Electrical signature of heterogeneous human mesenchymal stem cells.

Human mesenchymal stem cells (hMSCs) have gained traction in transplantation therapy due to their immunomodulatory, paracrine, immune-evasive, and multipotent differentiation potential. The inherent heterogeneity of hMSCs poses a challenge for therapeutic treatments and necessitates the identification of robust biomarkers to ensure reproducibility in both in vivo and in vitro experiments. In this study, we utilized dielectrophoresis (DEP), a label-free electrokinetic phenomenon, to investigate the heterogeneity of hMSCs derived from bone marrow (BM) and adipose tissue (AD). The electrical properties of BM-hMSCs were compared to homogeneous mouse fibroblasts (NIH-3T3), human fibroblasts (WS1), and human embryonic kidney cells (HEK-293). The DEP profile of BM-hMSCs differed most from HEK-293 cells. We compared the DEP profiles of BM-hMSCs and AD-hMSCs and found that they have similar membrane capacitances, differing cytoplasm conductivity, and transient slopes. Inducing both populations to differentiate into adipocyte and osteoblast cells revealed that they behave differently in response to differentiation-inducing cytokines. Histology and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analyses of the differentiation-related genes revealed differences in heterogeneity between BM-hMSCs and AD-hMSCs. The differentiation profiles correlate well with the DEP profiles developed and indicate differences in the heterogeneity of BM-hMSCs and AD-hMSCs. Our results demonstrate that using DEP, membrane capacitance, cytoplasm conductivity, and transient slope can uniquely characterize the inherent heterogeneity of hMSCs to guide robust and reproducible stem cell transplantation therapies.

Assessment of ecosystem health of a micro-level Ramsar coastal zone in the Vembanad Lake, Kerala, India.

Health of an ecosystem is very much important as we depend on its goods and services for our existence. Because of this, we need to continuously monitor its health for human benefit and for identifying areas for improvement of our natural systems. The present study tries to assess the condition of a coastal ecosystem within the Vembanad Lake, Kerala, India, using key water quality parameters at micro-level. Principal component analysis identified the minimum required water quality dataset for further analysis and was scored using linear scoring functions. The weighted additive method was used to integrate the individual scores to arrive at a final score representing the ecosystem health. Spline interpolation was applied to develop the ecosystem health map of the study area. Using this method, 35.8% area of the aquatic ecosystem studied was characterized as good, 32.2% as moderate, 26.2% as fair and 5.8% as poor. The assessment results can help the policymakers/managers to make appropriate decisions for the better management of the coastal ecosystems studied. Moreover, this methodology can be replicated for the assessment of coastal regions with similar ecosystem characteristics.

Epizootics of epizootic ulcerative syndrome among estuarine fishes of Kerala, India, under post-flood conditions.

Epizootic ulcerative syndrome (EUS), primarily caused by the water mold Aphanomyces invadans, is an OIE-notifiable disease, having potential impacts on fisheries. We report EUS epizootics among estuarine fishes of Kerala, India, during 2018, under post-flood conditions 3 decades after its primary outbreak. Six fish species (Mugil cephalus, Platycephalus sp., Scatophagus argus, Arius sp., Planiliza macrolepis and Epinephelus malabaricus) were infected, including the first confirmed natural case in E. malabaricus and P. macrolepis. Salinity, surface temperature, dissolved oxygen and pH of resident water during the epizootic were <2 ppt, 25°C, 4.1 ppm and 7.0. The presence of zoonotic bacterial pathogens (Aeromonas veronii, Shewanella putrefaciens, Vibrio vulnificus and V. parahaemolyticus) in tissues of affected fish indicates that EUS-infected fish may pose a public health hazard if not handled properly. Lack of clinical evidence in the region during the last 3 decades, a high number of affected fishes, including 2 new fish species, the severity of skin lesions and very low water salinity (<2 ppt) during the outbreak in contrast to historical water salinity records suggest relatively recent invasion by A. invadans. Phylogenetic analysis based on the internal transcribed spacer region of the rRNA gene showed that the same clone of pathogen has spread across different continents regardless of fish species and ecotypes (fresh/estuarine environments). Altogether, the present study provides baseline data which can be applied in EUS management strategies within brackish-water ecosystems. We recommend strict surveillance and development of sound biosecurity measures against the disease.

Separation of neural stem cells by whole cell membrane capacitance using dielectrophoresis.

