Electric field-mediated adhesive dynamics of cells inside bio-functionalised microchannels offers important cues for active control of cell-substrate adhesion.

Adhesive dynamics of cells plays a critical role in determining different biophysical processes orchestrating health and disease in living systems. While the rolling of cells on functionalised substrates having similarity with biophysical pathways appears to be extensively discussed in the literature, the effect of an external stimulus in the form of an electric field on the same remains underemphasized. Here, we bring out the interplay of fluid shear and electric field on the rolling dynamics of adhesive cells in biofunctionalised micro-confinements. Our experimental results portray that an electric field, even restricted to low strengths within the physiologically relevant regimes, can significantly influence the cell adhesion dynamics. We quantify the electric field-mediated adhesive dynamics of the cells in terms of two key parameters, namely, the voltage-altered rolling velocity and the frequency of adhesion. The effect of the directionality of the electric field with respect to the flow direction is also analysed by studying cellular migration with electrical effects acting both along and against the flow. Our experiment, on one hand, demonstrates the importance of collagen functionalisation in the adhesive dynamics of cells through micro channels, while on the other hand, it reveals how the presence of an axial electric field can lead to significant alteration in the kinetic rate of bond breakage, thereby modifying the degree of cell-substrate adhesion and quantifying in terms of the adhesion frequency of the cells. Proceeding further forward, we offer a simple theoretical explanation towards deriving the kinetics of cellular bonding in the presence of an electric field, which corroborates favourably with our experimental outcome. These findings are likely to offer fundamental insights into the possibilities of local control of cellular adhesion via electric field mediated interactions, bearing critical implications in a wide variety of medical conditions ranging from wound healing to cancer metastasis.

Damped Oscillatory Dynamics of a Drop Impacting over Oil-Infused Slippery Interfaces─Does the Oil Viscosity Slow it Down?

A liquid drop impacting on a soft surface is known to exhibit fascinating dynamics that is distinctive from its bounce-back atop a rigid surface. However, while the early spreading of the drop subsequent to its immediate impact with a lubricating liquid layer appears to be reasonably well understood, the later events of retraction and eventual stabilization appear to be poorly addressed. Here, we bring out the nontrivial confluence of the solid substrate wettability and the liquid layer viscosity toward modulating the post-collision dynamics of an impinging liquid drop on a viscous oil-infused surface during its later phase of settlement before arriving at an equilibrium state. Our results reveal that despite an intuitive analogy with the classical phenomenon of damped oscillation, the drop, during its later stages of motion, undergoes dynamical events that may be nontrivially dictated by not only the relative viscosity of the impacting drop and the liquid layer but also the intrinsic wettability of the solid substrate, governing its post-impact settlement via a sequel of spreading-retraction cycles. As a consequence, the viscous liquid layer, instead of providing additional damping, may nonintuitively reduce the effective viscous dissipation so as to hasten the drop's final settlement. These results may turn out to be critical in designing engineered surfaces for tuning the movement of drops in a preferential pathway, bearing decisive implications in the functionalities of liquid lenses, inkjet printing, spray coating and cooling, and several other emerging applications in the realm of lubricated fluidic interfaces.

Acute exposure to a neem based biopesticide and mahua oil cake changes haemocyte parameters in freshwater crab, Varuna litterata (Decapoda, Crustacea).

