Semipermeable Barrier Research Articles

Monitoring Blood-Brain Barrier Opening in Rats with a Preclinical Focused Ultrasound System.

The brain has a highly selective semipermeable blood barrier, termed the blood-brain barrier (BBB), which prevents the delivery of therapeutic macromolecular agents to the brain. The integration of MR-guided low-intensity pulsed focused ultrasound (FUS) with microbubble pre-injection is a promising technique for non-invasive and non-toxic BBB modulation. MRI can offer superior soft-tissue contrast and various quantitative assessments, such as vascular permeability, perfusion, and the spatial-temporal distribution of MRI contrast agents. Notably, contrast-enhanced MRI techniques with gadolinium-based MR contrast agents have been shown to be the gold standard for detecting BBB openings. This study outlines a comprehensive methodology involving MRI protocols and animal procedures for monitoring BBB opening in a rat model. The rat model provides the added benefit of jugular vein catheter utilization, which facilitates rapid medication administration. A stereotactic-guided preclinical FUS transducer facilitates the refinement and streamlining of animal procedures and MRI protocols. The resulting methods are characterized by reproducibility and simplicity, eliminating the need for specialized surgical expertise. This research endeavors to contribute to the optimization of preclinical procedures with rat models and encourage further investigation into the modulation of the BBB to enhance therapeutic interventions in neurological disorders.

Disorder-mediated interactions target proteins to specific condensates

Selective compartmentalization of cellular contents is fundamental to the regulation of biochemistry. Although membrane-bound organelles control composition by using a semi-permeable barrier, biomolecular condensates rely on interactions among constituents to determine composition. Condensates are formed by dynamic multivalent interactions, often involving intrinsically disordered regions (IDRs) of proteins, yet whether distinct compositions can arise from these dynamic interactions is not known. Here, by comparative analysis of proteins differentially partitioned by two different condensates, we find that distinct compositions arise through specific IDR-mediated interactions. The IDRs of differentially partitioned proteins are necessary and sufficient for selective partitioning. Distinct sequence features are required for IDRs to partition, and swapping these sequence features changes the specificity of partitioning. Swapping whole IDRs retargets proteins and their biochemical activity to different condensates. Our results demonstrate that IDR-mediated interactions can target proteins to specific condensates, enabling the spatial regulation of biochemistry within the cell.

Quantifying Induced Dipole Effects in Small Molecule Permeation in a Model Phospholipid Bilayer.

The cell membrane functions as a semipermeable barrier that governs the transport of materials into and out of cells. The bilayer features a distinct dielectric gradient due to the amphiphilic nature of its lipid components. This gradient influences various aspects of small molecule permeation and the folding and functioning of membrane proteins. Here, we employ polarizable molecular dynamics simulations to elucidate the impact of the electronic environment on the permeation process. We simulated eight distinct amino-acid side chain analogs within a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine bilayer using the Drude polarizable force field (FF). Our approach includes both unbiased and umbrella sampling simulations. By using a polarizable FF, we sought to investigate explicit dipole responses in relation to local electric fields along the membrane normal. We evaluate molecular dipole moments, which exhibit variation based on their localization within the membrane, and compare the outcomes with analogous simulations using the nonpolarizable CHARMM36 FF. This comparative analysis aims to discern characteristic differences in the free energy surfaces of permeation for the various amino-acid analogs. Our results provide the first systematic quantification of the impact of employing an explicitly polarizable FF in this context compared to the fixed-charge convention inherent to nonpolarizable FFs, which may not fully capture the influence of the membrane dielectric gradient.

Generation and Characterization of hiPSC-Derived Vascularized-, Perfusable Cardiac Microtissues-on-Chip.