Whole cell membrane capacitance is an electrophysiological property of the plasma membrane that serves as a biomarker for stem cell fate potential. Neural stem and progenitor cells (NSPCs) that differ in ability to form neurons or astrocytes are distinguished by membrane capacitance measured by dielectrophoresis (DEP). Differences in membrane capacitance are sufficient to enable the enrichment of neuron- or astrocyte-forming cells by DEP, showing the separation of stem cells on the basis of fate potential by membrane capacitance. NSPCs sorted by DEP need not be labeled and do not experience toxic effects from the sorting procedure. Other stem cell populations also display shifts in membrane capacitance as cells differentiate to a particular fate, clarifying the value of sorting a variety of stem cell types by capacitance. Here, we describe methods developed by our lab for separating NSPCs on the basis of capacitance using several types of DEP microfluidic devices, providing basic information on the sorting procedure as well as specific advantages and disadvantages of each device.

Fabrication of poly (lactic-co-glycolic acid)/gelatin electro spun nanofiber patch containing CaCO3/SiO2 nanocomposite and quercetin for accelerated diabetic wound healing.

An open wound or sore on the bottom of the foot caused by diabetes is known as a diabetic foot ulcer. Preventive measures are essential, including consistent foot care and glycemic management. The dangers associated with diabetic foot ulcers can be reduced via early identification and timely treatment. The risk of foot ulcers and limb amputation increases with age and duration of diabetes. Quercetin contains anti-inflammatory and antioxidant properties. Furthermore, the calcium carbonate/silica (CaCO3/SiO2) nanocomposite has a good anti-inflammatory property due to the presence of calcium, which will aid in wound healing. As a result, combining quercetin (plant based anti-inflammatory drug) and CaCO3/SiO2 nanocomposite will boost the wound healing rate. We have synthesized CaCO3/SiO2 nanocomposite in sol-gel method and characterized using XRD, FTIR and TEM. Cell line tests and the MTT assay revealed that the PLGA/gelatin/CaCO3/SiO2/quercetin patch enhanced the proliferation of cells. Its anti-bacterial efficacy against four major bacterial strains often found in wound locations, as well as its water retention, make it an ideal material for diabetic wound healing. In-vivo trials confirms the enhanced diabetic wound healing potential of the patch.

Synthesis and Characterization of Naringin Functionalized Nano-Hydroxyapatite for Bone Tissue Engineering.

Bone is a unique nanocomposite tissue composed of organic and inorganic materials. Bone grafting is a common surgical method used to improve bone regeneration in dentistry and orthopedic surgery. Because standard therapies have substantial drawbacks, nanomaterials provide alternative options for bone repair. Owing to its high bioactivity, osteoconductivity, biocompatibility, and topography that matches the architecture of real bone, hydroxyapatite nanoparticles (n-HA) are commonly used in bone treatment. We report here the synthesis and characterization of Naringin (NA) functionalized n-HA using HRTEM, FTIR, XRD, and UV-visible spectroscopy. The obtained results indicated that the n-HA can be functionalized with Naringin and they might be used as a bone regenerative material in medical and dental fields.

Water and sediment quality parameters of the restored mangrove ecosystem of Gurupura River and natural mangrove ecosystem of Shambhavi River in Dakshina Kannada, India.

Restoration of Rhizophora mucronata stand in the Gurupura Estuary resulted in improved water and sediment quality parameters. Monthly monitoring from 2011 to 2016 indicates that the restored mangroves grew to a height of 61.49 ± 5.76 cm. Principal component analysis (PCA) showed that increased duration of salinity in the estuary aided the growth of barnacles in planted mangroves which reduced survival by 10%. The United States Environmental Protection Agency ratings revealed that natural mangrove site in Shambavi River exhibit the maximum good water quality rating though dissolved inorganic phosphorous was rated highest due to non-point pollution sources. The pH, salinity, dissolved oxygen, silicate, phosphate, ammonia, and rainfall demonstrated significant seasonal differences (P < 0.001). Mangrove roots and biomes aided in accumulation of clay and significant difference (P < 0.001) was observed yearly. Land use management, efficient waste disposal system along with restoration of diverse mangroves can improve the water quality of estuarine ecosystem.

Photo induced mechanistic activity of GO/Zn(Cu)O nanocomposite against infectious pathogens: Potential application in wound healing.

