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1.
Heliyon ; 10(9): e29506, 2024 May 15.
Article in English | MEDLINE | ID: mdl-38698983

ABSTRACT

Public transportation plays a critical role in meeting transportation demands, particularly in densely populated areas. The COVID-19 pandemic has highlighted the importance of public health measures, including the need to prevent the spread of the virus through public transport. The spreading of the virus on a passenger ship is studied using the Computational Fluid Dynamic (CFD) model and Monte Carlo simulation. A particular focus was the context of Bangladesh, a populous maritime nation in South Asia, where a significant proportion of the population utilizes passenger ships to meet transportation demands. In this regard, a turbulence model is used, which simulates the airflow pattern and determines the contamination zone. Parameters under investigation are voyage duration, number of passengers on board, social distance, the effect of surgical masks, and others. This study shows that the transmission rate of SARS-CoV-2 infection on public transport, such as passenger ships, is not necessarily directly proportional to voyage duration or the number of passengers onboard. This model has the potential to be applied in various other modes of transportation, including public buses and airplanes. Implementing this model may help to monitor and address potential health risks effectively in the public transport networks.

2.
J Health Popul Nutr ; 43(1): 64, 2024 May 13.
Article in English | MEDLINE | ID: mdl-38741173

ABSTRACT

BACKGROUND: The high incidence of low birth weight (LBW) is associated with an increased risk of infant mortality, adverse pregnancy outcomes for mothers, and a decline in overall health and well-being. The current study aimed to identify the various determinants of LBW and its effect on adverse health and nutritional outcomes of children aged 0-23 months in Bangladesh. METHODS: Bangladesh Demography and Health Survey (BDHS) 2017-18 data was used. A chi-square test and multivariable logistic regression analysis were used to find out the associations between independent variables and outcomes (e.g., LBW, child illness and undernutrition). RESULTS: The overall prevalence of LBW among was 16.3%. Mother with no formal education (AOR = 2.64, 95% CI = 0.55-3.30, p = 0.01), female child (AOR = 1.31, 95% CI = 1.04-1.65, p = 0.023); and poorest economic status (AOR = 1.69, 95% CI = 1.13-2.51, p = 0.010), were identified significant determinants of LBW. Of home environment and hygiene factors, unimproved toilet facilities (AOR = 1.38, 95% CI = 1.03-1.84, p = 0.030) had a significant effect on LBW. In addition, children born with LBW were more likely to suffer fever (AOR = 1.26, 95% CI = 1.05-1.60, p = 0.050), stunting (AOR = 2.42, 95% CI = 1.86-3.15, p = < 0.001), wasting (AOR = 1.47, 95% CI = 1.02-2.25 p = 0.049), and underweight (AOR = 3.19, 95% CI = 2.40-4.23, p = < 0.001). CONCLUSION: One out of five children was LBW in Bangladesh. Maternal education, sex of child, wealth index, and toilet facilities had significant effects on LBW. In addition, LWB contributed to children's poor health and nutritional outcomes. Enhancing maternal pregnancy, and child health outcomes necessitates policies addressing poverty, gender inequality, and social disparities. Key strategies include promoting regular prenatal care, early medical intervention, reproductive health education, and safe hygiene practices. To combat the negative impacts of LBW, a comprehensive strategy is vital, encompassing exclusive breastfeeding, nutritional support, growth monitoring, accessible healthcare, and caregiver education.


Subject(s)
Child Health , Infant, Low Birth Weight , Humans , Bangladesh/epidemiology , Female , Infant, Newborn , Infant , Male , Adult , Child Health/statistics & numerical data , Young Adult , Prevalence , Nutritional Status , Risk Factors , Socioeconomic Factors , Health Surveys , Pregnancy , Adolescent , Cross-Sectional Studies
3.
Plants (Basel) ; 13(5)2024 Feb 20.
Article in English | MEDLINE | ID: mdl-38475410

