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1.
Biochem Biophys Res Commun ; 695: 149452, 2024 02 05.
Artigo em Inglês | MEDLINE | ID: mdl-38169185

RESUMO

The osmotic pressure (Π) method has recently been developed to quantitatively examine the effect of membrane tension (σ) on pore formation in giant unilamellar vesicles (GUVs) induced by antimicrobial peptides (AMPs). Here, we used the Π method to reveal the effect of σ on the interaction of an AMP, PGLa, with lipid bilayers comprising dioleoylphosphatidylglycerol (DOPG) and dioleoylphosphatidylcholine (DOPC) (4/6). PGLa induced leakage of fluorescent probes from single GUVs under Π, indicating nanopore formation. Membrane tension did not transform a PGLa-induced nanopore into a micropore nor cause GUV burst up to 3.4 mN/m, which is in contrast with the effect of σ on another AMP, magainin 2-induced pore formation, where lower σ resulted in GUV burst. The fraction of leaking GUVs at a specific time increased with increasing σ, indicating that the rate of PGLa-induced pore formation increases with increasing σ. The rate of transfer of fluorescent probe-labeled PGLa across the lipid bilayer without pore formation also increased with increasing σ. PGLa-induced pore formation requires a symmetric distribution of peptides in both leaflets of the GUV bilayer, and thus we infer that the increase in the rate of PGLa transfer from the outer leaflet to the inner leaflet underlies the increase in the rate of pore formation with increasing σ. On the basis of these results, we discuss the difference between the effect of σ on nanopore formation in GUV membranes induced by PGLa and that by magainin 2.


Assuntos
Peptídeos Antimicrobianos , Bicamadas Lipídicas , Magaininas , Corantes Fluorescentes , Lipossomas Unilamelares
2.
Phys Chem Chem Phys ; 26(7): 6107-6117, 2024 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-38299672

RESUMO

The pore edge tension (Γ) of a membrane closely intertwines with membrane stability and plays a vital role in the mechanisms that facilitate membrane resealing following pore formation caused by electrical and mechanical tensions. We have explored a straightforward procedure to determine Γ by fitting the inverse of the tension-dependent logarithm of the rate constant of rupture of giant unilamellar vesicles (GUVs) using the Arrhenius equation. The GUVs were prepared using a combination of 1,2-dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DOPG) and 1, 2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) in a physiological environment. The effects of sugar concentration, membrane surface charge density, and membrane cholesterol concentration on Γ have been investigated. The values of Γ increase with sugar concentration in the physiological buffer, measuring 9.6 ± 0.3, 10.4 ± 0.1, and 16.2 ± 0.1 pN for 40, 100, and 300 mM, respectively. A higher concentration of anionic lipids (70 mol% of DOPG) significantly reduces Γ. An increasing trend of Γ with cholesterol content was observed; specifically, the values of Γ were 11.9 ± 0.9, 13.9 ± 0.7, and 16.2 ± 0.4 pN for 15, 29, and 40 mol% cholesterol, respectively. Thus, the presence of higher anionic lipids in the bilayer led to a decrease in membrane stability. In contrast, the presence of higher sugar concentrations in the buffer and increased cholesterol concentration in the membranes enhanced membrane stability.


Assuntos
Açúcares , Lipossomas Unilamelares , Fosfatidilcolinas , Cinética , Colesterol , Bicamadas Lipídicas
3.
J Chem Phys ; 160(1)2024 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-38165103

RESUMO

Positive membrane tension in the stretched plasma membrane of cells and in the stretched lipid bilayer of vesicles has been well analyzed quantitatively, whereas there is limited quantitative information on negative membrane tension in compressed plasma membranes and lipid bilayers. Here, we examined negative membrane tension quantitatively. First, we developed a theory to describe negative membrane tension by analyzing the free energy of lipid bilayers to obtain a theoretical equation for negative membrane tension. This allowed us to obtain an equation describing the negative membrane tension (σosm) for giant unilamellar vesicles (GUVs) in hypertonic solutions due to negative osmotic pressure (Π). Then, we experimentally estimated the negative membrane tension for GUVs in hypertonic solutions by measuring the rate constant (kr) of rupture of the GUVs induced by the constant tension (σex) due to an external force as a function of σex. We found that larger σex values were required to induce the rupture of GUVs under negative Π compared with GUVs in isotonic solution and quantitatively determined the negative membrane tension induced by Π (σosm) by the difference between these σex values. At small negative Π, the experimental values of negative σosm agree with their theoretical values within experimental error, but as negative Π increases, the deviation increases. Negative tension increased the stability of GUVs because higher tensions were required for GUV rupture, and the rate constant of antimicrobial peptide magainin 2-induced pore formation decreased.