Treating infection causing microorganisms is one of the major challenges in wound healing. These may gain resistance due to the overuse of conventional antibiotics. A promising technique is antimicrobial photodynamic therapy (aPDT) used to selectively cause damage to infectious pathogenic cells via generation of reactive oxygen species (ROS). We report on biocompatable nanomaterials that can serve as potential photosensitizers for aPDT. GO/Zn(Cu)O nanocomposite was synthesized by co-precipitation method. Graphene Oxide (GO) is known for its high surface to volume ratio, excellent surface functionality and enhanced antimicrobial property. ZnO nanoparticle induces the generation of reactive oxygen species (ROS) under light irradiation and it leads to recombination of electron-hole pair. Nanocomposites of GO and Cu doped ZnO increases visible light absorption and enhances the photocatalytic property. It generates more ROS and increases the bacterial inhibition. GO/Zn(Cu)O nanocomposite was tested against Staphylococcus aureus (S. aureus), Enterococcus faecium (E. faecium), Escherichia coli (E. coli), Salmonella typhi (S. typhi), Shigella flexneri (S. flexneri) and Pseudomonas aeruginosa (P. aeruginosa) by well diffusion method, growth curve, colony count, biofilm formation under both dark and visible light condition. Reactive Oxygen Species assay (ROS), Lactate dehydrogenase leakage (LDH) assay, Protein estimation assay and membrane integrity study proves the mechanism of inhibition of bacteria. Inhibition kinetics shows the sensitivity between bacteria and GO/Zn(Cu)O nanocomposite.

Nanocomposite chitosan film containing graphene oxide/hydroxyapatite/gold for bone tissue engineering.

Recently, polymer based biomaterials are utilized in medical fields including surgical sutures, drug delivery devices, tissue supports and implants for interior bone fixation. However, polymer based implants leads to the formation of bio-films that are highly susceptible to microbial adhesion. In this study, we have fabricated Chitosan/Polyvinyl alcohol/Graphene oxide/Hydroxyapatite/gold films for potential orthopedic application. Graphene oxide/Hydroxyapatite/gold nanocomposite (GO/HAP/Au) was synthesized by simple hydrothermal method and GO/HAP/Au nanocomposite incorporated polymeric film was fabricated using gel casting method. The morphology, phase composition, crystalline structure and chemical state of the nanocomposite were characterized using as XRD, HR-TEM, FE-SEM and FT-IR. The bio-films were found to be biocompatible with mouse mesenchymal cells and it enhanced osteoblast differentiation as evidenced by more alkaline phosphatase activity at the cellular level. Hence, these results suggested that the developed nanocomposites films are osteogenic potential for treating bone and bone-related diseases. Antibacterial analysis of the films shows high inhibition zones against Gram positive and Gram Negative bacteria (Escherichia coli, streptococcus mutans, Staphylococcus aureus and Pseudomonas aeruginosa). Thus, the obtained nanocomposites bio-films are highly biocompatible and it can be used for bone regeneration application.

Studies pertaining to the monitoring of volatile halogenated anaesthetics in breath by proton transfer reaction mass spectrometry.

Post-operative isoflurane has been observed to be present in the end-tidal breath of patients who have undergone major surgery, for several weeks after the surgical procedures. A major new non-controlled, non-randomized, and open-label approved study will recruit patients undergoing various surgeries under different inhalation anaesthetics, with two key objectives, namely (1) to record the washout characteristics following surgery, and (2) to investigate the influence of a patient's health and the duration and type of surgery on elimination. In preparation for this breath study using proton transfer reaction time-of-flight mass spectrometry (PTR-TOF-MS), it is important to identify first the analytical product ions that need to be monitored and under what operating conditions. In this first paper of this new research programme, we present extensive PTR-TOF-MS studies of three major anaesthetics used worldwide, desflurane (CF3CHFOCHF2), sevoflurane ((CF3)2CHOCH2F), and isoflurane (CF3CHClOCHF2) and a fourth one, which is used less extensively, enflurane (CHF2OCF2CHFCl), but is of interest because it is an isomer of isoflurane. Product ions are identified as a function of reduced electric field (E/N) over the range of approximately 80 Td to 210 Td, and the effects of operating the drift tube under 'normal' or 'humid' conditions on the intensities of the product ions are presented. To aid in the analyses, density functional theory (DFT) calculations of the proton affinities and the gas-phase basicities of the anaesthetics have been determined. Calculated energies for the ion-molecule reaction pathways leading to key product ions, identified as ideal for monitoring the inhalation anaesthetics in breath with a high sensitivity and selectivity, are also presented.

PVA/SA/TiO2-CUR patch for enhanced wound healing application: In vitro and in vivo analysis.

Wound healing is a complex multistep process. Wound healing materials should have good antibacterial activity against wound infection causing microbes. Curcumin has effective antimicrobial activity, anti-inflammatory and antioxidant property. Titanium dioxide (TiO2) is a biocompatible, nontoxic material used for many biomedical applications. The Usage of curcumin tagged TiO2 nanoparticles for wound healing activity is promising due to the properties of both curcumin and TiO2. We have synthesized curcumin tagged TiO2 nanoparticles. The synthesized materials are characterized with XRD, FTIR and TEM. TiO2-Cur nanocomposite was incorporated into poly vinyl alcohol (PVA) and sodium alginate (SA) patch. The PVA/SA/TiO2-Cur patch was prepared by gel casting method. Antibacterial efficiency of PVA/SA/TiO2-Cur patch was analyzed. Further, in vivo studies conducted on Wister rats confirmed the enhanced wound healing property of the PVA/SA/TiO2-Cur patch. Our results suggest that this could be an ideal biomaterial for wound dressing applications.

