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Authors: Miaomiao Zhang, Sabrina Beckmann, Haluk Ertan, Rocky Chau, Mike Manefield

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Phenazines are a large class of nitrogen-containing aromatic heterocyclic compounds, which are almost exclusively produced by bacteria from diverse genera including Pseudomonas and Streptomyces. Phenazine-1-carboxylic acid (PCA) as one of 'core' phenazines are converted from chorismic acid before modified to other phenazine derivatives in different cells. Phenazines have attracted enormous interests because of their multiple roles on biocontrol, bacterial interaction, biofilm formation and fitness of their producers. However, in spite of ecological importance, degradation as a part of phenazines’ fate only have extremely limited attention now. Here, to isolate PCA-degrading bacteria, 200 mg L-1 PCA was supplied as sole carbon, nitrogen and energy source in minimal mineral medium. Quantitative PCR and Reverse-transcript PCR were employed to study abundance and activity of functional gene MFORT 16269 in PCA degradation, respectively. Intermediates and products of PCA degradation were identified with LC-MS/MS. After enrichment and isolation, a PCA-degrading strain was selected from soil and was designated as Rhodanobacter sp. PCA2 based on full 16S rRNA sequencing. As determined by HPLC, strain PCA2 consumed 200 mg L-1 (836 µM) PCA at a rate of 17.4 µM h-1, accompanying with significant cells yield from 1.92 × 105 to 3.11 × 106 cells per mL. Strain PCA2 was capable of degrading other phenazines as well, including phenazine (4.27 µM h-1), pyocyanin (2.72 µM h-1), neutral red (1.30 µM h-1) and 1-hydroxyphenazine (0.55 µM h-1). Moreover, during the incubation, transcript copies of MFORT 16269 gene increased significantly from 2.13 × 106 to 8.82 × 107 copies mL-1, which was 2.77 times faster than that of the corresponding gene copy number (2.20 × 106 to 3.32 × 107 copies mL-1), indicating that MFORT 16269 gene was activated and played roles on PCA degradation. As analyzed by LC-MS/MS, decarboxylation from the ring structure was determined as the first step of PCA degradation, followed by cleavage of nitrogen-containing ring by dioxygenase which catalyzed phenazine to nitrosobenzene. Subsequently, phenylhydroxylamine was detected after incubation for two days and was then transferred to aniline and catechol. Additionally, genomic and proteomic analyses were also carried out for strain PCA2. Overall, the findings presented here showed that a newly isolated strain Rhodanobacter sp. PCA2 was capable of degrading phenazines through decarboxylation and cleavage of nitrogen-containing ring, during which MFORT 16269 gene was activated and played important roles.

Keywords: decarboxylation, MFORT16269 gene, phenazine-1-carboxylic acid degradation, Rhodanobacter sp. PCA2

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Authors: Mohammadjavad Sotoudeheian

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COVID-19, which began in December 2019, uses the angiotensin-converting enzyme 2 (ACE2) receptor to enter and spread through the cells. ACE2 mRNA is present in almost every organ, including nasopharynx, lung, as well as the brain. Ports of entry of SARS-CoV-2 into the central nervous system (CNS) may include arterial circulation, while viremia is remarkable. However, it is imperious to develop neurological symptoms evaluation CSF analysis in patients with COVID-19, but theoretically, ACE2 receptors are expressed in cerebellar cells and may be a target for SARS-CoV-2 infection in the brain. Recent evidence agrees that SARS-CoV-2 can impact the brain through direct and indirect injury. Two biomarkers for CNS injury, glial fibrillary acidic protein (GFAP) and neurofilament light chain (NFL) detected in the plasma of patients with COVID-19. NFL, an axonal protein expressed in neurons, is related to axonal neurodegeneration, and GFAP is over-expressed in CNS inflammation. GFAP cytoplasmic accumulation causes Schwan cells to misfunction, so affects myelin generation, reduces neuroskeletal support over NfLs during CNS inflammation, and leads to axonal degeneration. Interleukin-6 (IL-6), which extensively over-express due to interleukin storm during COVID-19 inflammation, regulates gene expression, as well as GFAP through STAT molecular pathway. IL-6 also impresses the phosphoinositide 3-kinase (PI3K)/STAT/smads pathway. The PI3K/ protein kinase B (Akt) pathway is the main modulator upstream of the mammalian target of rapamycin (mTOR), and alterations in this pathway are common in neurodegenerative diseases. Most neurodegenerative diseases show a disruption of autophagic function and display an abnormal increase in protein aggregation that promotes cellular death. Therefore, induction of autophagy has been recommended as a rational approach to help neurons clear abnormal protein aggregates and survive. The mTOR is a major regulator of the autophagic process and is regulated by cellular stressors. The mTORC1 pathway and mTORC2, as complementary and important elements in mTORC1 signaling, have become relevant in the regulation of the autophagic process and cellular survival through the extracellular signal-regulated kinase (ERK) pathway.