Present study aims to evaluate the immunotoxic effects of two biopesticides, Nimbecidine Plus (a neem biopesticide) and mahua oil cake (MOC) on the haemocyte populations of a freshwater crab, Varuna litterata after acute exposure. Four-day static renewal bioassay test was performed where sixteen healthy adult male crabs were exposed to 96-h LC50 values of Nimbecidine Plus (0.006284 ppt) and MOC aqueous extract (7.631 ppt) separately in the laboratory condition. Control groups were maintained throughout the experimental period without any biopesticide exposure. Various haemocyte parameters such as total count (THC), differential count (DHC), haemocyte density, cytomorphological anomalies and reactive oxygen species (ROS) were measured in the biopesticides-exposed and control crabs after 24, 48, 72, and 96 h of exposure. After treatment with Nimbecidine Plus and MOC, several cytomorphological deformities (cytoplasmic and nuclear membrane disintegration, chromatin condensation, pyknosis, karyorrhexis, karyolysis, nuclear vacuolation, altered cell shape, cellular coagulation, cytoplasmic discharge, vacuolation) were observed in hyalinocytes, small granule haemocytes and large granule haemocytes with modulation of their relative percentages at different exposure times. THC, DHC, haemocyte density and ROS levels were significantly altered (p < 0.05) in biopesticides-exposed crabs at different exposure periods. The toxicity of both biopesticides did not persist throughout the entire exposure time. Nimbecidine Plus exhibited nonlinear toxic impacts on different haemocyte parameters at initial, mid and higher exposure periods whereas MOC showed linear toxic effects mostly at initial exposure time. In comparison to MOC, Nimbecidine Plus showed higher immunotoxic effects in V. litterata. Outcome of this experiment might provide useful information to understand the immune responses of V. litterata against biopesticide toxicity.

Targeting Magnetic Nanoparticles in Physiologically Mimicking Tissue Microenvironment.

Magnetic nanoparticles as drug carriers, despite showing immense promises in preclinical trials, have remained to be only of limited use in real therapeutic practice primarily due to unresolved anomalies concerning their grossly contrasting controllability and variability in performance in artificial test benches as compared to human tissues. To circumvent the deficits of reported in vitro drug testing platforms that deviate significantly from the physiological features of the living systems and result in this puzzling contrast, here, we fabricate a biomimetic microvasculature in a flexible tissue phantom and demonstrate distinctive mechanisms of magnetic-field-assisted controllable penetration of biocompatible iron oxide nanoparticles across the same, exclusively modulated by tissue deformability, which has by far remained unraveled. Our experiments deciphering the transport of magnetic nanoparticles in a blood analogue medium unveil a decisive interplay of the flexibility of the microvascular pathways, magnetic pull, and viscous friction toward orchestrating the optimal vascular penetration and targeting efficacy of the nanoparticles in colorectal tissue-mimicking bioengineered media. Subsequent studies with biological cells confirm the viability of using localized magnetic forces for aiding nanoparticle penetration within cancerous lesions. We establish nontrivially favorable conditions to induce a threshold force for vascular rupture and eventual target of the nanoparticles toward the desired extracellular site. These findings appear to be critical in converging the success of in vitro trials toward patient-specific targeted therapies depending on personalized vascular properties obtained from medical imaging data.

Effects of salinity stress on antioxidant status and inflammatory responses in females of a "Near Threatened" economically important fish species Notopterus chitala: a mechanistic approach.

In the present study, acute stress responses of adult female Notopterus chitala were scrutinized by antioxidant status and inflammation reaction in the gill and liver at five different salinity exposures (0, 3, 6, 9, 12 ppt). Oxidative defense was assessed by determining superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase, and glutathione reductase activities, while malondialdehyde (MDA), glutathione, and xanthine oxidase levels were determined as indicators of oxidative load. Pro-inflammatory cytokines (IL-1ß, IL-6, IL-10, and TNFα) and caspase 1 levels were also analyzed. Expression levels of transcription factors (NRF2 and NF-κB) and molecular chaperons (HSF, HSP70, and HSP90) were estimated to evaluate their relative contribution to overcome salinity stress. MDA showed a significant (P < 0.05) increase (gill, + 25.35-90.14%; liver, + 23.88-80.59%) with salinity; SOD (+ 13.72-45.09%) and CAT (+ 12.73-33.96%) exhibited a sharp increase until 9 ppt, followed by a decrease at the highest salinity (12 ppt) (gill, - 3.92%; liver, - 2.18%). Levels of cytokines were observed to increase (+ 52.8-127.42%) in a parallel pattern with increased salinity. HSP70 and HSP90 expressions were higher in gill tissues than those in liver tissues. NRF2 played pivotal role in reducing salinity-induced oxidative load in both the liver and gills. Serum cortisol and carbonic anhydrase were measured and noted to be significantly (P < 0.05) upregulated in salinity stressed groups. Gill Na+-K+-ATPase activity decreased significantly (P < 0.05) in fish exposed to 6, 9, and 12 ppt compared to control. Present study suggests that a hyperosmotic environment induces acute oxidative stress and inflammation, which in turn causes cellular death and impairs tissue functions in freshwater fish species such as Notopterus chitala.