In the heart in vivo, vasculature forms a semi-permeable endothelial barrier for selective nutrient and (immune) cell delivery to the myocardium and removal of waste products. Crosstalk between the vasculature and the heart cells regulates homeostasis in health and disease. To model heart development and disease in vitro it is important that essential features of this crosstalk are captured. Cardiac organoid and microtissue models often integrate endothelial cells (ECs) to form microvascular networks inside the 3D structure. However, in static culture without perfusion, these networks may fail to show essential functionality. Here, we describe a protocol to generate an in vitro model of human induced pluripotent stem cell (hiPSC)-derived vascularized cardiac microtissues on a microfluidic organ-on-chip platform (VMToC) in which the blood vessels are perfusable. First, prevascularized cardiac microtissues (MT) are formed by combining hiPSC-derived cardiomyocytes, ECs, and cardiac fibroblasts in a pre-defined ratio. Next, these prevascularized MTs are integrated in the chips in a fibrin hydrogel containing additional vascular cells, which self-organize into tubular structures. The MTs become vascularized through anastomosis between the pre-existing microvasculature in the MT and the external vascular network. The VMToCs are then ready for downstream structural and functional assays and basic characterization. Using this protocol, cardiac MTs can be efficiently and robustly vascularized and perfused within 7 days. In vitro vascularized organoid and MT models have the potential to transition current 3D cardiac models to more physiologically relevant organ models that allow the role of the endothelial barrier in drug and inflammatory response to be investigated. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Generation of VMToC Support Protocol 1: Functional Characterization of VMToC Support Protocol 2: Structural Characterization of VMToC.

Antarctica is less isolated with increasing depth - evidence from pycnogonids

No other group of animals typifies the uniqueness of Antarctic life more than Pycnogonida (sea spiders), with 20% of all known species found in the Southern Ocean, and 64% of these endemic to the Antarctic. Despite nearly 200 years of research into pycnogonids and other benthic phyla in Antarctica, the parameters which drive the distribution and diversity of benthic fauna are still poorly understood. This study aimed to investigate the diversity and connectivity of pycnogonid communities on either side of the Antarctic Polar Front, with an emphasis on the role of water depth, using an occurrence dataset containing 254 pycnogonid species from 2187 sampling locations. At depths shallower than 1000 m, communities to the north and south of the Antarctic Polar Front were distinct, while below this depth this geographic structure disintegrated. The Polar Front, or the expanse of deep ocean it bisects, seemingly acts as a semipermeable barrier to species exchange between well-sampled shallow communities. The less sampled and less understood deep sea appears to be better connected, with high levels of shared species following the northward flow of Antarctic Bottom Water. The exceptionally high diversity and endemism of Antarctic pycnogonids may reflect an apparent competitive advantage in cold waters which leaves them vulnerable to ongoing ocean warming, with increased competition and predation pressures.

Antifungal activity of alginate coatings with essential oil of Mexican oregano incorporated in the stem of tomatoes

SummaryAdding natural antimicrobials such as essential oils (EOs) to alginate coatings on fruits provides an antifungal effect and a semipermeable water vapour barrier. This work aims to evaluate the antifungal effect of Lippia berlandieri S. (Mexican oregano) EO incorporated into alginate to coat tomato stems inoculated with Penicillium commune, Fusarium spp., or Cladosporium herbarum. Alginate films and coatings with EO concentrations (0%, 0.25%, 0.75%, 1%, 2%, or 4% v/v) were prepared to evaluate their properties (physical, mechanical, and water vapour transmission rates) as well as their effect on mould growth on tomato stems for 28 days during refrigerated storage. Mould growth was described with probabilistic and time‐to‐growth predictive models. Two treatment types were assessed (tomato inoculated and coated, TIC, and tomato coated and inoculated, TCI) to provide information about the moment of mould contamination (before or after coating). EO concentration influenced the alginate films' physical, mechanical, and barrier properties. Testing the coatings against moulds revealed varying inhibitory effects. Fusarium spp. exhibited the highest resistance, while C. herbarum was the most susceptible. The predictive models appropriately described the probability of growth or the time‐to‐growth of the studied moulds, identifying critical concentrations of oregano EO needed to inhibit mould growth effectively. The findings suggest that alginate coatings with oregano EO can serve as a natural and effective antifungal treatment for tomatoes, offering insights into optimal concentrations and application methods for extended shelf life.