Keywords: mTORC1, COVID-19, PI3K, autophagy, neurodegeneration

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Authors: Marwa M. Abu-Serie, Marwa M. Eltarahony

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The search for reliable, effective, and safe nanoparticles (NPs) as a treatment for cancer is a pressing priority. In this study, Cu-NPs were fabricated by Streptomyces cyaneofuscatus through simultaneous bioreduction strategy of copper nitrate salt. The as-prepared Cu-NPs subjected to structural analysis; energy-dispersive X-ray spectroscopy, elemental mapping, X-ray diffraction, transmission electron microscopy, and ζ-potential. These biological synthesized Cu-NPs were mixed with disulfiram (DS), forming a nanocomplex of Cu-DS with a size of ~135 nm. The prepared nanocomplex (nanoCu-DS) exhibited higher anticancer activity than that of free complex of DS-Cu, Cu-NPs, and DS alone. This was illustrated by the lowest IC50 of nanoCu-DS (< 4 µM) against human breast and liver cancer cell lines comparing with DS-Cu, Cu-NPs, and DS (~8, 22.98-33.51 and 11.95-14.86, respectively). Moreover, flow cytometric analysis confirmed that higher apoptosis percentage range of nanoCu-DS-treated in MDA-MB 231, MCF-7, Huh-7, and HepG-2 cells (51.24-65.28%) than free complex of Cu-DS ( < 4.5%). Regarding inhibition potency of liver and breast cancer cell migration, no significant difference was recorded between free and nanocomplex. Furthermore, nanoCu-DS suppressed gene expression of β-catenine, Akt, and NF-κB and upregulated p53 expression (> 3, >15, > 5 and ≥ 3 folds, respectively) more efficiently than free complex (all ~ 1 fold) in MDA-MB 231 and Huh-7 cells. Our finding proved this prepared nano complex has a powerful anticancer activity relative to free complex, thereby offering a promising cancer treatment.

Keywords: biologically prepared Cu-NPs, breast cancer cell lines, liver cancer cell lines, nanoCu- disulfiram

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Authors: Eiji Nakamachi, Ryota Sakiyama, Koji Yamamoto, Yusuke Morita, Hidetoshi Sakamoto

