20 31and June-2014 in Al -Assad
University Hospital ,Latakia
Results 52 7 7 children were included in this study .Majority of the patients (92%)
presented with fever ,common findings in physical examination were(100%) Pallor and
(97%) splenomegaly
The direct investigation of parasite in Bone marrow was positive in )%55( of cases .
The leishmaniasis is a serious health problem in Syria, due to the
wide spreading of Leishmania tropica parasite , the difficulty of
controlling the reservoirs of parasites, and there is no specific ,safe
and active therapy .Due to lack of studies
of Leishmania tropica
genome , as initial step we detect the presence of SW3 gene.
In our study, the
presence and expression of P27 gene in L.tropica evaluated in
promastigotes by PCR and RT-PCR using specific primer pairs
that manually designed after determine the consensus sequence of
forward and reverse region of primers bet
ween species of
Leishmania by using MUSCLE (Multiple Sequence Comparison by
Log- Expectation) tool for alignment. The results proved the
presence of P27 gene in Leishmania tropica, beside presence the
mRNA of P27 gene that confirm that P27 expressed in
promastigote form of L.tropica
DNA was extraction from scanning microscopic slides to
identification leishmaniases. Three methods used to DNA
extraction, first one using ready extraction solvent (Promega kit),
second depended on phenol-chloroform-iso propanol , whereas third
one depended on columns with silica gel membranes (Qia gen kit).
The results showed that (Promega kit) method didn’t gave high
concentration of DNA. (Qia gen kit) method was the best for DNA
concentration and purity
the leishmaniasis is a serious health problem in Syria, due to the
wide spreading of Leishmania Tropica parasite , the hardness of
controlling the reservoirs of parasites, and there is no specific ,safe
active therapy. A Heat shock protein 20 HSP2
0 belongs to the
small HSP (sHSP) family.
The aim of this work was determining the role of
Protein kinase C (PKC) signaling pathway in apoptosis of
Leishmania tropica promastigotes, through studying effects of PKC
specific inhibitor Staurosporine (STS) on the parasite. Our results
showed
that treated L. tropica with Staurosporine was
enough to inhibit PKC pathway completely, while viability test
showed that the concentration of STS to inhibit cell growth by 50%
(IC50 = 2.146μM).
Kinetoplastid membrane protein-11 (KMP-11), a protein present in all kinetoplastid
protozoa studied up to date, is considered a potential vaccine candidate for Leishmaniasis.
Such vaccine molecules must be expressed in amastigotes which represent t
he infective
forms for mammals, while promastigotes are the flagellate forms found in the insect hosts.
However, the expression of KMP-11 in amastigotes is still a subject of controversy. In this
study, a strain of L. tropica was isolated, cultivated, and genotyped. The expression of
KMP-11 gene in this strain was evaluated in promastigotes and in amastigotes by RT-PCR
using specific primer pairs. The results proved the presence of mRNA of KMP-11 in both
promastigotes and amastigotes forms of L. tropica. The expression of this molecule in
amastigotes is consistent with the previously demonstrated immunoprotective capacity of
KMP-11 DNA vaccine as well as the presence of humoral and cellular immune responses
against KMP-11 in Leishmania-infected animals.
Several strains of leishmania were isolated from patients suffering
from cutaneous leishmaniasis in Damascus countryside; and cultured in
vitro on semisolid medium (agar with rabbits blood) at first, then in
monophasic RPMI-١٦٤٠ medium supplemente
d with ١٠٪ (v/v) heatinactivated
fetal calf serum to determine the ideal conditions of culture.
The types of these strains were specified as Leishmania major using
isoenzymatic electrophoresis.