Portable, handheld, and affordable blood perfusion imager for screening of subsurface cancer in resource-limited settings.

Precise information on localized variations in blood circulation holds the key for noninvasive diagnostics and therapeutic assessment of various forms of cancer. While thermal imaging by itself may provide significant insights on the combined implications of the relevant physiological parameters, viz. local blood perfusion and metabolic balance due to active tumors as well as the ambient conditions, knowledge of the tissue surface temperature alone may be somewhat inadequate in distinguishing between some ambiguous manifestations of precancer and cancerous lesions, resulting in compromise of the selectivity in detection. This, along with the lack of availability of a user-friendly and inexpensive portable device for thermal-image acquisition, blood perfusion mapping, and data integration acts as a deterrent against the emergence of an inexpensive, contact-free, and accurate in situ screening and diagnostic approach for cancer detection and management. Circumventing these constraints, here we report a portable noninvasive blood perfusion imager augmented with machine learning-based quantitative analytics for screening precancerous and cancerous traits in oral lesions, by probing the localized alterations in microcirculation. With a proven overall sensitivity >96.66% and specificity of 100% as compared to gold-standard biopsy-based tests, the method successfully classified oral cancer and precancer in a resource-limited clinical setting in a double-blinded patient trial and exhibited favorable predictive capabilities considering other complementary modes of medical image analysis as well. The method holds further potential to achieve contrast-free, accurate, and low-cost diagnosis of abnormal microvascular physiology and other clinically vulnerable conditions, when interpreted along with complementary clinically evidenced decision-making perspectives.

Frugal Approach toward Developing a Biomimetic, Microfluidic Network-on-a-Chip for In Vitro Analysis of Microvascular Physiology.

Several disease conditions, such as cancer metastasis and atherosclerosis, are deeply connected with the complex biophysical phenomena taking place in the complicated architecture of the tiny blood vessels in human circulatory systems. Traditionally, these diseases have been probed by devising various animal models, which are otherwise constrained by ethical considerations as well as limited predictive capabilities. Development of an engineered network-on-a-chip, which replicates not only the functional aspects of the blood-carrying microvessels of human bodies, but also its geometrical complexity and hierarchical microstructure, is therefore central to the evaluation of organ-assist devices and disease models for therapeutic assessment. Overcoming the constraints of reported resource-intensive fabrication techniques, here, we report a facile, simple yet niche combination of surface engineering and microfabrication strategy to devise a highly ordered hierarchical microtubular network embedded within a polydimethylsiloxane (PDMS) slab for dynamic cell culture on a chip, with a vision of addressing the exclusive aspects of the vascular transport processes under medically relevant paradigms. The design consists of hierarchical complexity ranging from capillaries (∼80 µm) to large arteries (∼390 µm) and a simultaneous tuning of the interfacial material chemistry. The fluid flow behavior is characterized numerically within the hierarchical network, and a confluent endothelial layer is realized on the inner wall of microfluidic device. We further explore the efficacy of the device as a vascular deposition assay of circulatory tumor cells (MG-63 osteosarcoma cells) present in whole blood. The proposed paradigm of mimicking an in vitro vascular network in a low-cost paradigm holds further potential for probing cellular dynamics as well as offering critical insights into various vascular transport processes.

PSA detection using label free graphene FET with coplanar electrodes based microfluidic point of care diagnostic device.

