Engineered Microbes for Pigment Production Using Waste Biomass

Received: February 04, 2020; Revised: March 08, 2020; Accepted: March 16, 2020.Agri-food waste biomass is the most abundant organic waste and has high valorisation potential for sustainable bioproducts development. These wastes are not only recyclable in nature but are also rich sources of bioactive carbohydrates, peptides, pigments, polyphenols, vitamins, natural antioxidants, etc. Bioconversion of agri-food waste to value-added products is very important towards zero waste and circular economy concepts. To reduce the environmental burden, food researchers are seeking strategies to utilize this waste for microbial pigments production and further biotechnological exploitation in functional foods or value-added products. Microbes are valuable sources for a range of bioactive molecules, including microbial pigments production through fermentation and/or utilisation of waste. Here, we have reviewed some of the recent advancements made in important bioengineering technologies to develop engineered microbial systems for enhanced pigments production using agrifood wastes biomass/by-products as substrates in a sustainable way.MS, VKG and RB acknowledge ERA Chair for Food (By-) Products Valorization Technologies of the Estonian University of Life Sciences (VALORTECH) which has received funding from the European Union’s Horizon 2020 research and innovation program (under grant agreement No. 810630)

Gelatin-based 3D conduits for transdifferentiation of mesenchymal stem cells into Schwann cell-like phenotypes

In this study, gelatin-based 3D conduits with three different microstructures (nanofibrous, macroporous and ladder-like) were fabricated for the first time via combined molding and thermally induced phase separation (TIPS) technique for peripheral nerve regeneration. The effects of conduit microstructure and mechanical properties on the transdifferentiation of bone marrow-derived mesenchymal stem cells (MSCs) into Schwann cell (SC) like phenotypes were examined to help facilitate neuroregeneration and understand material-cell interfaces. Results indicated that 3D macroporous and ladder-like structures enhanced MSC attachment, proliferation and spreading, creating interconnected cellular networks with large numbers of viable cells compared to nanofibrous and 2D-tissue culture plate counterparts. 3D-ladder-like conduit structure with complex modulus of ∼0.4 × 106 Pa and pore size of ∼150 μm provided the most favorable microenvironment for MSC transdifferentiation leading to ∼85% immunolabeling of all SC markers. On the other hand, the macroporous conduits with complex modulus of ∼4 × 106 Pa and pore size of ∼100 μm showed slightly lower (∼65% for p75, ∼75% for S100 and ∼85% for S100β markers) immunolabeling. Transdifferentiated MSCs within 3D-ladder-like conduits secreted significant amounts (∼2.5 pg/mL NGF and ∼0.7 pg/mL GDNF per cell) of neurotrophic factors, while MSCs in macroporous conduits released slightly lower (∼1.5 pg/mL NGF and 0.7 pg/mL GDNF per cell) levels. PC12 cells displayed enhanced neurite outgrowth in media conditioned by conduits with transdifferentiated MSCs. Overall, conduits with macroporous and ladder-like 3D structures are promising platforms in transdifferentiation of MSCs for neuroregeneration and should be further tested in vivo. Statement of Significance This manuscript focuses on the effect of microstructure and mechanical properties of gelatin-based 3D conduits on the transdifferentiation of mesenchymal stem cells to Schwann cell-like phenotypes. This work builds on our recently accepted manuscript in Acta Biomaterialia focused on multifunctional 2D films, and focuses on 3D microstructured conduits designed to overcome limitations of current strategies to facilitate peripheral nerve regeneration. The comparison between conduits fabricated with nanofibrous, macroporous and ladder-like microstructures showed that the ladder-like conduits showed the most favorable environment for MSC transdifferentiation to Schwann-cell like phenotypes, as seen by both immunolabeling as well as secretion of neurotrophic factors. This work demonstrates the importance of controlling the 3D microstructure to facilitate tissue engineering strategies involving stem cells that can serve as promising approaches for peripheral nerve regeneration.US Army Medical Research and Materiel Command (W81XWH-11-1-0700); Stem Cell Biology Fund; Stanley Endowed Chai

Polymorphism and epitope sharing between the alleles of merozoite surface protein-1 of Plasmodium falciparum among Indian isolates