Evaluation of the Effects of Edible Coatings of Ocimum sanctum and Aloe vera on Jaggery Shelf Life

Background: In western Uttar Pradesh, jaggery is a widespread cottage enterprise based on agriculture, and farmers are forced to sell their product at a lesser price when it is still fresh. Therefore, it was thought to be desirable to create better storage techniques in order to extend its shelf life. Aloe vera, and Ocimum sanctum, often known as "Tulsi," are frequently utilised as antimicrobial food additives because they offer a host of other health advantages in addition to their well-known antibacterial qualities. Because the edible coatings made of these herbs provide a semi-permeable barrier to gases and water vapours, they may prolong the shelf life of jaggery by preventing degradation. Objective: The goal of the current study was to assess the ability of edible coatings of common Indian herbs, such as tulsi and Aloe vera,, to extend the shelf life of jaggery while maintaining attributes that are equal to those of fresh jaggery, in accordance with the guidelines set forth by the Food Safety and Standards Authority of India (FSSAI). Methodology: The physicochemical characteristics, antioxidant activity, total viable count, and antibacterial activity of tulsi-Aloe vera, coated (TAC), Ocimum sanctum, and Aloe vera, (AC) jaggery were assessed and compared with non-coated control. The physicochemical properties were ascertained using standard methodology for measurement of reducing sugars, proteins, phenols, saponins, tannin, alkaloids, and flavonoids. The antimicrobial activity was ascertained by means of the agar double diffusion method. According to established protocol, antioxidant activity was assessed using the DPPH radical scavenging assay and the reducing power assay. Results: According to the research, there is no discernible microbiological deterioration and the edible coatings containing tulsi and Aloe vera, extend the shelf life of jaggery during storage. When compared to uncoated jaggery, coatings were efficient at preventing the growth of both Gramme positive and Gramme negative microorganisms. Over the course of six months, the herb-infused coatings also retained their phenolic, flavonoid, and tannin contents, which improved their anti-oxidant efficacy when compared to the untreated control group.

The Andes as a semi-permeable geographical barrier: Genetic connectivity between structured populations in a widespread spider.

Geographical barriers like mountain ranges impede genetic exchange among populations, promoting diversification. The effectiveness of these barriers in limiting gene flow varies between lineages due to each species' dispersal modes and capacities. Our understanding of how the Andes orogeny contributes to species diversification comes from well-studied vertebrates and a few arthropods and plants, neglecting organisms unable to fly or walk long distances. Some arachnids, such as Gasteracantha cancriformis, have been hypothesized to disperse long distances via ballooning (i.e. using their silk to interact with the wind). Yet, we do not know how the environment and geography shape its genetic diversity. Therefore, we tested whether the Andes contributed to the diversification of G. cancriformis acting as an absolute or semi-permeable barrier to genetic connectivity between populations of this spider at opposite sides of the mountain range. We sampled thousands of loci across the distribution of the species and implemented population genetics, phylogenetic, and landscape genetic analyses. We identified two genetically distinct groups structured by the Central Andes, and a third less structured group in the Northern Andes that shares ancestry with the previous two. This structure is largely explained by the altitude along the Andes, which decreases in some regions, possibly facilitating cross-Andean dispersal and gene flow. Our findings support that altitude in the Andes plays a major role in structuring populations in South America, but the strength of this barrier can be overcome by organisms with long-distance dispersal modes together with altitudinal depressions.

Plant Membrane-On-A-Chip: A Platform for Studying Plant Membrane Proteins and Lipids.

The cell plasma membrane is a two-dimensional, fluid mosaic material composed of lipids and proteins that create a semipermeable barrier defining the cell from its environment. Compared with soluble proteins, the methodologies for the structural and functional characterization of membrane proteins are challenging. An emerging tool for studies of membrane proteins in mammalian systems is a "plasma membrane on a chip," also known as a supported lipid bilayer. Here, we create the "plant-membrane-on-a-chip,″ a supported bilayer made from the plant plasma membranes of Arabidopsis thaliana, Nicotiana benthamiana, or Zea mays. Membrane vesicles from protoplasts containing transgenic membrane proteins and their native lipids were incorporated into supported membranes in a defined orientation. Membrane vesicles fuse and orient systematically, where the cytoplasmic side of the membrane proteins faces the chip surface and constituents maintain mobility within the membrane plane. We use plant-membrane-on-a-chip to perform fluorescent imaging to examine protein-protein interactions and determine the protein subunit stoichiometry of FLOTILLINs. We report here that like the mammalian FLOTILLINs, FLOTILLINs expressed in Arabidopsis form a tetrameric complex in the plasma membrane. This plant-membrane-on-a-chip approach opens avenues to studies of membrane properties of plants, transport phenomena, biophysical processes, and protein-protein and protein-lipid interactions in a convenient, cell-free platform.