Affordable point-of-care (PoC) diagnostic devices enable detection of prostate specific antigen (PSA) in resource limited settings. Despite the advancements in PoC systems, most of the reported methods for PSA detection have unsatisfactory detection limits and are based on labelled assays, requiring multiple reagent flow steps which increases both expenses and inconvenience. Circumventing these constraints, we report here the development and validation of a label free, affordable dielectrophoresis (DEP) based graphene field effect transistor (FET) sensor implemented using coplanar electrodes and integrated uniquely with a compact disc based microfluidic platform along with electronics readout for the estimation of PSA at the point of care. Design of coplanar gate electrode which has not been explored earlier is not a straightforward approach. In fact, it has been observed that there is a non-monotonic dependence of the capture of PSA molecules in the channel region of the FET with varying widths and spacings of the gate electrode. The graphene FET based PoC device with optimized coplanar gate electrode is the only label free analytical system for PSA detection requiring simple operation and achieving a detection limit of 1 pg/ml in serum with a wide dynamic range upto 4 ng/ml and appreciable selectivity against potential interferents like bovine serum albumin (BSA) and human immunoglobulin G (IgG). Further, it has been validated satisfactorily with commercially available existing systems using human serum samples. Moreover, the proposed sensing system lowers the detection limit by three orders of magnitude compared to a recent study on label free PoC device on other cancer biomarkers.

Rapid determination of erythrocyte sedimentation rate (ESR) by an electrically driven blood droplet biosensor.

In healthcare practice, the sedimentation rate of red blood cells (erythrocytes) is a widely used clinical parameter for screening of several ailments such as stroke, infectious diseases, and malignancy. In a traditional pathological setting, the total time taken for evaluating this parameter varies typically from 1 to 2 h. Furthermore, the volume of human blood to be drawn for each test, following a gold standard laboratory technique (alternatively known as the Westergren method), varies from 4 to 5 ml. Circumventing the above constraints, here we propose a rapid (∼1 min) and highly energy efficient method for the simultaneous determination of hematocrit and erythrocyte sedimentation rate (ESR) on a microfluidic chip, deploying electrically driven spreading of a tiny drop of blood sample (∼8 µl). Our unique approach estimates these parameters by correlating the same with the time taken by the droplet to spread over a given radius, reproducing the results from more elaborate laboratory settings to a satisfactory extent. Our novel methodology is equally applicable for determining higher ranges of ESR such as high concentration of bilirubin and samples corresponding to patients with anemia and patients with some severe inflammation. Furthermore, the minimal fabrication steps involved in the process, along with the rapidity and inexpensiveness of the test, render the suitability of the strategy in extreme point-of-care settings.

Enzymatic, non enzymatic antioxidants and glucose metabolism enzymes response differently against metal stress in muscles of three fish species depending on different feeding niche.

Current study aims to determine difference in metal accumulation pattern in muscle of Liza parsia (pelagic, omnivore), Amblypharyngodon mola (surface feeder, herbivore) and Mystus gulio (benthic, carnivore) depending on their niche and feeding habit and how it affects the endogenous antioxidants and glucose metabolism in fish muscle. Fishes were collected from Malancha, Diamond Harbour and Chandanpiri, West Bengal, India. Concentrations of lead, zinc, cadmium, chromium were measured in water, sediment and fish muscle. Metal pollution index (MPI) and bioconcentration factor (BCF) was calculated to evaluate the ability of fish to accumulate specific metals in muscle tissue from the aquatic environment. Metal concentrations were found significantly higher (P < 0.05) in water, sediment, fish muscles from Malancha than Chandanpiri and Diamond Harbour. L. parsia (MPI: 0.4-1.6) showed highest metal deposition in their muscle followed by A. mola (MPI: 0.37-1.38) and M. gulio (MPI: 0.2-1.2). Malondealdehyde, superoxide dismutase, catalase, glutathione S transferase, glutathione reductase and cortisol levels increased in case of L. parsia from Malancha and Chandanpiri. Succinate dehydrogenase, lactate dehydrogenase, Ca+2 ATPase and cytochrome C oxidase levels were significantly (P < 0.05) lower at Malancha and Chandanpiri than Diamond Harbour. Heat shock protein (HSP70) expression was significantly (P < 0.05) higher in all fish species at Malancha followed by Chandanpiri and Diamond Harbour. Glucose, glycogen, hexokinase, phosphofructokinase and glycogen phosphorylase levels varied between sites and selected fish species. Serum cortisol level was measured and found to be the highest in L. parsia from Malancha (2.94 ± 0.12 ng/ml) and the lowest in M. gulio from Diamond Harbour (0.7 ± 0.05 ng/ml). The results indicate that metal toxicity alters antioxidant levels, oxidative status and energy production in fish in species specific manner. Our results also indicate that Mystus has the highest degree of adaptability in response to metal toxicity possibly due to its specific food habit and niche position. Therefore, it can be concluded that maintenance of oxidative and metabolic status to combat metal-induced oxidative load will be helpful for the fishes to acquire better resistance under such eco-physiological stress. Alteration of niche and interactive segregation in aquatic organism may be one of the key modulator of resistance against such stress.