Proton transfer reaction time-of-flight mass spectrometric measurements of volatile compounds contained in peppermint oil capsules of relevance to real-time pharmacokinetic breath studies.

With the growing interest in the use of breath volatiles in the health sciences, the lack of standardization for the sampling and analysis of exhaled breath is becoming a major issue leading to an absence of conformity, reproducibility and reliability in spectrometric measurements. Through the creation of a worldwide 'peppermint consortium', the International Association of Breath Research has set up a task force to deal with this problem. Pharmacokinetic studies are proposed, and a real-time analytical technique that is being used is proton transfer reaction-time-of-flight-mass spectrometry (PTR-ToF-MS). This paper presents details on how the volatile compounds contained in a peppermint oil capsule, and hence on breath, appear in a PTR-ToF-MS. To aid that study, the key volatiles in the headspace of peppermint oil were first identified using gas chromatography-mass spectrometry, notably: menthol, menthone, 1,8-cineole, menthofuran, limonene, α-pinene and ß-pinene. A PTR-ToF-MS analysis of these compounds has been undertaken, divorced from the complexity of the peppermint oil matrix using 'normal' and 'saturated' humidity drift-tube conditions, with the latter used to mimic breath samples, and over a range of reduced electric fields. There are no characteristic product ions that can distinguish monoterpenes and 1,8-cineole, and hence, without pre-separation, a combined washout for these volatiles can only be provided. By operating the drift tube above about 130 Td, there are characteristic product ions for menthone, menthofuran and menthol, namely m/z 155.14 (protonated menthone), m/z 151.11 (protonated menthofuran), m/z 139.15 (loss of H2O from protonated menthol) and m/z 83.09 (a fragment ion, C6H11 +, from menthol). These have been used to monitor, with a high specificity, the temporal profile of these three compounds in breath following the ingestion of a peppermint oil capsule. To aid in the analyses, the proton affinities and gas-phase basicities for the key volatiles investigated have been determined using density functional theory.

Conjoined twins - role of imaging and recent advances.

INTRODUCTION: Conjoined twins are identical twins with fused bodies, joined in utero. They are rare complications of monochorionic twinning. The purpose of this study is to describe the various types of conjoined twins, the role of imaging and recent advances aiding in their management. MATERIAL AND METHODS: This was a twin institutional study involving 3 cases of conjoined twins diagnosed over a period of 6 years from 2010 to 2015. All the 3 cases were identified antenatally by ultrasound. Only one case was further evaluated by MRI. RESULTS: Three cases of conjoined twins (cephalopagus, thoracopagus and omphalopagus) were accurately diagnosed on antenatal ultrasound. After detailed counseling of the parents and obtaining written consent, all the three cases of pregnancy were terminated. Delivery of the viable conjoined twins was achieved without any complications to the mothers, and all the three conjoined twins died after a few minutes. CONCLUSION: Ultrasound enables an early and accurate diagnosis of conjoined twins, which is vital for obstetric management. MRI is reserved for better tissue characterization. Termination of pregnancy when opted, should be done at an early stage as later stages are fraught with problems. Recent advances, such as 3D printing, may aid in surgical pre-planning, thereby enabling successful surgical separation of conjoined twins.

Safety, bioavailability and mechanism of action of nitric oxide to control Bovine Respiratory Disease Complex in calves entering a feedlot.

Bovine Respiratory Disease Complex (BRDc), a multi-factorial disease, negatively impacts the cattle industry. Nitric oxide (NO), a naturally occurring molecule, may have utility controlling incidence of BRDc. Safety, bioavailability, toxicology and tolerance/stress of administering NO to cattle is evaluated herein. Thirteen, crossbred, multiple-sourced, commingled commercial weaned beef calves were treated multiple times intranasally over a 4 week period with either a nitric oxide releasing solution (treatment) or saline (control). Exhaled NO, methemoglobin percent (MetHg) and serum nitrites demonstrated biological availability as a result of treatment. Cortisol levels, tissue nitrites, behavior and gross and macroscopic pathology of organs were all normal. Moreover, preliminary in vitro studies using Mannheimia haemolytica, Infectious Bovine Rhinotracheitis, Bovine Parainfluenza-3 and Bovine Respiratory Syncytial Virus, suggest a potential explanation for the previously demonstrated efficacy for BRDc. These data confirm the bioavailability, safety and lack of residual of NO treatment to cattle, along with the bactericidal and virucidal effects.