ABSTRACT

The octoploid-cultivated strawberry variety Benihope (Fragaria × ananassa Duch cv. Benihope) is an important commercial plant. It is highly susceptible to different diseases, which ultimately leads to a reduction in yield. Gene-editing methods, such as CRISPR/Cas9, demonstrate potential for improving disease resistance in the strawberry cv. Benihope. Establishing a plant regeneration system suitable for CRISPR/Cas9 gene editing is crucial for obtaining transgenic plants on a large scale. This research established a callus induction and plant regeneration system for Agrobacterium-mediated CRISPR/Cas9 gene editing in strawberry cv. Benihope by evaluating multiple types of explants and various plant growth regulators throughout the entire tissue culture process. The results showed that the efficiency of callus induction is strongly influenced by the type of explant and is highly sensitive to the combination of plant growth regulators. Among the different plant growth regulators employed, thidiazuron (TDZ), in combination with 2,4-dichlorophenoxyacetic acid (2,4-D), effectively induced callus formation and plant regeneration from explants derived from nutrient tissues such as runner tips and crowns. In addition, the regeneration experiment demonstrated that the addition of polyvinylpyrrolidone (PVPP) to the shoot regeneration medium could inhibit tissue browning. The gene-edited plants in which some or all of the Fvb7-1, Fvb7-2, Fvb7-3, and Fvb7-4 genes in the MLO (Mildew resistance Locus O) gene family were knocked out by CRISPR/Cas9 system were obtained by applying the plant regeneration system developed in this study.

4.
J Biomed Mater Res A ; 2024 Mar 15.
Article in English | MEDLINE | ID: mdl-38487991

ABSTRACT

Mesenchymal stem cell-derived secretome represents an emerging acellular therapeutic which possess significant opportunity for clinical applications due to its anti-inflammatory, immunomodulatory, and wound healing properties. However, maintaining therapeutic efficacy and ensuring stability of cell-based products is challenging, requiring a robust delivery method. Therefore, we designed a hydrogel-based scaffold loaded with CK Cell Technologies' proprietary Mesenchymal stem cell-secretome for controlled release treatment of acute and chronic wounds. We incorporated both conditioned media (CM) and extracellular vesicles (EVs) into gelatin methacryloyl (GelMA) hydrogels and demonstrated how we can tune the diffusive release of the EVs from them. To demonstrate viability of the approach, we developed a wound healing scratch assay where we see in situ release of CM and EVs promote enhanced migration of human dermal fibroblasts (hDFs). We see the colocalization of these EVs in the fibroblasts using fluorescent microscopy. Finally, as a surrogate for in vivo neovascularization, we conducted an in vitro tube formation assay for the MSC-secretome using matrigel-embedded human microvascular endothelial cells. By adding CM and EVs, we observe an increase in tubulogenesis. Collectively, our data demonstrates by tuning the GelMA properties, we can influence the controlled release of the MSC-secretome for a wound dressing and bandage application for chronic and acute wounds.

5.
Heliyon ; 10(3): e25278, 2024 Feb 15.
Article in English | MEDLINE | ID: mdl-38317892

ABSTRACT

The utilization of synthetic dyes in food industries is a great concern for food safety and health issues. So, natural pigments can be an excellent substitute for synthetic dyes and also health-friendly for consumers. In the experiment, natural pigments were extracted from tomato (Solanum lycopersicum L.), turmeric (Curcuma longa L.) and red amaranth (Amaranthus tricolor L.). Then the stability and consumer acceptance of the extracted pigments were examined. The highest amount of pigment was extracted from turmeric (2.14 ± 0.30 %) with ethanol solvent, followed by tomato (0.67 ± 0.06 %) with hexane: acetone (1:1) solvent, and red amaranth (0.78 ± 0.05 %) with acetone solvent. Turmeric pigment showed the highest stability in high temperatures and light exposure. All of the pigments were highly stable in a neutral environment; however, tomato pigment showed the highest stability index (84.33 ± 2.52) at pH 3.0, but turmeric pigment showed the highest stability (91.67 ± 1.53) at pH 5.0. The simple preference test revealed that the use of turmeric pigment in boiled rice had the highest acceptance rate, and in terms of taste and flavor, red amaranth pigments in ice cream. So turmeric pigment can be utilized in high-temperature processing and/or acidic foods, but tomato and red amaranth pigments might be in low-temperature processing foods such as the ice-cream and soft drinks processing industry.