Effectiveness of melatonin to restore fish brain activity in face of permethrin induced toxicity.

Present study demonstrates permethrin induced oxidative damage in fish brain and explores effectiveness of melatonin to ameliorate brain function. Adult female Notopterus notopterus were exposed to nominal permethrin concentrations at 1/20th (0.34 µg/l) and 1/10th (0.68 µg/l) of LC50 for 15 days. The measured permethrin concentrations using gas chromatography (GC-ECD) were 0.28 µg/l and 0.57 µg/l, respectively. Some fish were sacrificed to collect brain tissue after 15 days of exposure. Remaining fish from both groups were administered exogenous melatonin (50 µg/kg, 100 µg/kg body weight) for 7 days and brain tissues were collected. Brain enzymes, ntioxidant factors, HSP70, HSP90, nuclear factor-kappa binding (NFkB), melatonin receptor (MT1R) proteins were measured. Permethrin treatment significantly (P < 0.05) decreased the levels of glutathione and brain enzymes. Malondialdehyde (MDA), xanthine oxidase (XO), HSPs increased at each concentration of permethrin. However, superoxide dismutase, glutathione s-transferase levels increased at low permethrin concentration followed by sharp decrease at higher concentration. Expression of NFkB and MT1R increased significantly (P < 0.05). Melatonin administration reinstated activity of brain enzymes, reduced MDA, XO levels and modulated HSPs. Melatonin also increased expression of NFkB and MT1R. Exogenous melatonin improves oxidative status in permethrin stressed fish brain. Melatonin modulates expression of HSPs that enables brain to become stress tolerant and survive by initiating NFkB translocation. Melatonin could act through melatonin receptor protein to induce synthesis of antioxidant proteins. Therefore the study successfully evaluates the potential of melatonin application for better culture and management of fish against pesticide toxicity.

High Temperature Durability of Oleoplaned Slippery Copper Surfaces.

Slippery surfaces, inspired by the functionality of trapping interfaces of specialized leaves of pitcher plants, have been widely used in self-cleaning, anti-icing, antifrost, and self-healing surfaces. They can be fabricated on metallic surfaces as well, presenting a more durable and low-maintenance anticorrosive surface on metals. However, the lack of studies on the durability of these slippery surfaces at high temperature prohibits their practical deployment in real industrial applications where thermal effects are critical and high temperature conditions are inevitable. We present here a unique fabrication technique of a copper-based oleoplaned slippery surface that has been tested for high temperature durability under repeated thermal cycles. Their slipperiness at high temperatures has also been tested in the absence of the Leidenfrost effect. Our findings suggest that these new substrates can be used for fabricating low maintenance surfaces for high temperature applications or even where the surface undergoes repeated thermal cycles like heat exchanger pipes, utensils, engine casings, and outdoor metallic structures.

Mechanical stress-induced autophagic response: A cancer-enabling characteristic?

Metastasis is the leading cause of cancer mortality. Throughout the cascade of metastasis, cancer cells are exposed to both chemical and mechanical cues which influence their migratory behavior and survival. Mechanical forces in the milieu of cancer may arise due to excessive growth of cells in a confinement as in case of solid tumors, interstitial flows within tumors and due to blood flow in the vasculature as in case of circulating tumor cells. The focus of this review is to highlight the mechanical forces prevalent in the cancer microenvironment and discuss the impact of mechanical stresses on cancer progression, with special focus on mechanically induced autophagic response in cancer cells. Autophagy is a cellular homeostatic mechanism that a cell employs not only for recycling of damaged organelles and turnover of proteins involved in cellular migration but also as an adaptive response to survive through unfavourable stresses. Elucidation of the role of mechanically triggered autophagic response may lead to a better understanding of the mechanobiological aspects of metastatic cancer and unravelling the associated signaling mechanochemical pathways may hint at potential therapeutic targets.