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The regeneration of injured central nerve network caused by the cerebrovascular accidents is difficult, because of poor regeneration capability of central nerve system composed of the brain and the spinal cord. Recently, new regeneration methods such as transplant of nerve cells and supply of nerve nutritional factor were proposed and examined. However, there still remain many problems with the canceration of engrafted cells and so on and it is strongly required to establish an efficacious treating method of a central nerve system. Blackman proposed the electromagnetic stimulation method to enhance the axonal nerve extension. In this study, we try to design and fabricate a new three-dimensional (3D) bio-reactor, which can load a uniform AC magnetic field stimulation on PC12 cells in the extracellular environment for enhancement of an axonal nerve extension and 3D nerve network generation. Simultaneously, we measure the morphology of PC12 cell bodies, axons, and dendrites by the multiphoton excitation fluorescence microscope (MPM) and evaluate the effectiveness of the uniform AC magnetic stimulation to enhance the axonal nerve extension. Firstly, we designed and fabricated the uniform AC magnetic field stimulation bio-reactor. For the AC magnetic stimulation system, we used the laminated silicon steel sheets for a yoke structure of 3D chamber, which had a high magnetic permeability. Next, we adopted the pole piece structure and installed similar specification coils on both sides of the yoke. We searched an optimum pole piece structure using the magnetic field finite element (FE) analyses and the response surface methodology. We confirmed that the optimum 3D chamber structure showed a uniform magnetic flux density in the PC12 cell culture area by using FE analysis. Then, we fabricated the uniform AC magnetic field stimulation bio-reactor by adopting analytically determined specifications, such as the size of chamber and electromagnetic conditions. We confirmed that measurement results of magnetic field in the chamber showed a good agreement with FE results. Secondly, we fabricated a dish, which set inside the uniform AC magnetic field stimulation of bio-reactor. PC12 cells were disseminated with collagen gel and could be 3D cultured in the dish. The collagen gel were poured in the dish. The collagen gel, which had a disk shape of 6 mm diameter and 3mm height, was set on the membrane filter, which was located at 4 mm height from the bottom of dish. The disk was full filled with the culture medium inside the dish. Finally, we evaluated the effectiveness of the uniform AC magnetic field stimulation to enhance the nurve axonal extension. We confirmed that a 6.8 increase in the average axonal extension length of PC12 under the uniform AC magnetic field stimulation at 7 days culture in our bio-reactor, and a 24.7 increase in the maximum axonal extension length. Further, we confirmed that a 60 increase in the number of dendrites of PC12 under the uniform AC magnetic field stimulation. Finally, we confirm the availability of our uniform AC magnetic stimulation bio-reactor for the nerve axonal extension and the nerve network generation.

Keywords: nerve regeneration, axonal extension , PC12 cell, magnetic field, three-dimensional bio-reactor

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Authors: Noor Akhmazillah Fauzi, Mohammed Mehdi Farid, Filipa V. Silva

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The aim of this paper is to investigate if different concentration of Manuka honey (as a model food) has a major influence on the inactivation of Saccharomyces cerevisiae (as the testing microorganism) after subjecting it to HPP. Honey samples with different sugar concentrations (20, 30, 40, 50, 60 and 70 °Brix) were prepared aseptically using sterilized distilled water. No dilution of honey was made for the 80 °Brix sample. For the 0 °Brix sample (control), sterilized distilled water was used. Thermal treatment at 55 °C for 10 min (conventionally applied in honey pasteurisation in industry) was carried out for comparison purpose. S. cerevisiae cell numbers in honey samples were established before and after each HPP and thermal treatment. The number of surviving cells was determined after a proper dilution of the untreated and treated samples by the viable plate count method. S. cerevisiae cells, in different honey concentrations (0 to 80 °Brix), subjected to 600 MPa (at ambient temperature) showed an increasing resistance to inactivation with °Brix. A significant correlation (p < 0.05) between cell reduction and °Brix was found. Cell reduction in high pressure-treated samples varied linearly with °Brix (R2 > 0.9), confirming that the baroprotective effect of the food is due to sugar content. This study has practical implications in establishing efficient process design for commercial manufacturing of high sugar food products and on the potential use of HPP for such products.