ANN multi-layer perceptron for prediction of blood–brain barrier permeable compounds for central nervous system therapeutics

Endothelial cells produce a semipermeable barrier known as the blood–brain barrier (BBB) to keep undesired chemicals out of the central nervous system (CNS). However, this barrier also restricts the exploration of potential new medications due to insufficient exposure. To address this challenge, machine learning (ML) algorithms can be useful to predict the BBB permeability of chemical compounds. Support vector machines, continuous neural networks, and deep learning approaches have been used to identify compounds that can penetrate the BBB. However, predicting BBB permeability based solely on chemical structure can be difficult. In the current research, we developed an ML model using a large dataset to predict BBB permeability, which could be used for early-stage drug screening of potential CNS medications. Our artificial neural network ANN algorithm exhibited an accuracy of 0.94, specificity of 0.83, sensitivity of 0.97, AUC of 0.96, and MCC of 0.83. These metrics suggest that our model has a high accuracy rate in predicting BBB permeability and therefore has the potential to advance drug discovery efforts in the CNS. This study’s outcomes demonstrate the potential for ML models to predict BBB permeability accurately, aiding in the identification of new CNS therapeutic options. Communicated by Ramaswamy H. Sarma

Diffusion across particle-laden interfaces in Pickering droplets.

Emulsions stabilized by nanoparticles, known as Pickering emulsions, exhibit remarkable stability, which enables applications ranging from encapsulation, to advanced materials, to chemical conversion. The layer of nanoparticles at the interface of Pickering droplets is a semi-permeable barrier between the two liquid phases, which can affect the rate of release of encapsulates, and the interfacial transfer of reactants and products in biphasic chemical conversion. A gap in our fundamental understanding of diffusion in multiphase systems with particle-laden interfaces currently limits the optimal development of these applications. To address this gap, we developed an experimental approach for in situ, real-time quantification of concentration fields in Pickering droplets in a Hele-Shaw geometry and investigated the effect of the layer of nanoparticles on diffusion of solute across a liquid-liquid interface. The experiments did not reveal a significant hindrance on the diffusion of solute across an interface densely covered by nanoparticles. We interpret this result using an unsteady diffusion model to predict the spatio-temporal evolution of the concentration of solute with a particle-laden interface. We find that the concentration field is only affected in the immediate vicinity of the layer of particles, where the area available for diffusion is affected by the particles. This defines a characteristic time scale for the problem, which is the time for diffusion across the layer of particles. The far-field concentration profile evolves towards that of a bare interface. This localized effect of the particle hindrance is not measurable in our experiments, which take place over a much longer time scale. Our model also predicts that the hindrance by particles can be more pronounced depending on the particle size and physicochemical properties of the liquids and can ultimately affect performance in applications.

Vascular development, remodeling and maturation.

The development of the vascular system is crucial in supporting the growth and health of all other organs in the body, and vascular system dysfunction is the major cause of human morbidity and mortality. This chapter discusses three successive processes that govern vascular system development, starting with the differentiation of the primitive vascular system in early embryonic development, followed by its remodeling into a functional circulatory system composed of arteries and veins, and its final maturation and acquisition of an organ specific semi-permeable barrier that controls nutrient uptake into tissues and hence controls organ physiology. Along these steps, endothelial cells forming the inner lining of all blood vessels acquire extensive heterogeneity in terms of gene expression patterns and function, that we are only beginning to understand. These advances contribute to overall knowledge of vascular biology and are predicted to unlock the unprecedented therapeutic potential of the endothelium as an avenue for treatment of diseases associated with dysfunctional vasculature.