Assuntos
Peptídeos Antimicrobianos , Bicamadas Lipídicas , Magaininas , Membrana Celular/metabolismo , Lipossomas Unilamelares , Soluções Hipertônicas
4.
Soft Matter ; 19(43): 8285-8304, 2023 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-37873600

RESUMO

This review provides an overview of the latest developments in both experimental and simulation techniques used to assess the bending rigidity of lipid membranes. It places special emphasis on experimental methods that utilize model vesicles to manipulate lipid compositions and other experimental parameters to determine the bending rigidity of the membrane. It also describes two commonly used simulation methods for estimating bending rigidity. The impact of various factors on membrane bending rigidity is summarized, including cholesterol, lipids, salt concentration, surface charge, membrane phase state, peptides, proteins, and polyethylene glycol. These factors are shown to influence the bending rigidity, contributing to a better understanding of the biophysical properties of membranes and their role in biological processes. Furthermore, the review discusses future directions and potential advancements in this research field, highlighting areas where further investigation is required.


Assuntos
Colesterol , Peptídeos , Membranas , Polietilenoglicóis , Bicamadas Lipídicas/química
5.
Phys Chem Chem Phys ; 24(11): 6716-6731, 2022 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-35234764

RESUMO

Osmotic pressure (Π) induces membrane tension in cells and lipid vesicles, which may affect the activity of antimicrobial peptides (AMPs) by an unknown mechanism. We recently quantitated the membrane tension of giant unilamellar vesicles (GUVs) due to Π under physiological conditions. Here, we applied this method to examine the effect of Π on the interaction of the AMP magainin 2 (Mag) with single GUVs. Under low Π values, Mag induced the formation of nanometer-scale pores, through which water-soluble fluorescent probe AF488 permeates across the membrane. The rate constant for Mag-induced pore formation (kp) increased with increasing Π. It has been proposed that the membrane tension in the GUV inner leaflet (σin) caused by Mag binding to the outer leaflet plays a vital role in Mag-induced pore formation. During the interactions between Mag and GUVs under Π, the σin increases due to Π, thereby increasing kp. The relationship between the kp and the total σin due to Π and Mag agreed with that without Π. In contrast, Mag induced rupture of a subset of GUVs under higher Π. Using fluorescence microscopy with a high-speed camera, the GUV rupture process was revealed. First, a small micrometer-scale pore was observed in individual GUVs. Then, the pore radius increased within ∼100 ms without changing the GUV diameter and concomitantly the thickness of the membrane at the pore rim increased, and finally the GUV transformed into a membrane aggregate. Based on these results, we discussed the effect of Π on Mag-induced damage of GUV membranes.


Assuntos
Peptídeos Antimicrobianos , Bicamadas Lipídicas , Magaininas , Pressão Osmótica , Lipossomas Unilamelares/metabolismo
7.
Biochim Biophys Acta Biomembr ; 1866(5): 184330, 2024 06.
Artigo em Inglês | MEDLINE | ID: mdl-38679311

RESUMO

To clarify the damage of lipid bilayer region in bacterial cell membrane caused by antimicrobial peptides (AMPs) and antimicrobial compounds (AMCs), their interactions with giant unilamellar vesicles (GUVs) of various lipid compositions have been examined. The findings revealed two main causes for the leakage: nanopore formation in the membrane and burst of GUVs. Although GUV burst has been explained previously based on the carpet model, the supporting evidence is limited. In this review, to better clarify the mechanism of GUV burst by AMPs, AMCs, and other membrane-active peptides, we described current knowledge of the conditions, characteristics, and detailed processes of GUV burst and the changes in the shape of the GUVs during burst. We identified several physical factors that affect GUV burst, such as membrane tension, electrostatic interaction, structural changes of GUV membrane such as membrane folding, and oil in the membrane. We also clarified one of the physical mechanisms underlying the instability of lipid bilayers that are associated with leakage in the carpet model. Based on these results, we propose a mechanism underlying some types of GUV burst induced by these substances: the growth of a nanopore to a micropore, resulting in GUV burst.