<p>Abstract</p> <p>Background</p> <p>The C-terminal region of merozoite surface protein-1 (MSP-1) is one of the leading candidates for vaccination against the erythrocytic stages of malaria. However, a major concern in the development of MSP-1 based malaria vaccine is the polymorphism observed in different geographical <it>Plasmodium falciparum </it>isolates. To explore whether the sequence heterogeneity of PfMSP-1 leads to variation in naturally acquired anti-MSP-1₁₉antibodies, the present study was undertaken to study PfMSP-1₁₉sequence polymorphism in malaria-endemic villages in eastern India and also carried out a competition enzyme-linked immunosorbent assay using three PfMSP-1₁₉variant forms.</p> <p>Methods</p> <p>The sequence variations in the C-terminal region of PfMSP-1₁₉were determined in a malaria endemic region. Three PfMSP-1₁₉variants were produced in <it>Escherichia coli </it>(PfMSP1₁₉QKNG-L, PfMSP1₁₉EKNG-L and PfMSP1₁₉ETSR-F) and an immunodepletion assay was carried out using the corresponding patients' sera.</p> <p>Results</p> <p>Results revealed predominance of PfMAD20 allele among Indian field isolates. Seven PfMSP-1₁₉variant forms were isolated in a singe geographical location. Three of PfMSP-1₁₉variant forms when expressed in <it>E. coli </it>showed presence of cross-reaction as well as variant specific antibodies in malaria infected patient sera.</p> <p>Conclusion</p> <p>The present study demonstrates the existence of allele specific antibodies in <it>P. falciparum</it>-infected patient sera, however their role in protection requires further investigation. These results thereby, suggest the importance of a multi-allelic PfMSP-1₁₉based vaccine for an effective malaria control.</p

High Throughput Characterization of Adult Stem Cells Engineered for Delivery of Therapeutic Factors for Neuroprotective Strategies

Mesenchymal stem cells (MSCs) derived from bone marrow are a powerful cellular resource and have been used in numerous studies as potential candidates to develop strategies for treating a variety of diseases. The purpose of this study was to develop and characterize MSCs as cellular vehicles engineered for delivery of therapeutic factors as part of a neuroprotective strategy for rescuing the damaged or diseased nervous system. In this study we used mouse MSCs that were genetically modified using lentiviral vectors, which encoded brain-derived neurotrophic factor (BDNF) or glial cell-derived neurotrophic factor (GDNF), together with green fluorescent protein (GFP). Before proceeding with in vivo transplant studies it was important to characterize the engineered cells to determine whether or not the genetic modification altered aspects of normal cell behavior. Different culture substrates were examined for their ability to support cell adhesion, proliferation, survival, and cell migration of the four subpopulations of engineered MSCs. High content screening (HCS) was conducted and image analysis performed. Substrates examined included: poly-L-lysine, fibronectin, collagen type I, laminin, entactin-collagen IV-laminin (ECL). Ki67 immunolabeling was used to investigate cell proliferation and Propidium Iodide staining was used to investigate cell viability. Time-lapse imaging was conducted using a transmitted light/environmental chamber system on the high content screening system. Our results demonstrated that the different subpopulations of the genetically modified MSCs displayed similar behaviors that were in general comparable to that of the original, non-modified MSCs. The influence of different culture substrates on cell growth and cell migration was not dramatically different between groups comparing the different MSC subtypes, as well as culture substrates. This study provides an experimental strategy to rapidly characterize engineered stem cells and their behaviors before their application in longterm in vivo transplant studies for nervous system rescue and repair

Comparison of two ultra-widefield cameras with high image resolution and wider view for identifying diabetic retinopathy lesions

