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1.
Arch Razi Inst ; 78(6): 1822-1835, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38828174

RESUMO

Snake venoms are rich in valuable substances that have medical potential in the diagnosis and treatment of hemostatic diseases. The present paper was aimed at the purification and functional characterization basis of a thrombin-like enzyme and its role in the functioning of the coagulation cascade and platelet aggregation pathway. A thrombin-like serine protease was purified from the Iranian Echis carinatus venom (TLIECV), employing a one-step chromatographic procedure. This peptide was collected in high yield and purity by a single chromatographic step using RP-HPLC equipped with a C18 column. This peptide showed a 3000 Da molecular weight in gel-electrophoresis. Evidence in the SDS-PAGE gel has confirmed high recovery of fraction in optimal terms. Subsequently, this peptide was identified via its intact molecular mass and peptide mass fingerprint (PMF) using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Multiple sequence alignments were performed by ClustalW, the Bioedit software. Molegro Data Modeller (MDM) 3.0 software was used to predict the putative tertiary structure of the peptide. The enzyme possessed fibrinogenolytic, procoagulant, and aggregation inducer properties. Moreover, the SDS-PAGE (12%) was applied to examine fibrinogenolytic function. The purified enzyme degraded the Aα chain of fibrinogen while the Bß and γ chains were not digested. According to that, the deficient human plasma in factor X and normal human plasma were also coagulated by TLIECV, it takes part in the common and intrinsic routes of the coagulation cascade. These findings proved that TLIECV is a serine protease identical to procoagulant thrombin-like snake venom proteases; however, it specifically releases the Aα chain of bovine fibrinogen. Because of its function to make up for the deficiency of factor X and its platelet aggregation inducer property, TLIECV could be considered a molecular impact to reveal the hemostasis mechanisms.


Assuntos
Venenos de Víboras , Viperidae , Animais , Venenos de Víboras/enzimologia , Venenos de Víboras/química , Irã (Geográfico) , Trombina , Agregação Plaquetária/efeitos dos fármacos , Humanos , Eletroforese em Gel de Poliacrilamida , Sequência de Aminoácidos , Echis , Serpentes Peçonhentas
2.
Arch Razi Inst ; 72(3): 159-164, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-30341936

RESUMO

Contagious agalactia (CA) is a highly infectious disease of goats and sheep, and is a form of Mycoplasmosis, which is usually enzootic. Since Mycoplasma agalactiae (M. agalactiae) is the main cause of this disease in goats, the aim of this study was to isolate and detect M. agalactiae from semen of goat bucks. Thirty-nine semen samples were collected from goat bulks, and all samples were cultured in PPLO broth medium supplemented for M. agalaciae isolation. The bacteria DNAs were extracted from clinical samples and the PCR assay was applied to detect Mycoplasma genus and M. agalactiae species using specific primers, which amplified a 163bp fragment in 16SrRNA gene and a 375bp fragment in lipoprotein gene. The PCR evaluations were performed for both the clinical samples and the cultures. Out of the 39 samples, 29 (74.3%) of the cultures were shown positive and typical Mycoplasma colonies grew on PPLO agar, which could be considered as the diagnostic method. In addition, 38 (97.4%) samples had positive PCR results for Mycoplasma genus and six (15.3%) of the samples were shown to be positive using PCR for M. agalactiae as the diagnostic method. In the present study, M. agalactiae was detected in semen of goat bulks for the first time in Iran. Therefore, it is recommended to concern semen as one of the significant sources for this pathogen and the possibility for transmission to the female goats through semen is highlighted. Moreover, presence of this microorganism in semen could be involved in infertility of goat population.


Assuntos
Doenças das Cabras/diagnóstico , Doenças das Cabras/epidemiologia , Infecções por Mycoplasma/veterinária , Mycoplasma agalactiae/isolamento & purificação , Sêmen/microbiologia , Animais , Doenças das Cabras/microbiologia , Cabras , Irã (Geográfico)/epidemiologia , Masculino , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/epidemiologia , Infecções por Mycoplasma/microbiologia , Reação em Cadeia da Polimerase/veterinária , Prevalência
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