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Evaluating the Antimicrobial and Anti-Hemolytic Activity of Synthesized Pseudopeptide against Leptospiral Species: In Silico and In Vitro Approach

Bacterial infections are one of the leading causes of morbidity, mortality, and healthcare complications in patients. Leptospirosis is found to be the most prevalent, re-emergent, and neglected tropical zoonotic disease worldwide. The adaptation to various environmental conditions has made Leptospir...

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Autores principales: Dharmashekar, Chandan, Shreevatsa, Bhargav, Jain, Anisha S., Harendra, Bhavana, Pradeep, Sushma, Vishwanath, Prashanth M., Singh, Pranav, V, Balamurugan, KK, Vinod, Patil, Sharanagouda S., Shati, Ali A., Alfaifi, Mohammad Y., Elbehairi, Serag Eldin I., Amachawadi, Raghavendra G., Kollur, Shiva Prasad, Shivamallu, Chandan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9920664/
https://www.ncbi.nlm.nih.gov/pubmed/36770771
http://dx.doi.org/10.3390/molecules28031106
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author Dharmashekar, Chandan
Shreevatsa, Bhargav
Jain, Anisha S.
Harendra, Bhavana
Pradeep, Sushma
Vishwanath, Prashanth M.
Singh, Pranav
V, Balamurugan
KK, Vinod
Patil, Sharanagouda S.
Shati, Ali A.
Alfaifi, Mohammad Y.
Elbehairi, Serag Eldin I.
Amachawadi, Raghavendra G.
Kollur, Shiva Prasad
Shivamallu, Chandan
author_facet Dharmashekar, Chandan
Shreevatsa, Bhargav
Jain, Anisha S.
Harendra, Bhavana
Pradeep, Sushma
Vishwanath, Prashanth M.
Singh, Pranav
V, Balamurugan
KK, Vinod
Patil, Sharanagouda S.
Shati, Ali A.
Alfaifi, Mohammad Y.
Elbehairi, Serag Eldin I.
Amachawadi, Raghavendra G.
Kollur, Shiva Prasad
Shivamallu, Chandan
author_sort Dharmashekar, Chandan
collection PubMed
description Bacterial infections are one of the leading causes of morbidity, mortality, and healthcare complications in patients. Leptospirosis is found to be the most prevalent, re-emergent, and neglected tropical zoonotic disease worldwide. The adaptation to various environmental conditions has made Leptospira acquire a large genome (~4.6 Mb) and a complex outer membrane, making it unique among bacteria that mimic the symptoms of jaundice and hemorrhage. Sph2 is another important virulence factor that enhances hemolytic sphingomyelinase—capable of moving inside mitochondria—which increases the ROS level and decreases the mitochondrial membrane potential, thereby leading to cell apoptosis. In the present study, 25 suspected bovine serum samples were subjected to the Microscopic Agglutination Test (MAT) across the Mysuru region. Different samples, such as urine, serum, and aborted materials from the confirmed MAT-positive animals, were used for isolation and genomic detection by conventional PCR targeting virulence gene, Lipl32, using specific primers. Further, in vitro and in silico studies were performed on isolated cultures to assess the anti-leptospiral, anti-hemolytic, and sphingomyelinase enzyme inhibition using novel pseudopeptides. The microdilution technique (MDT) and dark field microscope (DFM) assays revealed that at a concentration of 62.5 μg/mL, the pseudopeptide inhibited 100% of the growth of Leptospira spp., suggesting its efficiency in the treatment of leptospirosis. The flow cytometry analyses show the potency of the pseudopeptide against sphingomyelinase enzymes using human umbilical vein endothelial cells (HUVECs). Thus, the present study demonstrated the efficacy of the pseudopeptide in the inhibition of the growth of Leptospira, and therefore, this can be used as an alternative drug for the treatment of leptospirosis.