Purpose: To compare the effectiveness of the Optos P200dTx and Zeiss Clarus 50fundus cameras in detecting diabetic retinopathy (DR) lesions. Methods: A cross-sectional study was conducted among 243 patients with clinically diagnoseddiabetesmellituswhowerereferredforaneyeexaminationfromtwotertiary eye care centers in Chennai, India. Patients underwent DR screening based on mydriatic fundal images acquired by both fundal cameras. Fundal images from the two separate devices for each eye were compared based on accurately identified pathological retinal lesions with respect to type and location. Results: When studying lesions of the central retina, they were better identified by the Zeiss Clarus compared with the Optos P200dTx, with six out of eight being statistically significant (P < 0.05). However, lesions of the mid-peripheral retina and peripheral retina were better identified by the Optos P200dTx than the Zeiss Clarus, with three out of eight lesions and five out of eight lesions being statistically significant (P < 0.05), respectively. Based on the color and size of lesions, the Optos P200dTx had a higher chance (59.6%) of missing white lesions than did the Zeiss Clarus (17%) (P < 0.0001). Consequently, small-and medium-sized lesions were missed more by the Optos P200dTx (30.72% and 32.63%, respectively) than the Zeiss Clarus (22.3% and 19.30%, respectively). Conclusions: The capability of detecting or missing a particular DR lesion among diabetics differed between the two cameras based on effective field of view, resolution, and the retinal zone being imaged. Translational Relevance: The choice of which ultra-widefield camera to be used for screening DR can be based on the greater prevalence of central versus peripheral retinal lesions noted in the patient population seen in a clinical practice

Defining the relationship between Plasmodium falciparum parasite rate and clinical disease: statistical models for disease burden estimation

<p>Abstract</p> <p>Background</p> <p>Clinical malaria has proven an elusive burden to enumerate. Many cases go undetected by routine disease recording systems. Epidemiologists have, therefore, frequently defaulted to actively measuring malaria in population cohorts through time. Measuring the clinical incidence of malaria longitudinally is labour-intensive and impossible to undertake universally. There is a need, therefore, to define a relationship between clinical incidence and the easier and more commonly measured index of infection prevalence: the "parasite rate". This relationship can help provide an informed basis to define malaria burdens in areas where health statistics are inadequate.</p> <p>Methods</p> <p>Formal literature searches were conducted for <it>Plasmodium falciparum </it>malaria incidence surveys undertaken prospectively through active case detection at least every 14 days. The data were abstracted, standardized and geo-referenced. Incidence surveys were time-space matched with modelled estimates of infection prevalence derived from a larger database of parasite prevalence surveys and modelling procedures developed for a global malaria endemicity map. Several potential relationships between clinical incidence and infection prevalence were then specified in a non-parametric Gaussian process model with minimal, biologically informed, prior constraints. Bayesian inference was then used to choose between the candidate models.</p> <p>Results</p> <p>The suggested relationships with credible intervals are shown for the Africa and a combined America and Central and South East Asia regions. In both regions clinical incidence increased slowly and smoothly as a function of infection prevalence. In Africa, when infection prevalence exceeded 40%, clinical incidence reached a plateau of 500 cases per thousand of the population <it>per annum</it>. In the combined America and Central and South East Asia regions, this plateau was reached at 250 cases per thousand of the population <it>per annum</it>. A temporal volatility model was also incorporated to facilitate a closer description of the variance in the observed data.</p> <p>Conclusion</p> <p>It was possible to model a relationship between clinical incidence and <it>P. falciparum </it>infection prevalence but the best-fit models were very noisy reflecting the large variance within the observed opportunistic data sample. This continuous quantification allows for estimates of the clinical burden of <it>P. falciparum </it>of known confidence from wherever an estimate of <it>P. falciparum </it>prevalence is available.</p

Optical spectroscopy of Gaia detected protostars with DOT: can we probe protostellar photospheres?

Optical spectroscopy offers the most direct view of the stellar properties and the accretion indicators. Standard accretion tracers, such as $H\beta$, $H\alpha$, and, Ca II triplet lines, and most photospheric features, fall in the optical wavelengths. However, these tracers are not readily observable from deeply embedded protostars because of the large line of sight extinction (Av $\sim$ 50-100 mag) toward them. In some cases, however, it is possible to observe protostars at optical wavelengths if the outflow cavity is aligned along the line-of-sight that allows observations of the photosphere, or the envelope is very tenuous and thin such that the extinction is low. In such cases, we can not only detect these protostars at optical wavelengths but also follow up spectroscopically. We have used the HOPS catalog (Furlan et al. 2016) of protostars in Orion to search for optical counterparts for protostars in the Gaia DR3 survey. Out of the 330 protostars in the HOPS sample, an optical counterpart within 2" is detected for 62 of the protostars. For 17 out of 62 optically detected protostars, we obtained optical spectra { (between 5500 to 8900 $\AA$) using the Aries-Devasthal Faint Object Spectrograph \& Camera (ADFOSC) on the 3.6-m Devasthal Optical Telescope (DOT) and Hanle Faint Object Spectrograph Camera (HFOSC) on 2-m Himalayan Chandra Telescope (HCT)}. We detect strong photospheric features, such as the TiO bands in the spectra {(of 4 protostars)}, hinting that photospheres can form early on in the star formation process. We further determined the spectral types of protostars, which show photospheres similar to a late M-type. Mass accretion rates derived for the protostars are similar to those found for T-Tauri stars, in the range of 10$^{-7}$ to 10$^{-8}$ $M_\odot$/yr.Comment: 9 pages, 5 figures accepted in Journal of Astrophysics and Astronomy as part of the "Star formation studies in the context of NIR instruments on 3.6m DOT" special issu