Assuntos
Peptídeos Antimicrobianos , Bicamadas Lipídicas , Lipossomas Unilamelares , Lipossomas Unilamelares/química , Lipossomas Unilamelares/metabolismo , Peptídeos Antimicrobianos/química , Peptídeos Antimicrobianos/farmacologia , Bicamadas Lipídicas/química , Bicamadas Lipídicas/metabolismo , Membrana Celular/química , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/farmacologia
8.
RSC Adv ; 14(36): 25986-26001, 2024 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-39161454

RESUMO

This review focuses on the deformation and poration of lipid vesicles caused by the interaction of anionic magnetite nanoparticles (MNPs). Effects of various factors, such as surface charge density, salt and sugar concentrations in buffer, membrane cholesterol content, polymer-grafted phospholipid, and membrane potential have been discussed for the interaction of MNPs with lipid vesicles. To quantify these effects on the vesicles, compactness, fraction of deformation and poration, dynamics of membrane permeation, and kinetics of membrane permeation have been critically evaluated. The review explores the potential advancements as well as future directions of the research field in the biomedical application of MNPs.

9.
J Phys Chem B ; 128(11): 2684-2696, 2024 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-38450565

RESUMO

Most antimicrobial peptides (AMPs) induce pore formation and a burst of lipid bilayers and plasma membranes. This causes severe leakage of the internal contents and cell death. The AMP PGLa forms nanopores in giant unilamellar vesicles (GUVs) comprising dioleoylphosphatidylcholine (DOPC) and dioleoylphosphatidylglycerol (DOPG). We here elucidated the effect of the line tension of a prepore rim on PGLa-induced nanopore formation by investigating the interaction of PGLa with single GUVs comprising dioleoylphosphatidylethanolamine (DOPE)/DOPG (6:4) in buffer using the single GUV method. We found that PGLa forms nanopores in the GUV membrane, which evolved into a local burst and burst of GUVs. The rate of pore formation in DOPE/DOPG-GUVs was smaller than that in DOPC/DOPG-GUVs. PGLa is located only in the outer leaflet of a GUV bilayer just before a fluorescent probe AF647 leakage from the inside, indicating that this asymmetric distribution induces nanopore formation. PGLa-induced local burst and burst of GUVs were observed at 10 ms-time resolution. After nanopore formation started, dense particles and small vesicles appeared in the GUVs, followed by a decrease in the GUV diameter. The GUV was finally converted into smaller GUV or lipid membrane aggregates. We discuss the mechanisms of PGLa-induced nanopore formation and its direct evolution to a GUV burst.


Assuntos
Peptídeos Antimicrobianos , Fosfatidiletanolaminas , Bicamadas Lipídicas/química , Lipossomas Unilamelares/química , Corantes Fluorescentes
10.
PLoS One ; 19(6): e0304345, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38857287

RESUMO

Irreversible electroporation induces permanent permeabilization of lipid membranes of vesicles, resulting in vesicle rupture upon the application of a pulsed electric field. Electrofusion is a phenomenon wherein neighboring vesicles can be induced to fuse by exposing them to a pulsed electric field. We focus how the frequency of direct current (DC) pulses of electric field impacts rupture and electrofusion in cell-sized giant unilamellar vesicles (GUVs) prepared in a physiological buffer. The average time, probability, and kinetics of rupture and electrofusion in GUVs have been explored at frequency 500, 800, 1050, and 1250 Hz. The average time of rupture of many 'single GUVs' decreases with the increase in frequency, whereas electrofusion shows the opposite trend. At 500 Hz, the rupture probability stands at 0.45 ± 0.02, while the electrofusion probability is 0.71 ± 0.01. However, at 1250 Hz, the rupture probability increases to 0.69 ± 0.03, whereas the electrofusion probability decreases to 0.46 ± 0.03. Furthermore, when considering kinetics, at 500 Hz, the rate constant of rupture is (0.8 ± 0.1)×10-2 s-1, and the rate constant of fusion is (2.4 ± 0.1)×10-2 s-1. In contrast, at 1250 Hz, the rate constant of rupture is (2.3 ± 0.8)×10-2 s-1, and the rate constant of electrofusion is (1.0 ± 0.1)×10-2 s-1. These results are discussed by considering the electrical model of the lipid bilayer and the energy barrier of a prepore.