6.
J Phys Chem B ; 128(2): 429-439, 2024 Jan 18.
Article in English | MEDLINE | ID: mdl-38179652

ABSTRACT

The KCNA5 gene provides the code for the α-subunit of the potassium channel Kv1.5. The genetic variant H463R in the Kv1.5 channel has been reported to cause a functional loss in atrial fibrillation (AF) patients. Understanding the mutations at a molecular level is key to developing improved therapeutics concerning cardiac hKv1.5 and hKv1.4 channels. Molecular dynamics and umbrella sampling free energy simulations are an effective tool to understand the mutation's effect on ion conduction, which we have employed and found that the hKv1.5[H463R] mutation imposes an energy barrier on the ion conduction pathway compared to the wild-type channel's ion free energy and pore structure. These results imply that the arginine mutation associated with the AF disease in particular modulates the inactivation process of hKv1.5. Kv1.4, encoded by the KCNA4 gene, is also present in the heart. Therefore, we considered simulation studies of the equivalent H507R mutation in the hKv1.4 channel and found that the mutation slightly reduces the ion conduction barrier in the ion conduction pathway, making it insignificant.


Subject(s)
Molecular Dynamics Simulation , Potassium Channels , Humans , Mutation , Potassium Channels/metabolism , Heart , Potassium Channel Blockers/chemistry , Potassium Channel Blockers/metabolism
7.
Curr Diabetes Rev ; 20(3): e150523216913, 2024.
Article in English | MEDLINE | ID: mdl-37190800

ABSTRACT

The human body is a complex ecosystem that thrives on symbiosis. It is estimated that around 10^14 commensal microorganisms inhabit the human body, with the gut microbiota being one of the most diverse and complex populations of bacteria. This community is thought to comprise over a thousand different species that play a crucial role in the development of critical human diseases such as cancer, obesity, diabetes, mental depression, hypertension, and others. The gut microbiota has been identified as one of the most recent contributors to these metabolic disorders. With the emergence of inexpensive and high-performance sequence technology, our understanding of the function of the intestinal microbiome in host metabolism regulation and the development of (cardio) metabolic diseases has increased significantly. The symbiotic relationship between the gut microbiota and the host is essential for properly developing the human metabolic system. However, if this balance is disrupted by various factors such as infection, diet, exercise, sleep patterns, or exposure to antibiotics, it can lead to the development of various diseases in the body, including obesity and diabetes type 1 and 2. While many approaches and medications have been developed globally to treat these diseases, none have proven to be entirely effective, and many show side effects. Therefore, scientists believe that treating the gut microbiota using tried-and-true methods is the best option for combating obesity and diabetes. In this study, we aim to identify several feasible ways and prospects for gut microbiota therapy that can shape a new format for the treatment of obesity and diabetes.


Subject(s)
Diabetes Mellitus , Gastrointestinal Microbiome , Humans , Diet , Gastrointestinal Microbiome/physiology , Obesity/metabolism
8.
ACS Omega ; 8(48): 45700-45707, 2023 Dec 05.
Article in English | MEDLINE | ID: mdl-38075789

ABSTRACT

This study showcases the creation of a biosensor strip designed for the rapid, precise, and highly sensitive electrochemical detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). These biosensor strips were crafted by affixing a monoclonal antibody (mAb) specific to SARS-CoV-2 onto the surface of a commercially screen-printed carbon electrode (SPCE) modified with polyaniline-titania nanotubes (PANi-TNT). The transportable sensing device was constructed by pairing the mAb functionalized strip with a portable potentiostat wirelessly connected to either a Windows or Android device. Fast and specific conjugation between spike protein of SARS-CoV-2 and immobilized anti-SARS-CoV-2 triggered a change in the charge and electron mobility in the biosensing layer of the strip to produce detectable current during chronoamperometric scanning in the presence of a phosphate buffer solution (PBS). The excellent sensitivity and specificity of the sensor toward SARS-CoV-2 were detected as analytical analysis demonstrated linearity in the range of 80 to 200 copies/µL with a limit of detection of 25.59 copies/µL from the dose-response and standard fitted curve. Through experimental validation, the sensor strip's ability to specifically detect SARS-CoV-2 was established, distinguishing it from human coronavirus-OC43 (HCoV-OC43), HCoV-NL63, HCoV-229E, and adenovirus. The results from these tests indicate that these strips possess the potential for the future creation of dependable and easily transportable point-of-care diagnostic devices, enabling swift, sensitive, and precise detection of SARS-CoV-2 in the saliva or nasopharyngeal fluid of individuals infected with the virus.