Compressive stress-induced autophagy promotes invasion of HeLa cells by facilitating protein turnover in vitro.

Metastasis remains the primary cause of cancer mortality. Throughout the process of metastasis, cancer cells experience mechanical forces, which may turn out to be the key towards their migratory, homeostatic and survival characteristics. However, the influence of compressive stress on the underlying mechanism of cancer cell adaptation during metastasis has remained grossly unexplored. In this study, we have investigated whether compressive force induces autophagy in HeLa cells with potential implications in cellular invasiveness. To this end, we have adopted a simple strategy to create the mechanically-compressed tumor microenvironment, in vitro, by applying appropriate compression to agarose-scaffolded HeLa cell-encapsulated alginate beads. Our findings confirm that compression upregulates autophagy, which promotes paxillin turnover and active MMP-2 secretion, leading to enhanced migration of HeLa cells. We further show that autophagy induction by compression is affected by the phosphorylation of p38 MAPKs, a process that is mediated by intact membrane lipid rafts. Identifying the role of such mechanically triggered cellular responses, guiding crucial processes like cell migration, may lead to better understanding of the mechanobiological aspects of metastatic cancer and unveil potential therapeutic targets.

Evaluation of the acute toxicity of mahua oil cake aqueous extract and its effect on the behavioral responses of the freshwater grapsid crab, Varuna litterata (Fabricius, 1798).

Mortality and behavioral alterations are monitored as the sensitive endpoints in toxicological studies and may be applied as useful biomarkers to assess piscicidal pollution in aquatic environment. Present study assesses acute toxicity of the piscicide, mahua oil cake (MOC), and its effect on the behavioral responses of the freshwater grapsid crab, Varuna litterata, under laboratory conditions. To determine the LC50 values, a 4-day acute static renewal toxicity test was done where 10 adult male crabs (mean length 2.870 ± 0.379 cm; mean weight 9.891 ± 3.951 g) were exposed to different concentrations (1, 5, 10, 15, 20, 25, 30, 35 ppt) of MOC aqueous extract with a control at different exposure periods. The LC50 values are 19.109 ppt for 24 h, 16.052 ppt for 48 h, 11.827 ppt for 72 h, and 7.631 ppt for 96 h. The high LC50 values indicate less sensitivity of this crab to the MOC extract than other aquatic animals. MOC extract has toxic effect on the mouthparts activity, whirling motion of water current producing activity, froth releasing activity, aggregation, balance and coordination actions, medium escaping behavior, locomotor activity, and fecal matter excretion of this crab in different exposure periods. Behavioral responses such as froth releasing activity, aggregation, and medium escaping behavior can be used as biomarkers of MOC pollution in aquatic environment.

Frequency-induced morphology alterations in microconfined biological cells.