Keywords: high pressure processing, honey, Saccharomyces cerevisiae, °Brix

Procedia PDF Downloads 353

Authors: Angelos Evangelou, Katerina Loizou, Loukas Koutsokeras, Orestes Marangos, Giorgos Constantinides, Stylianos Yiatros, Katerina Sofocleous, Vasileios Drakonakis

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Owing to their outstanding strength to weight properties, carbon fiber reinforced polymer (CFRPs) composites have attracted significant attention finding use in various fields (sports, automotive, transportation, etc.). The current momentum indicates that there is an increasing demand for their employment in high value bespoke applications such as avionics and electronic casings, damage sensing structures, EMI (electromagnetic interference) structures that dictate the use of materials with increased electrical conductivity both in-plane and through the thickness. Several efforts by research groups have focused on enhancing the through-thickness electrical conductivity of FRPs, in an attempt to combine the intrinsically high relative strengths exhibited with improved z-axis electrical response as well. However, only a limited number of studies deal with printing of nano-enhanced polymer inks to produce a pattern on dry fabric level that could be used to fabricate CFRPs with improved through thickness electrical conductivity. The present study investigates the employment of screen-printing process on technical dry fabrics using nano-reinforced polymer-based inks to achieve the required through thickness conductivity, opening new pathways for the application of fiber reinforced composites in niche products. Commercially available inks and in-house prepared inks reinforced with electrically conductive nanoparticles are employed, printed in different patterns. The aim of the present study is to investigate both the effect of the nanoparticle concentration as well as the droplet patterns (diameter, inter-droplet distance and coverage) to optimize printing for the desired level of conductivity enhancement in the lamina level. The electrical conductivity is measured initially at ink level to pinpoint the optimum concentrations to be employed using a “four-probe” configuration. Upon printing of the different patterns, the coverage of the dry fabric area is assessed along with the permeability of the resulting dry fabrics, in alignment with the fabrication of CFRPs that requires adequate wetting by the epoxy matrix. Results demonstrated increased electrical conductivities of the printed droplets, with increase of the conductivity from the benchmark value of 0.1 S/M to between 8 and 10 S/m. Printability of dense and dispersed patterns has exhibited promising results in terms of increasing the z-axis conductivity without inhibiting the penetration of the epoxy matrix at the processing stage of fiber reinforced composites. The high value and niche prospect of the resulting applications that can stem from CFRPs with increased through thickness electrical conductivities highlights the potential of the presented endeavor, signifying screen printing as the process to to nano-enable z-axis electrical conductivity in composite laminas. This work was co-funded by the European Regional Development Fund and the Republic of Cyprus through the Research and Innovation Foundation (Project: ENTERPRISES/0618/0013).

Keywords: CFRPs, conductivity, nano-reinforcement, screen-printing

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Authors: Kkochorong Park, Keun Cheon Kim, Hyoban Lee, Eun Ju Lee, Bongsoo Kim

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Introduction of biomaterials such as DNA, RNA, proteins is important for many research areas. There are many methods to introduce biomaterials into living organisms like tissue and cells. To introduce biomaterials, several indirect methods including virus‐mediated delivery, chemical reagent (i.e., lipofectamine), electrophoresis have been used. Such methods are passive delivery using an endocytosis process of cell, reducing an efficiency of delivery. Unlike the indirect delivery method, it has been reported that a direct delivery of exogenous biomolecules into nucleus have been more efficient to expression or integration of biomolecules. Nano-sized material is beneficial for detect signal from cell or deliver stimuli/materials into the cell at cellular and molecular levels, due to its similar physical scale. Especially, because 1 dimensional (1D) nanomaterials such as nanotube, nanorod and nanowire with high‐aspect ratio have nanoscale geometry and excellent mechanical, electrical, and chemical properties, they could play an important role in molecular and cellular biology. In this study, by using single crystalline 1D noble metal nanowire, we fabricated nano-sized 1D injector which can successfully interface with living cells and directly deliver biomolecules into several types of cell line (i.e., stem cell, mammalian embryo) without inducing detrimental damages on living cell. This nano-bio technology could be a promising and robust tool for introducing exogenous biomaterials into living organism.