Development of an automaton recognizer of tissue pathologies caused by Chlamydia infection

Chlamydia belongs to such diseases, in which the blood-brain barrier permeability is disturbed leading to degenerative changes of brain cells and development of neurological symptoms in animals. Chlamydiae disrupt the barrier function of endothelium, which inside blood vessels forms a semipermeable barrier between the contents of vessels and surrounding tissues. As a result of this process, part of the endothelial cytes slough into the lumen of blood vessels and destroying, contributes to the generalization of infection in the body. Chlamydia infection affecting mammals and birds is caused by antigenically related microorganisms from the Chlamydiacea family. Under animal chlamydia a whole range of diseases is considered, which, due to their polymorphic nature, cannot be united by a specific symptom complex, and sometimes affect all systems and organs. Due to the lack of organotropism and host specificity in different representatives of chlamydiae, the clinic of chlamydiae is extremely diverse. As a diagnosis of this disease, as well as the accuracy of the results, the histological method of examination of pathological material is used. In the process of the conducted research qualitative and quantitative characteristics of pathomorphological changes of cellular structures were determined, with the identification of indicators. All pathological processes occurring in the cell of the organism we have divided into three main groups: cell membrane pathology, cell membrane pathology, and Mitochondria pathology. We developed a decision support module for preliminary diagnosis of pathologies. A truth table of the relationship between indicators and pathology groups was developed. Minimization of disjunctive normal form was carried out using finite automata theory.

Lamellipodia dynamics and microrheology in endothelial cell paracellular gap closure

Vascular endothelial cells (ECs) form a semipermeable barrier separating vascular contents from the interstitium, thereby regulating the movement of water and molecular solutes across small intercellular gaps, which are continuously forming and closing. Under inflammatory conditions, however, larger EC gaps form resulting in increased vascular leakiness to circulating fluid, proteins, and cells, which results in organ edema and dysfunction responsible for key pathophysiologic findings in numerous inflammatory disorders. In this study, we extend our earlier work examining the biophysical properties of EC gap formation and now address the role of lamellipodia, thin sheet-like membrane projections from the leading edge, in modulating EC spatial-specific contractile properties and gap closure. Micropillars, fabricated by soft lithography, were utilized to form reproducible paracellular gaps in human lung ECs. Using time-lapse imaging via optical microscopy, rates of EC gap closure and motility were measured with and without EC stimulation with the barrier-enhancing sphingolipid, sphingosine-1-phosphate. Peripheral ruffle formation was ubiquitous during gap closure. Kymographs were generated to quantitatively compare the lamellipodia dynamics of sphingosine-1-phosphate-stimulated and -unstimulated ECs. Utilizing atomic force microscopy, we characterized the viscoelastic behavior of EC lamellipodia. Our results indicate decreased stiffness and increased liquid-like behavior of expanding lamellipodia compared with regions away from the cellular edge (lamella and cell body) during EC gap closure, results in sync with the rapid kinetics of protrusion/retraction motion. We hypothesize this dissipative EC behavior during gap closure is linked to actomyosin cytoskeletal rearrangement and decreased cross-linking during lamellipodia expansion. In summary, these studies of the kinetic and mechanical properties of EC lamellipodia and ruffles at gap boundaries yield insights into the mechanisms of vascular barrier restoration and potentially a model system for examining the druggability of lamellipodial protein targets to enhance vascular barrier integrity.

The Blood-brain Barrier, a Key Bridge to Treat Neurodegenerative Diseases.

As a highly selective and semi-permeable barrier that separates the circulating blood from the brain and central nervous system (CNS), the blood-brain barrier (BBB) plays a critical role in the onset and treatment of neurodegenerative diseases (NDs). To delay or reverse the NDs progression, the dysfunction of BBB should be improved to protect the brain from harmful substances. Simultaneously, a highly efficient drug delivery across the BBB is indispensable. Here, we summarized several methods to improve BBB dysfunction in NDs, including knocking out risk geneAPOE4, regulating circadian rhythms, restoring the gut microenvironment, and activating the Wnt/β-catenin signaling pathway. Then we discussed the advances in BBB penetration techniques, such as transient BBB opening, carrier-mediated drug delivery, and nasal administration, which facilitates drug delivery across the BBB. Furthermore, various in vivo and in vitro BBB models and research methods related to NDs are reviewed. Based on the current research progress, the treatment of NDs in the long term should prioritize the integrity of the BBB. However, a treatment approach that combines precise control of transient BBB permeability and non-invasive targeted BBB drug delivery holds profound significance in improving treatment effectiveness, safety, and clinical feasibility during drug therapy. This review involves the cross application of biology, materials science, imaging, engineering and other disciplines in the field of BBB, aiming to provide multi-dimensional research directions and clinical ideas for the treating NDs.