Assuntos
Eletroporação , Lipossomas Unilamelares , Lipossomas Unilamelares/química , Cinética , Eletroporação/métodos , Probabilidade , Fusão de Membrana
11.
Sci Total Environ ; 926: 171944, 2024 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-38527542

RESUMO

Fluoroquinolone (FQ) antibiotics have become a subject of growing concern due to their increasing presence in the environment, particularly in the soil and groundwater. This review provides a comprehensive examination of the attributes, prevalence, ecotoxicity, and remediation approaches associated with FQs in environmental matrices. The paper discusses the physicochemical properties that influence the fate and transport of FQs in soil and groundwater, exploring the factors contributing to their prevalence in these environments. Furthermore, the ecotoxicological implications of FQ contamination in soil and aquatic ecosystems are reviewed, shedding light on the potential risks to environmental and human health. The latter part of the review is dedicated to an extensive analysis of remediation approaches, encompassing both in-situ and ex-situ methods employed to mitigate FQ contamination. The critical evaluation of these remediation strategies provides insights into their efficacy, limitations, and environmental implications. In this investigation, a correlation between FQ antibiotics and climate change is established, underlining its significance in addressing the Sustainable Development Goals (SDGs). The study further identifies and delineates multiple research gaps, proposing them as key areas for future investigational directions. Overall, this review aims to consolidate current knowledge on FQs in soil and groundwater, offering a valuable resource for researchers, policymakers, and practitioners engaged in environmental management and public health.


Assuntos
Antibacterianos , Ecossistema , Humanos , Antibacterianos/análise , Fluoroquinolonas/análise , Ecotoxicologia , Solo/química
12.
Biochim Biophys Acta Biomembr ; 1865(3): 184112, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-36567034

RESUMO

Most antimicrobial peptides (AMPs) damage the cell membrane of bacterial cells and induce rapid leakage of the internal cell contents, which is a main cause of their bactericidal activity. One of the AMPs, magainin 2 (Mag), forms nanopores in giant unilamellar vesicles (GUVs) comprising phosphatidylcholine (PC) and phosphatidylglycerol (PG), inducing leakage of fluorescent probes. In this study, to elucidate the Mag-induced pore formation in lipid bilayer region in E. coli cell membrane, we examined the interaction of Mag with single GUVs comprising E. coli polar lipids (E. coli-lipid-GUVs). First, we investigated the Mag-induced leakage of a fluorescent probe AF488 from single E. coli-lipid-GUVs, and found that Mag caused rupture of GUVs, inducing rapid AF488 leakage. The rate constant of Mag-induced GUV rupture increased with the Mag concentration. Using fluorescence microscopy with a time resolution of 5 ms, we revealed the GUV rupture process: first, a small micropore was observed in the GUV membrane, then the pore radius increased within 50 ms without changing the GUV diameter, the thickness of the membrane at the pore rim concomitantly increased, and eventually membrane aggregates were formed. Mag bound to only the outer monolayer of the GUV before GUV rupture, which increased the area of the GUV bilayer. We also examined the physical properties of E. coli-lipid-GUVs themselves. We found that the rate constant of the constant tension-induced rupture of E. coli-lipid-GUVs was higher than that of PG/PC-GUVs. Based on these results, we discussed the Mag-induced rupture of E. coli-lipid-GUVs and its mechanism.


Assuntos
Escherichia coli , Lipossomas Unilamelares , Magaininas/química , Lipossomas Unilamelares/química , Escherichia coli/metabolismo , Peptídeos Antimicrobianos , Bicamadas Lipídicas/química , Corantes Fluorescentes/metabolismo , Lecitinas/metabolismo
13.
Data Brief ; 51: 109741, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37965597