Keywords: DNA, gene delivery, nanoinjector, nanowire

Procedia PDF Downloads 275

Authors: Melanie Ostermann, Eivind Birkeland, Ying Xue, Alexander Sauter, Mihaela R. Cimpan

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Background: New reliable and robust techniques to assess biological effects of nanomaterials (NMs) in vitro are needed to speed up safety analysis and to identify key physicochemical parameters of NMs, which are responsible for their acute cytotoxicity. The central aim of this study was to validate and evaluate the applicability and reliability of an impedance-based flow cytometry (IFC) technique for the high-throughput screening of NMs. Methods: Eight inorganic NMs from the European Commission Joint Research Centre Repository were used: NM-302 and NM-300k (Ag: 200 nm rods and 16.7 nm spheres, respectively), NM-200 and NM- 203 (SiO₂: 18.3 nm and 24.7 nm amorphous, respectively), NM-100 and NM-101 (TiO₂: 100 nm and 6 nm anatase, respectively), and NM-110 and NM-111 (ZnO: 147 nm and 141 nm, respectively). The aim was to assess the biological effects of these materials on human monoblastoid (U937) cells. Dispersions of NMs were prepared as described in the NANOGENOTOX dispersion protocol and cells were exposed to NMs at relevant concentrations (2, 10, 20, 50, and 100 µg/mL) for 24 hrs. The change in electrical impedance was measured at 0.5, 2, 6, and 12 MHz using the IFC AmphaZ30 (Amphasys AG, Switzerland). A traditional toxicity assay, Trypan Blue Dye Exclusion assay, and dark-field microscopy were used to validate the IFC method. Results: Spherical Ag particles (NM-300K) showed the highest toxic effect on U937 cells followed by ZnO (NM-111 ≥ NM-110) particles. Silica particles were moderate to non-toxic at all used concentrations under these conditions. A higher toxic effect was seen with smaller sized TiO2 particles (NM-101) compared to their larger analogues (NM-100). No interferences between the IFC and the used NMs were seen. Uptake and internalization of NMs were observed after 24 hours exposure, confirming actual NM-cell interactions. Conclusion: Results collected with the IFC demonstrate the applicability of this method for rapid nanotoxicity assessment, which proved to be less prone to nano-related interference issues compared to some traditional toxicity assays. Furthermore, this label-free and novel technique shows good potential for up-scaling in directions of an automated high-throughput screening and for future NM toxicity assessment. This work was supported by the EC FP7 NANoREG (Grant Agreement NMP4-LA-2013-310584), the Research Council of Norway, project NorNANoREG (239199/O70), the EuroNanoMed II 'GEMN' project (246672), and the UH-Nett Vest project.

Keywords: cytotoxicity, high-throughput, impedance, nanomaterials

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Authors: Ana C. Sousa, Beatriz C. Santos, Fátima N. Serralha

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The Tagus estuary is heavily affected by industrial and urban activities, making bioremediation studies crucial for environmental preservation. Fuel contamination in the area can arise from various anthropogenic sources, such as oil spills from shipping, fuel storage and transfer operations, and industrial discharges. These pollutants can cause severe harm to the ecosystem and the organisms, including humans, that inhabit it. Nonetheless, there are always natural organisms with the ability to resist these pollutants and transform them into non-toxic or harmless substances, which defines the process of bioremediation. Exploring the microbial communities existing in soil and their capacity to break down hydrocarbons has the potential to enhance the development of more efficient bioremediation approaches. The aim of this investigation was to explore the existence of hydrocarbonoclastic microorganisms in six locations within the Tagus estuary, three on the north bank: Trancão River, Praia Fluvial do Cais das Colinas and Praia de Algés, and three on the south bank: Praia Fluvial de Alcochete, Praia Fluvial de Alburrica, and Praia da Trafaria. In all studied locations, native microorganisms of the genus Pseudomonas were identified. The bioremediation rate of common hydrocarbons like gasoline, hexane, and toluene was assessed using the redox indicator 2,6-dichlorophenolindophenol (DCPIP). Effective hydrocarbon-degrading bacterial strains were identified in all analyzed areas, despite adverse environmental conditions. The highest bioremediation rates were achieved for gasoline (68%) in Alburrica, hexane (65%) in Algés, and toluene (79%) in Algés. Generally, the bacteria demonstrated efficient degradation of hydrocarbons added to the culture medium, with higher rates of aerobic biodegradation of hydrocarbons observed. These findings underscore the necessity for further in situ studies to better comprehend the relationship between native microbial communities and the potential for pollutant degradation in soil.