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spelling pubmed-99206642023-02-12 Evaluating the Antimicrobial and Anti-Hemolytic Activity of Synthesized Pseudopeptide against Leptospiral Species: In Silico and In Vitro Approach Dharmashekar, Chandan Shreevatsa, Bhargav Jain, Anisha S. Harendra, Bhavana Pradeep, Sushma Vishwanath, Prashanth M. Singh, Pranav V, Balamurugan KK, Vinod Patil, Sharanagouda S. Shati, Ali A. Alfaifi, Mohammad Y. Elbehairi, Serag Eldin I. Amachawadi, Raghavendra G. Kollur, Shiva Prasad Shivamallu, Chandan Molecules Article Bacterial infections are one of the leading causes of morbidity, mortality, and healthcare complications in patients. Leptospirosis is found to be the most prevalent, re-emergent, and neglected tropical zoonotic disease worldwide. The adaptation to various environmental conditions has made Leptospira acquire a large genome (~4.6 Mb) and a complex outer membrane, making it unique among bacteria that mimic the symptoms of jaundice and hemorrhage. Sph2 is another important virulence factor that enhances hemolytic sphingomyelinase—capable of moving inside mitochondria—which increases the ROS level and decreases the mitochondrial membrane potential, thereby leading to cell apoptosis. In the present study, 25 suspected bovine serum samples were subjected to the Microscopic Agglutination Test (MAT) across the Mysuru region. Different samples, such as urine, serum, and aborted materials from the confirmed MAT-positive animals, were used for isolation and genomic detection by conventional PCR targeting virulence gene, Lipl32, using specific primers. Further, in vitro and in silico studies were performed on isolated cultures to assess the anti-leptospiral, anti-hemolytic, and sphingomyelinase enzyme inhibition using novel pseudopeptides. The microdilution technique (MDT) and dark field microscope (DFM) assays revealed that at a concentration of 62.5 μg/mL, the pseudopeptide inhibited 100% of the growth of Leptospira spp., suggesting its efficiency in the treatment of leptospirosis. The flow cytometry analyses show the potency of the pseudopeptide against sphingomyelinase enzymes using human umbilical vein endothelial cells (HUVECs). Thus, the present study demonstrated the efficacy of the pseudopeptide in the inhibition of the growth of Leptospira, and therefore, this can be used as an alternative drug for the treatment of leptospirosis. MDPI 2023-01-22 /pmc/articles/PMC9920664/ /pubmed/36770771 http://dx.doi.org/10.3390/molecules28031106 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Dharmashekar, Chandan
Shreevatsa, Bhargav
Jain, Anisha S.
Harendra, Bhavana
Pradeep, Sushma
Vishwanath, Prashanth M.
Singh, Pranav
V, Balamurugan
KK, Vinod
Patil, Sharanagouda S.
Shati, Ali A.
Alfaifi, Mohammad Y.
Elbehairi, Serag Eldin I.
Amachawadi, Raghavendra G.
Kollur, Shiva Prasad
Shivamallu, Chandan
Evaluating the Antimicrobial and Anti-Hemolytic Activity of Synthesized Pseudopeptide against Leptospiral Species: In Silico and In Vitro Approach
title Evaluating the Antimicrobial and Anti-Hemolytic Activity of Synthesized Pseudopeptide against Leptospiral Species: In Silico and In Vitro Approach
title_full Evaluating the Antimicrobial and Anti-Hemolytic Activity of Synthesized Pseudopeptide against Leptospiral Species: In Silico and In Vitro Approach
title_fullStr Evaluating the Antimicrobial and Anti-Hemolytic Activity of Synthesized Pseudopeptide against Leptospiral Species: In Silico and In Vitro Approach
title_full_unstemmed Evaluating the Antimicrobial and Anti-Hemolytic Activity of Synthesized Pseudopeptide against Leptospiral Species: In Silico and In Vitro Approach
title_short Evaluating the Antimicrobial and Anti-Hemolytic Activity of Synthesized Pseudopeptide against Leptospiral Species: In Silico and In Vitro Approach
title_sort evaluating the antimicrobial and anti-hemolytic activity of synthesized pseudopeptide against leptospiral species: in silico and in vitro approach
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9920664/
https://www.ncbi.nlm.nih.gov/pubmed/36770771
http://dx.doi.org/10.3390/molecules28031106
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