9.
Children (Basel) ; 10(11)2023 Nov 20.
Article in English | MEDLINE | ID: mdl-38002920

ABSTRACT

Diarrhea and fever are prevalent childhood illnesses with potentially severe consequences, especially when they co-occur. This study investigates the prevalence and determinants of diarrhea, fever, and their coexistence among children under-five in Bangladesh. Data from the 2017-2018 Bangladesh Demography and Health Survey (BDHS) were analyzed using multivariable stepwise logistic regression with backward selection. This study found that 5.0% for diarrhea, 34.0% for fever, and 3.0% for the coexistence of both illnesses. Common factors associated with childhood diarrhea and fever included the child's age (12-23 months), and the mother's education. Diarrhea was associated with households with improved water sources and children in the Barisal division, while fever was linked to underweight children and those from more affluent backgrounds. The coexistence of both was significantly linked to underweight children, higher birth orders, and children from the Rajshahi division. Notably, child illnesses were associated with parental education, higher socio-economic status, and access to improved drinking water sources. Diarrhea affects one in 20 children, fever affects one in three, and the coexistence of both conditions affects one in 35 children in Bangladesh. The findings need further research and policy reviews to develop effective interventions and improve child health in Bangladesh.

10.
Mol Cell Proteomics ; 22(12): 100679, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37979947

ABSTRACT

The ability of an organism to respond to environmental changes is paramount to survival across a range of conditions. The bacterial heme nitric oxide/oxygen binding proteins (H-NOX) are a family of biofilm-regulating gas sensors that enable bacteria to respond accordingly to the cytotoxic molecule nitric oxide. By interacting with downstream signaling partners, H-NOX regulates the production of the bacterial secondary messenger cyclic diguanylate monophosphate (c-di-GMP) to influence biofilm formation. The aquatic organism Caulobacter crescentus has the propensity to attach to surfaces as part of its transition into the stalked S-phase of its life cycle. This behavior is heavily influenced by intracellular c-di-GMP and thus poses H-NOX as a potential influencer of C. crescentus surface attachment and cell cycle. By generating a strain of C. crescentus lacking hnox, our laboratory has demonstrated that this strain exhibits a considerable growth deficit, an increase in biofilm formation, and an elevation in c-di-GMP. Furthermore, in our comprehensive proteome study of 2779 proteins, 236 proteins were identified that exhibited differential expression in Δhnox C. crescentus, with 132 being downregulated and 104 being upregulated, as determined by a fold change of ≥1.5 or ≤0.66 and a p value ≤0.05. Our systematic analysis unveiled several regulated candidates including GcrA, PopA, RsaA, FtsL, DipM, FlgC, and CpaE that are associated with the regulation of the cellular division process, surface proteins, flagellum, and pili assembly. Further examination of Gene Ontology and pathways indicated that the key differences could be attributed to several metabolic processes. Taken together, our data indicate a role for the HNOX protein in C. crescentus cell cycle progression.


Subject(s)
Caulobacter crescentus , Hemeproteins , Caulobacter crescentus/genetics , Caulobacter crescentus/metabolism , Nitric Oxide/metabolism , Cyclic GMP/metabolism , Hemeproteins/genetics , Hemeproteins/metabolism , Oxygen/metabolism , Bacterial Proteins/metabolism , Cell Cycle , Heme/metabolism , Gene Expression Regulation, Bacterial
11.
J Evid Based Med ; 16(3): 303-320, 2023 Sep.
Article in English | MEDLINE | ID: mdl-37691394

ABSTRACT

AIM: Technology including artificial intelligence (AI) may play a key role to strengthen primary health care services in resource-poor settings. This systematic review aims to explore the evidence on the use of AI and digital health in improving primary health care service delivery. METHODS: Three electronic databases were searched using a comprehensive search strategy without providing any restriction in June 2023. Retrieved articles were screened independently using the "Rayyan" software. Data extraction and quality assessment were conducted independently by two review authors. A narrative synthesis of the included interventions was conducted. RESULTS: A total of 4596 articles were screened, and finally, 48 articles were included from 21 different countries published between 2013 and 2021. The main focus of the included studies was noncommunicable diseases (n = 15), maternal and child health care (n = 11), primary care (n = 8), infectious diseases including tuberculosis, leprosy, and HIV (n = 7), and mental health (n = 6). Included studies considered interventions using AI, and digital health of which mobile-phone-based interventions were prominent. m-health interventions were well adopted and easy to use and improved the record-keeping, service deliver, and patient satisfaction. CONCLUSION: AI and the application of digital technologies improve primary health care service delivery in resource-poor settings in various ways. However, in most of the cases, the application of AI and digital health is implemented through m-health. There is a great scope to conduct further research exploring the interventions on a large scale.

