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Register a new userThe inhibition role of protein kinase C (PKC) in programmed cell death of Leishmania tropica promastigotes
دور البروتين كيناز (C (PKC في تثبيط الموت الخلوي المبرمج للأشكال أمامية السوط لطفيلي الليشمانيا المدارية
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يهدف هذا العمل إلى التحري عن دور السبيل الخلوي الخاص بالبروتين كيناز PKC) C) في الموت الخلوي المبرمج للأشكال أمامية السوط لطفيلي الليشمانيا المدارية، من خلال دراسة تأثير مادة مثبطة لهذا السبيل و هي STS) Staurosporine). أظهرت النتائج أن معالجة الأشكال أمامية السوط المستنبتة من المثبط كافية لكبح السبيل الإشاري ل PKC بشكل كلي، بينما أظهر اختبار السمية أن تركيز المثبط القاتل للنصف كان (IC50 = 2.146μM).
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).
References used
Steller, H., 1995. Mechanisms and genes of cellular suicide. Science, 267(5203): p. 1445-9
Vaux, D.L., 1993. Toward an understanding of the molecular mechanisms of physiological cell death. Proc Natl Acad Sci U S A, 90(3): p. 786-9
Vaux, D.L. and A. Strasser, 1996. The molecular biology of apoptosis. Proc Natl Acad Sci U S A, 93(6): p. 2239-44
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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.
The parasitic antigens are of great interest in immune responses and in the
strategies for developing vaccines. The aim of this study was to identify the
major antigens which are expressed in the two leishmania tropica stages,
promastigote & amast
igote, employing the western blot analysis using the sera
collected from mice immunized with the above mentioned parasite stages' total
protein.
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
Leishmaniasis spreads in eighty-eight countries, including the Syrian Arab
Republic. This disease is caused by a protozoic parasite of the genus
Leishmania. Twenty-two species of leishmania were reported to be pathogenic
for human. The disease is
presented in three clinical forms: cutaneous,
mucocutaneous and visceral leishmaniasis. A set of methods for diagnosis and
identifying the type of parasite have been developed in order to give
appropriate treatment. These methods include isoenzyme analysis, serological
and immunological methods and DNA hybridization. The polymerase chain
reaction (PCR), using genomic or kinetoplastid DNA; provides an excellent tool
for diagnosis and characterization of leishmania species. In this work, the two
types of DNA were extracted in one step from cultured isolates taken from
patients. We have used kinetoplastid DNA and specific primers to determine
parasite species by PCR. These primers amplify species specific fragment. In
this study, we have determined that the causative species of the cutaneous
leishmaniasis in all patients was L.tropica.
We studied a population of 47 patients who were admitted to Alassad Hospital in
Latakia with Community-Acquired Pneumonia from April 2013 to April 2014 , 21 women
and 26 men, they were followed up every 48 hours over eight days.
Patients were divi
ded into three groups according to clinical stability criteria, and we
studied the changes in CRP values at each group of patients separately and found the
following:
RapidResponse 25.5%: the value of CRP decreased on the fourth day by 48.1%
SlowResponse 38.3%: the value of CRP in the fourth day decreased by 14.1% but fell on
the sixth day by 35.7%TreatmentFailure 36.2% : the value of CRP have not fallen on the
fourth day, but became 103%. on the sixth day decreased slightly by 5.9%, on the eighth
day fell by 39%.
Conclusion: CRP changes Coincided with the patients' response to treatment well so
they can be relied upon (In addition to the other stability criteria but not alone) in
determining the type of response and thus making a decision about the treatment plan.
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