Hypoxia-Induced miR-101 Impairs Endothelial Barrier Integrity Through Altering VE-Cadherin and Claudin-5.

Stroke is a life-threatening medical condition across the world that adversely affects the integrity of the blood-brain barrier (BBB). The brain microvascular endothelial cells are the important constituent of the BBB. These cells line the blood vessels and form a semipermeable barrier. Disruptions in adherens junction and tight junction proteins of brain microvascular endothelial cells compromise the integrity of BBB. The Vascular Endothelial (VE)-cadherin is an integral adherens junction protein required for the establishment and maintenance of the endothelial barrier integrity. This study aims to investigate the role of miRNA in hypoxia-induced endothelial barrier disruption. In this study, brain endothelial cells were exposed to hypoxic conditions for different time points. Western blotting, overexpression and knockdown of miRNA, real-time PCR, TEER, and sodium fluorescein assay were used to examine the effect of hypoxic conditions on brain endothelial cells. Hypoxic exposure was validated using HIF-1α protein. Exposure to hypoxic conditions resulted to a significant decrease in endothelial barrier resistance and an increase in sodium fluorescein migration across the endothelial barrier. Reduction in endothelial barrier resistance demonstrated compromised barrier integrity, whereas the increase in migration of sodium fluorescein across the barrier indicated the increase in barrier permeability. The present study revealed microRNA-101 decreases the expression of VE-cadherin and claudin-5 in brain endothelial cells exposed to the hypoxic conditions.

Nomadic ungulate movements under threat: Declining mobility of Mongolian gazelles in the Eastern Steppe of Mongolia

Increasing habitat fragmentation and disturbance threaten long-distance movements of ungulates. While the effects of impermeable barriers on ungulate migrations have been well researched, quantitative evidence for gradual, long-term changes of mobility remains rare.We investigated changes in movement behavior of Mongolian gazelle Procapra gutturosa using GPS tracking data collected from 62 gazelle individuals between 2007 and 2021. We quantified 16-day displacement distances as a metric for long-distance movements before using linear mixed models, generalized additive models and quantile regressions to assess how anthropogenic and environmental factors affected gazelle movement behavior.Long-distance 16-day movements decreased by 36 %, from 142 km in 2007 to 92 km in 2021. Changes in mobility were affected by increasing vehicle numbers in Mongolia, but could not be explained by concurrent changes in other environmental factors like temperature, precipitation or vegetation greenness. Gazelle movement decreased close to roads, and gazelles stayed further away from roads during the snow-free season, when traffic likely is most intense.Conserving landscape permeability is essential for maintaining populations of highly mobile species. Our study provides evidence for a gradual decline in gazelle mobility over fifteen years as a response to increasing anthropogenic impact. The transportation infrastructure permeating the Eastern Steppe does not pose physical barriers, yet our findings suggest that increasing traffic volume may create semipermeable barriers to gazelle movement. As human activity is increasing, interactions between ungulates and vehicle traffic need to be closely monitored to identify and mitigate semipermeable barrier effects before landscape permeability is severely altered.