Polymorphisms of TNF-enhancer and gene for FcγRIIa correlate with the severity of falciparum malaria in the ethnically diverse Indian population

<p>Abstract</p> <p>Background</p> <p>Susceptibility/resistance to <it>Plasmodium falciparum </it>malaria has been correlated with polymorphisms in more than 30 human genes with most association analyses having been carried out on patients from Africa and south-east Asia. The aim of this study was to examine the possible contribution of genetic variants in the <it>TNF </it>and <it>FCGR2A </it>genes in determining severity/resistance to <it>P. falciparum </it>malaria in Indian subjects.</p> <p>Methods</p> <p>Allelic frequency distribution in populations across India was first determined by typing genetic variants of the <it>TNF </it>enhancer and the <it>FCGR2A </it>G/A SNP in 1871 individuals from 55 populations. Genotyping was carried out by DNA sequencing, single base extension (SNaPshot), and DNA mass array (Sequenom). Plasma TNF was determined by ELISA. Comparison of datasets was carried out by Kruskal-Wallis and Mann-Whitney tests. Haplotypes and LD plots were generated by PHASE and Haploview, respectively. Odds ratio (OR) for risk assessment was calculated using EpiInfo™ version 3.4.</p> <p>Results</p> <p>A novel single nucleotide polymorphism (SNP) at position -76 was identified in the <it>TNF </it>enhancer along with other reported variants. Five <it>TNF </it>enhancer SNPs and the <it>FCGR2A </it>R131H (G/A) SNP were analyzed for association with severity of <it>P. falciparum </it>malaria in a malaria-endemic and a non-endemic region of India in a case-control study with ethnically-matched controls enrolled from both regions. <it>TNF </it>-1031C and -863A alleles as well as homozygotes for the TNF enhancer haplotype CACGG (-1031T>C, -863C>A, -857C>T, -308G>A, -238G>A) correlated with enhanced plasma TNF levels in both patients and controls. Significantly higher TNF levels were observed in patients with severe malaria. Minor alleles of -1031 and -863 SNPs were associated with increased susceptibility to severe malaria. The high-affinity IgG2 binding FcγRIIa AA (131H) genotype was significantly associated with protection from disease manifestation, with stronger association observed in the malaria non-endemic region. These results represent the first genetic analysis of the two immune regulatory molecules in the context of <it>P. falciparum </it>severity/resistance in the Indian population.</p> <p>Conclusion</p> <p>Association of specific <it>TNF </it>and <it>FCGR2A </it>SNPs with cytokine levels and disease severity/resistance was indicated in patients from areas with differential disease endemicity. The data emphasizes the need for addressing the contribution of human genetic factors in malaria in the context of disease epidemiology and population genetic substructure within India.</p

Enabling nanomaterial, nanofabrication and cellular technologies for nanoneuromedicines

Nanoparticulate delivery systems represent an area of particular promise for nanoneuromedicines. They possess significant potential for desperately needed therapies designed to combat a range of disorders associated with aging. As such, the field was selected as the focus for the 2014 meeting of the American Society for Nanomedicine. Regenerative, protective, immune modulatory, anti-microbial and anti-inflammatory products, or imaging agents are readily encapsulated in or conjugated to nanoparticles and as such facilitate the delivery of drug payloads to specific action sites across the blood-brain barrier. Diagnostic imaging serves to precisely monitor disease onset and progression while neural stem cell replacement can regenerate damaged tissue through control of stem cell fates. These, taken together, can improve disease burden and limit systemic toxicities. Such enabling technologies serve to protect the nervous system against a broad range of degenerative, traumatic, metabolic, infectious and immune disorders