Low-intensity therapeutic ultrasound has demonstrated an impetus in bone signaling and tissue healing for decades now. Though this technology is clinically well proven, still there are breaches in studies to understand the fundamental principle of how osteoblast tissue regenerates physiologically at the cellular level with ultrasound interaction as a form of acoustic wave stimuli. Through this article, we illustrate an analysis for cytomechanical changes of cell membrane periphery as a basic first physical principle for facilitating late downstream biochemical pathways. With the help of in situ single-cell direct analysis in a microfluidic confinement, we demonstrate that alteration of low-intensity pulse ultrasound (LIPUS) frequency would physically perturb cell membrane and establish inherent cell oscillation. We experimentally demonstrate here that, at LIPUS resonance near 1.7 MHz (during 1-3 MHz alteration), cell membrane area would expand to 6.85 ± 0.7% during ultrasound exposure while it contracts 44.68 ± 0.8% in post actuation. Conversely, cell cross-sectional area change (%) from its previous morphology during and after switching off LIPUS was reversibly different before and after resonance. For instance, at 1.5 MHz, LIPUS exposure produced 1.44 ± 0.5% expansion while in contrast 2 MHz instigates 1.6 ± 0.3% contraction. We conclude that alteration of LIPUS frequency from 1-3 MHz keeping other ultrasound parameters like exposure time, pulse repetition frequency (PRF), etc., constant, if applied to a microconfined biological single living cell, would perturb physical structure reversibly based on the system resonance during and post exposure ultrasound pulsing. We envision, in the near future, our results would constitute the foundation of mechanistic effects of low-intensity therapeutic ultrasound and its allied potential in medical applications. Graphical Abstract Frequency Dependent Characterization of Area Strain in Cell Membrane by Microfluidic Based Single Cell Analysis.

Electrokinetics with blood.

Microfluidics based lab-on-a-chip technology holds tremendous promises towards point-of-care diagnosis of diseases as well as for developing engineered devices aimed towards replicating the intrinsic functionalities of human bodies as mediated by blood vessel mimicking circulatory networks. While the analysis of transport of blood including its unique cellular constituents has remained to be the focus of many reported studies, a progressive interest on understanding the interplay between electric field and blood flow dynamics has paved a new way towards further developments from scientific engineering as well as clinical viewpoint. Here, we briefly outline the interconnection between electrokinetics and blood flow through micro-capillaries, in an effort to address several challenging propositions in a wide variety of applications encompassing biophysical transport to medical diagnostics. We first present the fundamentals of interaction of electric field with cellular components. In conjunction with the unique rheological features of blood, we show that this interaction may turn out to be compelling for the use of electric fields for transporting blood samples through microfluidic conduits. We discuss the perspectives of both direct current and alternating current electrokinetics in the context of blood flow. In addition, we provide a brief outline of the concerned theoretical developments. We also bring out the relevant biophysical perspectives and focus on applications such as blood plasma separation and separation of circulatory tumor cells. Finally, we attempt to provide a futuristic outlook and envisage the potential of combining electrokinetics with blood microcirculation towards developing futuristic biomimetic microdevices that can replicate a novel control mechanism over micro-circulatory transport in the entire connective network of human bodies. This may effectively pave the way towards the realization of a next-generation medical simulation device, significantly advanced from what is available under the ambit of the state of art technology in the field.

Physiological and behavioural responses to acid and osmotic stress and effects of Mucuna extract in Guppies.

Variation in pH (acidification) and salinity conditions have severe impact at different levels of biological organization in fish. Present study focused to assess the effects of acidification and salinity changes on physiological stress responses at three different levels of function: i) hormonal and oxidative response, ii) osmoregulation and iii) reproduction, in order to identify relevant biomarkers. Second objective of the study was to evaluate the efficacy of plant (Mucuna pruriens) extract for alleviating pH and salinity related stress. Guppies (Poecilia reticulata) were exposed to different pH (6.0, 5.5, 5.0) and salinity (1.5, 3.0, 4.5 ppt) for 7, 14 and 21 days. Following exposure to stress for respective duration, fish were fed diet containing methanol extract of Mucuna seeds (dose 0.80 gm/kg feed) for 7, 14 and 21 days to measure their possible recovery response. Stress hormone (cortisol), hepatic oxidative stress parameters [superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GRd), glutathione peroxidise (GPx), glutathione S-transferase (GST), malondialdehyde (MDA), glutathione (GSH)], gill osmoregulatory response (Na+-K+ATPase activity), sex steroid profiles and mating behaviours (gonopodial thrust and gestation period) were estimated. Cortisol and MDA levels increased with dose and duration of acid and salinity stress, and cortisol levels were higher in males than in females. Effect on Na+-K+ATPase activity was more intense by salinity stress rather than pH induced stress. Both acid and salinity stress reduced sex steroid levels, and mating response was highly affected by both stresses in a dose- and duration-dependent manner. Mucuna treatment reduced stress-induced alteration of cortisol, MDA, Na+-K+ATPase activity and reproductive parameters. Dietary administration of Mucuna seed extract decreased the intensity of environmental stressors at all three functional levels. Mucuna treatment was more effective against salinity stress than acid stress. Thus, cortisol, oxidative stress marker MDA and Na+-K+ATPase could be effective indicators for acid and salinity stress in wild and domestic fish populations. Dietary administration of Mucuna extract may limit the detrimental effects of acidification and salinity variations that are the inevitable outcomes expected under global climate change conditions.