RESUMO

Speech Emotion Recognition (SER) identifies and categorizes emotional states by analyzing speech signals. SER is an emerging research area using machine learning and deep learning techniques due to its socio-cultural and business importance. An appropriate dataset is an important resource for SER related studies in a particular language. There is an apparent lack of SER datasets in Bangla language although it is one of the most spoken languages in the world. There are a few Bangla SER datasets but those consist of only a few dialogs with a minimal number of actors making them unsuitable for real-world applications. Moreover, the existing datasets do not consider the intensity level of emotions. The intensity of a specific emotional expression, such as anger or sadness, plays a crucial role in social behavior. Therefore, a realistic Bangla speech dataset is developed in this study which is called KUET Bangla Emotional Speech (KBES) dataset. The dataset consists of 900 audio signals (i.e., speech dialogs) from 35 actors (20 females and 15 males) with diverse age ranges. Source of the speech dialogs are Bangla Telefilm, Drama, TV Series, Web Series. There are five emotional categories: Neutral, Happy, Sad, Angry, and Disgust. Except Neutral, samples of a particular emotion are divided into two intensity levels: Low and High. The significant issue of the dataset is that the speech dialogs are almost unique with relatively large number of actors; whereas, existing datasets (such as SUBESCO and BanglaSER) contain samples with repeatedly spoken of a few pre-defined dialogs by a few actors/research volunteers in the laboratory environment. Finally, the KBES dataset is exposed as a nine-class problem to classify emotions into nine categories: Neutral, Happy (Low), Happy (High), Sad (Low), Sad (High), Angry (Low), Angry (High), Disgust (Low) and Disgust (High). However, the dataset is kept symmetrical containing 100 samples for each of the nine classes; 100 samples are also gender balanced with 50 samples for male/female actors. The developed dataset seems a realistic dataset while compared with the existing SER datasets.

14.
PLoS One ; 18(7): e0289087, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37523403

RESUMO

The hydrophilic polymer polyethylene glycol-grafted phospholipid has been used extensively in the study of artificial vesicles, nanomedicine, and antimicrobial peptides/proteins. In this research, the effects of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol)-2000] (abbreviated PEG-DOPE) on the deformation and poration of giant unilamellar vesicles (GUVs)-induced by anionic magnetite nanoparticles (NPs) have been investigated. For this, the size of the NPs used was 18 nm, and their concentration in the physiological solution was 2.00 µg/mL. GUVs were prepared using the natural swelling method comprising 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and PEG-DOPE. The mole% of PEG-DOPE in the membranes were 0, 2, and 5%. The degree of deformation of the GUVs was quantified by the parameter compactness (Com), which is 1.0 for the spherical-shaped GUVs. The value of Com increases with time during the interactions of NPs with GUVs for any concentration of PEG-DOPE, but the rate of increase is significantly influenced by the PEG-DOPE concentration in the membranes. The average compactness increases with the increase of PEG-DOPE%, and after 60 min of NPs interaction, the values of average compactness for 0, 2, and 5% PEG-DOPE were 1.19 ± 0.02, 1.26 ± 0.03 and 1.35 ± 0.05, respectively. The fraction of deformation (Frd) also increased with the increase of PEG-DOPE%, and at 60 min, the values of Frd for 0 and 5% PEG-DOPE were 0.47 ± 0.02 and 0.63 ± 0.02, respectively. The fraction of poration (Frp) increased with the increase of PEG-DOPE, and at 60 min, the values of Frp for 0 and 5% PEG-DOPE were 0.25 ± 0.02 and 0.48 ± 0.02, respectively. Hence, the presence of PEG-grafted phospholipid in the membranes greatly enhances the anionic magnetite NPs-induced deformation and poration of giant vesicles.


Assuntos
Nanopartículas de Magnetita , Fosfolipídeos , Polietilenoglicóis , Polímeros , Lipossomas Unilamelares , Fosfatidilcolinas
15.
Sci Total Environ ; 876: 162414, 2023 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-36868275

RESUMO

The occurrence of microplastics (MPs) in aquatic environments has been a global concern because they are toxic and persistent and may serve as a vector for many legacies and emerging pollutants. MPs are discharged to aquatic environments from different sources, especially from wastewater plants (WWPs), causing severe impacts on aquatic organisms. This study mainly aims to review the Toxicity of MPs along with plastic additives in aquatic organisms at various trophic compartments and available remediation methods/strategies for MPs in aquatic environments. Occurrences of oxidative stress, neurotoxicity, and alterations in enzyme activity, growth, and feeding performance were identical in fish due to MPs toxicity. On the other hand, growth inhibition and ROS formation were observed in most of the microalgae species. In zooplankton, potential impacts were acceleration of premature molting, growth retardation, mortality increase, feeding behaviour, lipid accumulation, and decreased reproduction activity. MPs togather with additive contaminants could also exert some toxicological impacts on polychaete, including neurotoxicity, destabilization of the cytoskeleton, reduced feeding rate, growth, survivability and burrowing ability, weight loss, and high rate of mRNA transcription. Among different chemical and biological treatments for MPs, high removal rates have been reported for coagulation and filtration (>86.5 %), electrocoagulation (>90 %), advanced oxidation process (AOPs) (30 % to 95 %), primary sedimentation/Grit chamber (16.5 % to 58.84 %), adsorption removal technique (>95 %), magnetic filtration (78 % to 93 %), oil film extraction (>95 %), and density separation (95 % to 100 %). However, desirable extraction methods are required for large-scale research in MPs removal from aquatic environments.