Keywords: biodegradability rate, hydrocarbonoclastic microorganisms, soil bioremediation, tagus estuary

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Authors: Ilya B. Tsyrlov, Dmitry Y. Oshchepkov

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A computational search for dioxin response elements (DREs) in genes of proteins comprising the Ah receptor (AhR) cytosolic core complex was performed by highly efficient tool SITECON. Eventually, the following number of new DREs in 5’flanking region was detected by SITECON: one in AHR gene, five in XAP2, eight in HSP90AA1, and three in HSP90AB1 genes. Numerous DREs found in genes of AhR and AhR cytosolic complex members would shed a light on potential mechanisms of expression, the stoichiometry of unliganded AhR core complex, and its degradation vs biosynthesis dynamics resulted from treatment of target cells with the AhR most potent ligand, 2,3,7,8-TCDD. With human viruses, reduced susceptibility to TCDD of geneencoding HIV-1 P247 was justified by the only potential DRE determined in gag gene encoding HIV-1 P24 protein, whereas the regulatory region of CMV genes encoding IE gp/UL37 has five potent DRE, 1.65 kb/UL36 – six DRE, pp65 and pp71 – each has seven DRE, and pp150 – ten DRE. Also, from six to eight DRE were determined with SITECON in the regulatory region of HSV-1 IE genes encoding tegument proteins, UL36 and UL37, and of UL19 gene encoding bindingglycoprotein C (gC). So, TCDD in the low picomolar range may activate in human cells AhR: Arnt transcription pathway that triggers CMV and HSV-1 reactivation by binding to numerous promoter DRE within immediate-early (IE) genes UL37 and UL36, thus committing virus to the lytic cycle.

Keywords: dioxin response elements, Ah receptor, AhR: Arnt transcription pathway, human and viral genes

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Authors: Anupalli Roja Rani, Pavithra Dasari

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Background: Among several, breast cancer has emerged as the most common female cancer in developing countries. It is the most common cause of cancer-related deaths worldwide among women. It is a molecularly and clinically heterogeneous disease. Moreover, it is a hormone–dependent tumor in which estrogens can regulate the growth of breast cells by binding with estrogen receptors (ERs). Moreover, the use of natural products in cancer therapeutics is due to their properties of biocompatibility and less toxicity. Plants are the vast reservoirs for various bioactive compounds. Coleus barbatus (Lamiaceae) contains anticancer properties against several cancer cell lines. Method: In the present study, an attempt is being made to enrich the knowledge of the anticancer activity of pure compounds extracted from Coleus barbatus (Andrew). On human breast cancer cell lines MCF-7. Here in, we are assessing the antiproliferative activity of Coleus barbatus (Andrew) plant extracts against MCF 7 and also evaluating their toxicity in normal human mammary cell lines such as Human Mammary Epithelial Cells (HMEC). The active fraction of plant extract was further purified with the help of Flash chromatography, Medium Pressure Liquid Chromatography (MPLC) and preparative High-Performance Liquid Chromatography (HPLC). The structure of pure compounds will be elucidated by using modern spectroscopic methods like Nuclear magnetic resonance (NMR), Electrospray Ionisation Mass Spectrometry (ESI-MS) methods. Later, the growth inhibition morphological assessment of cancer cells and cell cycle analysis of purified compounds were assessed using FACS. The growth and progression of signaling molecules HER2, GRP78 was studied by secretion assay using ELISA and expression analysis by flow cytometry. Result: Cytotoxic effect against MCF-7 with IC50 values were derived from dose response curves, using six concentrations of twofold serially diluted samples, by SOFTMax Pro software (Molecular device) and respectively Ellipticine and 0.5% DMSO were used as a positive and negative control. Conclusion: The present study shows the significance of various bioactive compounds extracted from Coleus barbatus (Andrew) root material. It acts as an anti-proliferative and shows cytotoxic effects on human breast cancer cell lines MCF7. The plant extracts play an important role pharmacologically. The whole plant has been used in traditional medicine for decades and the studies done have authenticated the practice. Earlier, as described, the plant has been used in the ayurveda and homeopathy medicine. However, more clinical and pathological studies must be conducted to investigate the unexploited potential of the plant. These studies will be very useful for drug designing in the future.