Melatonin ameliorates H2O2-induced oxidative stress through modulation of Erk/Akt/NFkB pathway.

BACKGROUND: Improper control on reactive oxygen species (ROS) elimination process and formation of free radicals causes tissue dysfunction. Pineal hormone melatonin is considered a potent regulator of such oxidative damage in different vertebrates. Aim of the current communication is to evaluate the levels of oxidative stress and ROS induced damage, and amelioration of oxidative status through melatonin induced activation of signaling pathways. Hepatocytes were isolated from adult Labeo rohita and exposed to H2O2 at three different doses (12.5, 25 and 50 µM) to observe peroxide induced damage in fish hepatocytes. Melatonin (25, 50 and 100 µg/ml) was administered against the highest dose of H2O2. Enzymatic and non-enzymatic antioxidants such as malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) was measured spectrophotometrically. Expression level of heat shock proteins (HSP70 and HSP90), HSPs-associated signaling molecules (Akt, ERK, cytosolic and nuclear NFkB), and melatonin receptor was also measured by western blotting analysis. RESULTS: H2O2 induced oxidative stress significantly altered (P < 0.05) MDA and GSH level, SOD and CAT activity, and up regulated HSP70 and HSP90 expression in carp hepatocytes. Signaling proteins exhibited differential modulation as revealed from their expression patterns in H2O2-exposed fish hepatocytes, in comparison with control hepatocytes. Melatonin treatment of H2O2-stressed fish hepatocytes restored basal cellular oxidative status in a dose dependent manner. Melatonin was observed to be inducer of signaling process by modulation of signaling molecules and melatonin receptor. CONCLUSIONS: The results suggest that exogenous melatonin at the concentration of 100 µg/ml is required to improve oxidative status of the H2O2-stressed fish hepatocytes. In H2O2 exposed hepatocytes, melatonin modulates expression of HSP70 and HSP90 that enable the hepatocytes to become stress tolerant and survive by altering the actions of ERK, Akt, cytosolic and nuclear NFkB in the signal transduction pathways. Study also confirms that melatonin could act through melatonin receptor coupled to ERK/Akt signaling pathways. This understanding of the mechanism by which melatonin regulates oxidative status in the stressed hepatocytes may initiate the development of novel strategies for hepatic disease therapy in future.

Hemodynamic shear stress induces protective autophagy in HeLa cells through lipid raft-mediated mechanotransduction.

During metastatic dissemination, cancer cells experience shear stresses in narrow confinements of in vivo vasculature. Such stresses are currently known to influence a gamut of cellular processes. While a host of cells emanating from a primary tumor perish in circulation due to shear, some cells manage to migrate to distant niches and form secondary tumors. Current research focuses on how cancer cells avert such mechanical stresses and adapt themselves in order to survive. This study deals with the autophagic response of cervical cancer cells HeLa and its subline HeLa 229, exposed to physiological shear stresses in vitro, and evaluates its role as a pro-survival mechanism. It also delineates the probable mechanotransduction pathway that is involved in eliciting the stress-adaptive response in cervical cancer cells. Our results show that shear stress of physiological regime elicits protective autophagy in cervical cancer cells as an immediate response and inhibiting the same, leads to early onset of apoptosis. An effort to study the underlying mechanotransduction revealed that autophagy induction by shear stress requires intact lipid rafts which serve as signalling platforms to trigger phosphorylation of p38 mitogen activated protein kinases, leading to autophagy. This study thus gives novel insights into the mechanobiology of cervical cancer and hints at promising therapeutic targets in metastasis, the major cause of cancer mortality.