12.
Article in English | MEDLINE | ID: mdl-37643902

ABSTRACT

The extracellular matrix in tissue consists of complex heterogeneous soft materials with hierarchical structure and dynamic mechanical properties dictating cell and tissue level function. In many natural matrices, there are nanofibrous structures that serve to guide cell activity and dictate the form and function of tissue. Synthetic hydrogels with integrated nanofibers can mimic the structural properties of native tissue; however, model systems with dynamic mechanical properties remain elusive. Here we demonstrate modular nanofibrous hydrogels that can be reversibly stiffened in response to applied magnetic fields. Iron oxide nanoparticles were incorporated into gelatin nanofibers through electrospinning, followed by chemical stabilization and fragmentation. These magnetoactive nanofibers can be mixed with virtually any hydrogel material and reversibly stiffen the matrix at a low fiber content (≤3%). In contrast to previous work, where a large quantity of magnetic material disallowed cell encapsulation, the low nanofiber content allows matrix stiffening with cells in 3D. Using adipose derived stem cells, we show how nanofibrous matrices are beneficial for both osteogenesis and adipogenesis, where stiffening the hydrogel with applied magnetic fields enhances osteogenesis while discouraging adipogenesis. Skeletal myoblast progenitors were used as a model of tissue morphogenesis with matrix stiffening augmenting myogenesis and multinucleated myotube formation. The ability to reversibly stiffen fibrous hydrogels through magnetic stimulation provides a useful tool for studying nanotopography and dynamic mechanics in cell culture, with a scope for stimuli responsive materials for tissue engineering.

15.
Hum Genomics ; 17(1): 58, 2023 Jul 07.
Article in English | MEDLINE | ID: mdl-37420264

ABSTRACT

BACKGROUND: Wastewater-based epidemiological surveillance has been considered a powerful tool for early detection and monitoring of the dynamics of SARS-CoV-2 and its lineages circulating in a community. This study is aimed to investigate the complexity of SARS-CoV-2 infection dynamics in Dhaka city by examining its genetic variants in wastewater. Also, the study seeks to determine a connection between the SARS-CoV-2 variations detected in clinical testing and those found in wastewater samples. RESULTS: Out of 504 samples tested in RT-qPCR, 185 (36.7%) tested positive for SARS-CoV-2 viral RNA. The median log10 concentration of SARS-CoV-2 N gene copies/Liter of wastewater (gc/L) was 5.2, and the median log10 concentration of ORF1ab was 4.9. To further reveal the genetic diversity of SARS-CoV-2, ten samples with ORF1ab real-time RT-PCR cycle threshold (Ct) values ranging from 28.78 to 32.13 were subjected to whole genome sequencing using nanopore technology. According to clade classification, sequences from wastewater samples were grouped into 4 clades: 20A, 20B, 21A, 21J, and the Pango lineage, B.1, B.1.1, B.1.1.25, and B.1.617.2, with coverage ranging from 94.2 to 99.8%. Of them, 70% belonged to clade 20B, followed by 10% to clade 20A, 21A, and 21J. Lineage B.1.1.25 was predominant in Bangladesh and phylogenetically related to the sequences from India, the USA, Canada, the UK, and Italy. The Delta variant (B.1.617.2) was first identified in clinical samples at the beginning of May 2021. In contrast, we found that it was circulating in the community and was detected in wastewater in September 2020. CONCLUSION: Environmental surveillance is useful for monitoring temporal and spatial trends of existing and emerging infectious diseases and supports evidence-based public health measures. The findings of this study supported the use of wastewater-based epidemiology and provided the baseline data for the dynamics of SARS-CoV-2 variants in the wastewater environment in Dhaka, Bangladesh.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , SARS-CoV-2/genetics , Bangladesh/epidemiology , COVID-19/epidemiology , Public Health Surveillance , Wastewater , Complement System Proteins , RNA
17.
Environ Sci Pollut Res Int ; 30(29): 73393-73404, 2023 Jun.
Article in English | MEDLINE | ID: mdl-37188935

ABSTRACT

In the present study, we determined the developmental toxicity of endosulfan at an elevated ambient temperature using the zebrafish animal model. Zebrafish embryos of various developmental stages were exposed to endosulfan through E3 medium, raised under two selected temperature conditions (28.5 °C and an elevated temperature of 35 °C), and monitored under the microscope. Zebrafish embryos of very early developmental stages (cellular cleavage stages, such as the 64-cell stage) were highly sensitive to the elevated temperature as 37.5% died and 47.5% developed into amorphous type, while only 15.0% of embryos developed as normal embryos without malformation. Zebrafish embryos that were exposed concurrently to endosulfan and an elevated temperature showed stronger developmental defects (arrested epiboly progress, shortened body length, curved trunk) compared to the embryos exposed to either endosulfan or an elevated temperature. The brain structure of the embryos that concurrently were exposed to the elevated temperature and endosulfan was either incompletely developed or malformed. Furthermore, the stress-implicated genes hsp70, p16, and smp30 regulations were synergistically affected by endosulfan treatment under the elevated thermal condition. Overall, the elevated ambient temperature synergistically enhanced the developmental toxicity of endosulfan in zebrafish embryos.