Assuntos
Microplásticos , Poluentes Químicos da Água , Animais , Plásticos/toxicidade , Poluentes Químicos da Água/análise , Águas Residuárias , Peixes , Organismos Aquáticos
16.
Environ Sci Pollut Res Int ; 30(45): 100828-100844, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37644270

RESUMO

Tobacco products are widely recognized as a major contributor to death. Cigarette smoke contains several toxic chemicals including heavy metals particulate causing high health risks. However, limited information has been available on the health risks associated with the heavy metals in cigarettes commonly sold in the Bangladeshi market. This study evaluated the concentrations and potential health risks posed by ten concerned heavy metals in ten widely consumed cigarette brands in Bangladesh using an atomic absorption spectrometer. The concentration (mg/kg) ranges of heavy metals Pb, Cd, Cr, As, Co, Ni, Mn, Fe, Cu, and Zn vary between 0.46-1.05, 0.55-1.03, 0.80-1.2, 0.22-0.40, 0.46-0.78, 2.59-3.03, 436.8-762.7, 115.8-184.4, 146.6-217.7, and 34.0-42.7, respectively. We assume that the heavy metals content among cigarette brands is varied due to the differences in the source of tobacco they use for cigarette preparation. The carcinogenic risks posed by heavy metals follow the order of Cr > Co > Cd > As > Ni > Pb, while the non-carcinogenic risks for Cu, Zn, Fe, and Mn were greater than unity (HQ > 1), except for Fe. The existence of toxic heavy metals in cigarette tobacco may thus introduce noticeable non-carcinogenic and carcinogenic health impacts accompanying inhalation exposure. This study provides the first comprehensive report so far on heavy metal concentration and associated health risks in branded cigarettes commonly sold in Bangladesh. Hence, this data and the information provided can serve as a baseline as well as a reference for future research and have potential implications for policy and legislation in Bangladesh.

17.
Microbiol Spectr ; 10(4): e0011422, 2022 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-35863040

RESUMO

Antimicrobial peptides (AMPs) inhibit the proliferation of or kill bacterial cells. To measure these activities, several methods have been used, which provide only the average value of many cells. Here, we report the development of a method to examine the antimicrobial and bactericidal activities of AMPs at the single-cell level (i.e., single-cell analysis) and apply this strategy to examine the interaction of an AMP, magainin 2 (Mag), with Escherichia coli cells. Using this method, we monitored the proliferation of single cells on agar in a microchamber and measured the distribution of the number of cells in each microcolony using optical microscopy. For method A, we incubated cells in the presence of various concentrations of AMPs for 3 h. The fraction of microcolonies containing only a single cell, Psingle, increased with the Mag concentration and reached 1 at a specific concentration, which corresponded to the MIC. For method B, after the interaction of a cell suspension with an AMP for a specific time, an aliquot was diluted to stop the interaction, and the proliferation of single cells then was monitored after a 3-h incubation; this method permits the definition of Psingle(t), the fraction of dead cells after the interaction. For the interaction of Mag with E. coli cells, Psingle(t) increased with the interaction time, reaching ~1 at 10 and 20 min for 25 and 13 µM Mag, respectively. Thus, these results indicate that a short interaction time between Mag and E. coli cells is sufficient to induce bacterial cell death. IMPORTANCE To elucidate the activity of antimicrobial peptides (AMPs) against bacterial cells, it is important to estimate the interaction time that is sufficient to induce cell death. We have developed a method to examine the antimicrobial and bactericidal activities of AMPs at the single-cell level (i.e., single-cell analysis). Using this method, we monitored the proliferation of single cells on agar in a microchamber and measured the distribution of the number of cells in each microcolony using optical microscopy. We found that during the interaction of magainin 2 (Mag) with E. coli cells, the fraction of dead cells, Psingle(t), increased with the interaction time, rapidly reaching 1 (e.g., 10 min for 25 µM Mag). This result indicates that Mag induces cell death after a short time of interaction.