Propofol Suppresses LPS-induced BBB Damage by Regulating miR-130a-5p/ZO-1 Axis

The blood-brain barrier (BBB) is a highly selective semi-permeable barrier that separates circulating blood from the extracellular fluid of the brain and central nervous system, which is crucial for maintaining brain homeostasis. This study aimed to explore the role of propofol in BBB damage and further evaluate the underlying molecular mechanism. Lipopolysaccharide (LPS) was administered to mice to create an in vivo BBB damage mice model. Additionally, hCMEC/D3 cells as brain microvascular endothelial cells (BMECs) were treated with LPS to establish the in vitro BBB damage cell model. Subsequently, propofol was used for the BBB damage model. Evans blue staining and fluorescein sodium were utilized in the in vivo experiments to demonstrate BBB leakage and BBB permeability. Cell counting kit-8 (CCK-8) assay was used to assess cell viability and the trans-endothelial electrical resistance (TEER) value was measured using an epithelial voltmeter. Furthermore, enzyme-linked immunosorbent assay was performed to measure the levels of the inflammatory cytokines such as interleukin-1β (IL-1β) and tumor necrosis factor-alpha (TNF-α). The levels of miR-130a-5p and zonula occludens-1 (ZO-1) in brain tissues and cells were detected using reverse transcription-quantitative polymerase chain reaction, western blot, or immunofluorescence staining. Furthermore, a dual-luciferase reporter assay was used to demonstrate the association between miR-130a-5p and ZO-1. Propofol treatment suppressed BBB leakage, the amount of fluorescein sodium, and the levels of IL-1β and TNF-α in the LPS-induced BBB damage mice model. Meanwhile, propofol treatment increased the TEER value in the LPS-induced hCMEC/D3 cells. Additionally, propofol treatment significantly down-regulated miR-130a-5p and up-regulated ZO-1. More importantly, miR-130a-5p directly targeted ZO-1 and negatively regulated ZO-1 expression in hCMEC/D3 cells. Furthermore, miR-130a-5p mimic partially reversed the effect of propofol on the TEER value and the levels of inflammatory cytokines such as IL-1β and TNF-α in the LPS-induced hCMEC/D3 cells. Propofol suppressed LPS-induced BBB damage by regulating miR-130a-5p/ZO-1 axis. These findings suggested a potentially effective treatment approach for BBB damage.

Optimization of extraction of starch from sweet potato and its application in making edible film

Present study is based on extraction of starch from sweet potato and then using the extracted starch to manufacture edible films. Gelatine and sweet potato starch based various films (1:1, 2:3, 1:3 and 1:5) have been developed and the optimized concentration of the films has been determined on the basis of functional properties. The characterization techniques, XRD, FTIR and SEM were used to identify the structure, bonds present, granular size and shape. The increase in health awareness among consumers has increased the demand in minimally processed and chemical free fruits and vegetables. Fresh, minimally processed produce can be enrobed in edible coverings made from the edible source to increase the nutritional value. Edible coatings films made from starch offers a possible method of increasing post harvest life providing a semi-permeable barrier for gaseous exchange. Such coatings can reduce respiration, water loss and acts as carrier of preservatives and antioxidants.

Phylogeography of the Asian green mussel, Perna viridis: pronounced regional genetic differentiation is explained by major freshwater input into the Bay of Bengal

The Asian green mussel (AGM), Perna viridis, is a mussel of ecological and economic importance throughout much of the tropical and subtropical regions of Asia, from China and Korea in the east to Oman on the Arabian Peninsula in the west. We collated published DNA sequence data and then analysed 467 bp of cytochrome c oxidase I gene (COI) sequence variation from 649 mussels (576 from SE Asia and 73 from India) to test for regional differentiation. Analysis of molecular variation, a haplotype network and a neighbour-joining tree all revealed significant differentiation between mussels from India and those from SE Asia. We observed a greater number of segregating sites (haplotypes) and private haplotypes than expected for Indian mussels compared to SE Asian mussels, based on the respective sample sizes. Tests of neutrality revealed population expansion or a recent selective sweep for only the mussels from India. We suggest that the differentiation of the Indian from the SE Asian mussels is explained by the pronounced seasonal input of freshwater into the northern Bay of Bengal area, which acts as a (semi-permeable) barrier to gene flow between the two regions (i.e., east versus west of the Bay). This suggestion is based on the oceanography of the region and is consistent with the biogeography and reports of genetic breaks in a range of taxa in this region. Further targetted sampling of AGMs from the east coast of India, Bangladesh and Myanmar is required to test this hypothesis, and additional sampling of AGMs from west of India (e.g., Pakistan, Iran, the Arabian Peninsula) will also be informative. Finally, this study is based on the analysis of a single marker (COI) because this is all that is presently available from published sources but we note that the application of new molecular markers such as single nucleotide polymorphisms to newly collected mussel samples will greatly advance our understanding of the AGM genetic discontinuity and its age, and help test its geographical location and its nature (e.g., a steep/shallow cline or a stepped cline).