Subject(s)
Endosulfan , Zebrafish , Animals , Endosulfan/toxicity , Temperature , Embryonic Development , Embryo, Nonmammalian/abnormalities
18.
Heliyon ; 9(5): e15709, 2023 May.
Article in English | MEDLINE | ID: mdl-37159678

ABSTRACT

A very common and effective way for investigating future demographics is the study of stage structured models. The focus of this article is to propose a modified model to study the impact of population harvesting on their juvenile and adult stages, and analyze the dynamical properties from both qualitative and numerical perspective. It studies single species stage structured model with linear harvesting on juvenile group and Michaelis-Menten type harvesting on adult group. We exploit general ideas in mathematical modeling process to study the dynamical properties and their biological, ecological, and economic implications. It discusses that bi-stability phenomena may exist, global asymptotic stability at boundary equilibrium points and internal equilibrium points are investigated from construction of suitable Lyapunov and Dulac functions. It has been observed that a suitable linear harvesting on juvenile population can feasibly be carry out along with Michaelis-Menten type harvesting on adult population without endangering extinction of any group of population.

19.
J Genet Eng Biotechnol ; 21(1): 43, 2023 Apr 07.
Article in English | MEDLINE | ID: mdl-37024763

ABSTRACT

BACKGROUND: MicroRNAs (miRNAs) are small endogenous RNAs with an approximate length of 18-22 nucleotides and involved in the regulation of gene expression in transcriptional or post-transcriptional levels. They were found to be associated with leaf morphogenesis, flowering time, vegetative phase change, and response to environmental cues in plants, where they act as a critical regulatory factor. The nature of high conservancy of plant miRNAs within the plant species made it possible to detect the conserved miRNAs by computational approaches. Expressed Sequence Tags (EST) based comparative genomic approaches provide advantages over wet lab approaches as it is convenient, easy to carry out and less time consuming. EST-based in silico approach can unravel new conserved miRNAs in plants, even when the complete genome sequence is not available. RESULTS: To identify the novel miRNAs, a total of 46,865 ESTs from Jatropha curcas were searched for homology to all available 6746 mature miRNAs of plant eudicotyledons. Finally, we ended up with 12 novel miRNAs in Jatropha that range from 18 to 19 nucleotides where their respective precursor miRNAs had 54.11-71.76% (A + U) content. The putative miRNAs belong to 12 individual miRNA family and most of them have higher (A + U) content ranging from 47.36 to 77.77% than their respective miRNA homologs. Many of the target genes by the newly identified miRNAs were associated with plant growth and development, stress response, defense and hormone signaling, and oil synthesis pathways. CONCLUSION: These findings have the potential to speed up miRNA identification and expand our understanding of miRNA functions in J. curcas.

20.
J Phys Chem B ; 127(17): 3861-3869, 2023 May 04.
Article in English | MEDLINE | ID: mdl-37096986

ABSTRACT

Time-resolved fluorescence anisotropy (FA) uses the fluorophore depolarization rate to report on rotational diffusion, conformation changes, and intermolecular interactions in solution. Although FA is a rapid, sensitive, and nondestructive tool for biomolecular interaction studies, the short (∼ns) fluorescence lifetime of typical dyes largely prevents the application of FA on larger macromolecular species and complexes. By using triplet shelving and recovery of optical excitation, we introduce optically activated delayed fluorescence anisotropy (OADFA) measurements using sequential two-photon excitation, effectively stretching fluorescence anisotropy measurement times from the nanosecond scale to hundreds of microseconds. We demonstrate this scheme for measuring slow depolarization processes of large macromolecular complexes, derive a quantitative rate model, and perform Monte Carlo simulations to describe the depolarization process of OADFA at the molecular level. This setup has great potential to enable future biomacromolecular and colloidal studies.

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