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Potential fibrinolytic activity of an endophytic Lasiodiplodia pseudotheobromae species.

ABSTRACT: Cost-effective, fibrinolytic agents possessing least or no antigenic properties are much in demand for their prospective use as clot busters in thrombolytic therapy. The present communication explores the potential of 22 endophytic Botryosphaeria species isolated from stem of Aegle marmelos for their fibrinolytic potential. Only nine fungal isolates exhibited proteolytic activity out of which only four possessed plasminogen-independent fibrinolytic activity. The endophytic fungal isolate #1088 AMSTITYEL showed maximum in vitro proteolytic and fibrinolytic activity with a halo formation of 153.86 and 113.04 mm², respectively. The partially purified protein exhibited a fibrinolytic activity equivalent to 6.51 U/ml of plasmin. SDS-PAGE of the dialysed fraction of #1088 AMSTITYEL resolved into six bands ranging from 26 to 80 kDa. Fibrin zymogram exhibited that a single band of molecular size ~80 KDa possessing the fibrinolytic activity. Furthermore, the bioactive isolate #1088 AMSTITYEL was identified as Lasiodiplodia pseudotheobromae using classical and molecular taxonomic tools.

SUBMITTER: Meshram V

PROVIDER: S-EPMC5398192 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature

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Potential fibrinolytic activity of an endophytic Lasiodiplodia pseudotheobromae species.

Meshram Vineet V Saxena Sanjai S

3 Biotech 20160519 1

Cost-effective, fibrinolytic agents possessing least or no antigenic properties are much in demand for their prospective use as clot busters in thrombolytic therapy. The present communication explores the potential of 22 endophytic Botryosphaeria species isolated from stem of Aegle marmelos for their fibrinolytic potential. Only nine fungal isolates exhibited proteolytic activity out of which only four possessed plasminogen-independent fibrinolytic activity. The endophytic fungal isolate #1088 A ...[more]

PMID: 28330184

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Project description:BackgroundPostpartum hemorrhage (PPH) is still the leading cause of maternal morbidity and mortality worldwide. While impaired fibrin polymerization plays a crucial role in the development and progress of PPH, recent approaches using viscoelastic measurements have failed to sensitively detect early changes in fibrinolysis in PPH. This study aimed to evaluate whether women experiencing PPH show alterations in POC-VET fibrinolytic potential during childbirth and whether fibrinolytic potential offers benefits in the prediction and treatment of PPH.MethodsBlood samples were collected at three different timepoints: T0 = hospital admission (19 h ± 18 h prepartum), T1 = 30-60 min after placental separation, and T2 = first day postpartum (19 h ± 6 h postpartum). In addition to standard laboratory tests, whole-blood impedance aggregometry (Multiplate) and viscoelastic testing (VET) were performed using the ClotPro system, which included the TPA-test lysis time, to assess the POC-VET fibrinolytic potential, and selected coagulation factors were measured. The results were correlated with blood loss and clinical outcome markers. Severe PPH was defined as a hemoglobin drop > 4g/dl and/or the occurrence of shock or the need for red blood cell transfusion.ResultsBlood samples of 217 parturient women were analyzed between June 2020 and December 2020 at Heidelberg University Women's Hospital, and 206 measurements were eligible for the final analysis. Women experiencing severe PPH showed increased fibrinolytic potential already at the time of hospital admission. When compared to non-PPH, the difference persisted 30-60 min after placental separation. A higher fibrinolytic potential was accompanied by a greater drop in fibrinogen and higher d-dimer values after placental separation. While 70% of women experiencing severe PPH showed fibrinolytic potential, 54% of those without PPH showed increased fibrinolytic potential as well.ConclusionWe were able to show that antepartal and peripartal fibrinolytic potential was elevated in women experiencing severe PPH. However, several women showed high fibrinolytic potential but lacked clinical signs of PPH. The findings indicate that high fibrinolytic potential is a risk factor for the development of coagulopathy, but further conditions are required to cause PPH.

Project description:Lasiodiplodia theobromae, a grapevine trunk pathogen, is becoming a significant threat to vineyards worldwide. In Peru, it is responsible for Botryosphaeria dieback in many grapevine-growing areas and it has spread rapidly due to its high transmissibility; hence, control measures are urgent. It is known that some endophytic bacteria are strong inhibitors of phytopathogens because they produce a wide range of antimicrobial molecules. However, studies of antimicrobial features from endophytic bacteria are limited to traditional confrontation methods. In this study, a MALDI mass spectrometry-based approach was performed to identify and characterize the antifungal molecules from Bacillus velezensis M1 and Bacillus amyloliquefaciens M2 grapevine endophytic strains. Solid medium antagonism assays were performed confronting B. velezensis M1 - L. theobromae and B. amyloliquefaciens M2 - L. theobromae for antifungal lipopeptides identification. By a MALDI TOF MS it was possible identify mass spectra for fengycin, iturin and surfactin protoned isoforms. Masses spectrums for mycobacillin and mycosubtilin were also identified. Using MALDI Imaging MS we were able to visualize and relate lipopeptides mass spectra of fengycin (1463.9 m/z) and mycobacillin (1529.6 m/z) in the interaction zone during confrontations. The presence of lipopeptides-synthesis genes was confirmed by PCR. Liquid medium antagonism assays were performed for a proteomic analysis during the confrontation of B. velezensis M1 - L. theobromae. Different peptide sequences corresponding to many antifungal proteins and enzymes were identified by MALDI TOF MS/MS. Oxalate decarboxylase bacisubin and flagellin, reported as antifungal proteins, were identified at 99 % identity through peptide mapping. MALDI mass spectrometry-based identification of antifungal molecules would allow the early selection of endophytic bacteria with antifungal features. This omics tool could lead to measures for prevention of grapevine diseases and other economically important crops in Peru.

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Potential fibrinolytic activity of an endophytic Lasiodiplodia pseudotheobromae species.

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Potential fibrinolytic activity of an endophytic Lasiodiplodia pseudotheobromae species.

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