Assuntos
Anti-Infecciosos , Peptídeos Antimicrobianos , Monofosfato de Adenosina , Ágar , Antibacterianos/farmacologia , Anti-Infecciosos/farmacologia , Escherichia coli , Magaininas/química , Magaininas/farmacologia , Testes de Sensibilidade Microbiana , Análise de Célula Única
18.
Environ Pollut ; 293: 118549, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34813884

RESUMO

Ocean acidification (OA) has received more attention in the marine research community in recent years than any other topic. Excess carbon dioxide makes the ocean more acidic, threatening marine ecosystems. There has been little research on the impact of OA on crustaceans, particularly on their physiological and potential ecosystem-level consequences. Thus, we investigated the impacts of OA on the physiological and biochemical characteristics of the estuarine amphipod Ampelisca brevicornis. Ovigerous amphipods were harvested from nature and maintained in the laboratory to produce juveniles, which were then further reared to obtain the mature adults (F0) and successive offspring (F1). For this study, four pH treatments (pH 8.1, 7.5, 7.0, and 6.5) mimicking future OA were evaluated to understand the physiological and biochemical effects on the organisms. The findings of this study suggest that A. brevicornis is more vulnerable to OA than was previously established in short-term trials. The survival was significantly reduced as pH decreased over time and a significant interaction between pH and time was observed. Survival was higher in F1 than in F0 juveniles and vice versa in terms of growth. Animal's physiological responses such as growth, burrowing behavior, locomotor activity, swimming speed, ventilation rate and reproductive performance were negatively influenced by acidification. These physiological characteristics can be linked to the oxidative stress induced by global change conditions because excess of free radicals degrade cell functioning, affecting species' biochemical and physiological performance. These alterations may have long-term negative impacts, with ecological consequences. The results of this study provide baseline information regarding the effect of OA on this keystone crustacean that may be useful in simulating the impacts of OA to develop different conceptual models for a better understanding of the consequences and implications of climate change in the future for managing marine ecosystems.


Assuntos
Anfípodes , Animais , Ecossistema , Concentração de Íons de Hidrogênio , Oceanos e Mares , Água do Mar
19.
Environ Sci Pollut Res Int ; 29(11): 15347-15363, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34989993

RESUMO

Coastal wetlands including salt marshes are among the most productive ecosystems on Earth. They are known for improving the quality of coastal water and provisioning coastal fisheries. However, this ecosystem is under potential threat due to urban coastal land reclamation, limited sediment supply, increased nutrient/eutrophication, and sea level rise. Therefore, restoration efforts to protect the degraded salt marsh habitat are considerably increasing worldwide. In this paper, we present an overview of salt marsh restoration techniques and success indicators. Published scientific literature in English language was collected by searching the most relevant keywords from popular search engines, namely, Google Scholar, Scopus, and Mendeley to get the information about salt marsh restoration techniques and success indicators. This study comprehensively reviewed data from 78 peer-reviewed papers. Results indicated that much of the salt marsh was restored through assisted abiotic strategies (e.g., recovery of tidal exchange, managed realignment, and sediment level amendment). A total of 214 indicators were found, spanning over six major ecological attributes such as structural diversity, ecosystem functions, physical conditions, species composition, external exchange, and absence of threat. Author keywords analysis revealed several hotspots for recent research (e.g., 16 s rRNA, fungi, microbial communities, carbon accumulation, and blue carbon). This paper proposes a model for restoring degraded salt marsh, as well as tracking their success. The information presented here will assist the marine ecosystem restoration practitioners in getting a comprehensive understanding of salt marsh restoration success evaluation.


Assuntos
Ecossistema , Áreas Alagadas , Carbono , Eutrofização
20.
Trop Med Health ; 50(1): 11, 2022 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-35078540

RESUMO

Dengue fever is a viral infection caused by Aedes mosquitoes that has recently expanded fast in many of the WHO member states globally. Female mosquitoes, mostly Aedes aegypti and, to a smaller degree, Aedes albopictus, disseminate dengue virus. Dengue fever has been more common in recent decades all across the world, and Bangladesh is no exception. As the COVID-19 outbreak wreaks havoc, the following rise in dengue illnesses has been a source of considerable concern. As the health care has been stretched thin in these dangerous times, the vulnerable population has been left at the mercy of these two viral infections. Lack of knowledge, major legislative changes, poor eradication initiatives, and a lack of financing resources have all contributed to the increase in numbers. Stakeholders and policymakers must begin taking meaningful actions and implementing well-thought-out adjustments immediately, or the situation will worsen, resulting in the loss of thousands of innocent lives.

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