Keywords: coleus barbatus, HPLC, MPLC, NMR, MCF7, flash chromatograph, ESI-MS, FACS, ELISA.

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Authors: Deepak Kakde, Steve Howdle, Derek Irvine, Cameron Alexander

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The poor aqueous solubility is one of the major obstacles in the formulation development of many drugs. Around 70% of drugs are poorly soluble in aqueous media. In the last few decades, micelles have emerged as one of the major tools for solubilization of hydrophobic drugs. Micelles are nanosized structures (10-100nm) obtained by self-assembly of amphiphilic molecules into the water. The hydrophobic part of the micelle forms core which is surrounded by a hydrophilic outer shell called corona. These core-shell structures have been used as a drug delivery vehicle for many years. Although, the utility of micelles have been reduced due to the lack of sustainable materials. In the present study, a novel methoxy poly(ethylene glycol)-b-poly(ε-decalactone) (mPEG-b-PεDL) copolymer was synthesized by ring opening polymerization (ROP) of renewable ε-decalactone (ε-DL) monomers on methoxy poly(ethylene glycol) (mPEG) initiator using 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD) as a organocatalyst. All the reactions were conducted in bulk to avoid the use of toxic organic solvents. The copolymer was characterized by nuclear magnetic resonance spectroscopy (NMR), gel permeation chromatography (GPC) and differential scanning calorimetry (DSC).The mPEG-b-PεDL block copolymeric micelles containing indomethacin (IND) were prepared by nanoprecipitation method and evaluated as drug delivery vehicle. The size of the micelles was less than 40nm with narrow polydispersity pattern. TEM image showed uniform distribution of spherical micelles defined by clear surface boundary. The indomethacin loading was 7.4% for copolymer with molecular weight of 13000 and drug/polymer weight ratio of 4/50. The higher drug/polymer ratio decreased the drug loading. The drug release study in PBS (pH7.4) showed a sustained release of drug over a period of 24hr. In conclusion, we have developed a new sustainable polymeric material for IND delivery by combining the green synthetic approach with the use of renewable monomer for sustainable development of polymeric nanomedicine.

Keywords: dopolymer, ε-decalactone, indomethacin, micelles

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Authors: Pierre-Louis Lucas, Corinne Loutelier-Bourhis, Narimane Mati-Baouche, Philippe Chan Tchi-Song, Patrice Lerouge, Elodie Mathieu-Rivet, Muriel Bardor

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N-glycosylation is a post-translational modification taking place in the Endoplasmic Reticulum and the Golgi apparatus where defined glycan features are added on protein in a very specific sequence Asn-X-Thr/Ser/Cys were X can be any amino acid except proline. Because it is well-established that those N-glycans play a critical role in protein biological activity, protein half-life and that a different N-glycan structure may induce an immune response, they are very important in Biopharmaceuticals which are mainly glycoproteins bearing N-glycans. From now, most of the biopharmaceuticals are produced by mammalian cells like Chinese Hamster Ovary cells (CHO) for their N-glycosylation similar to the human, but due to the high production costs, several other species are investigated as the possible alternative system. In this purpose, the green microalgae Chlamydomonas reinhardtii was investigated as the potential production system for Biopharmaceuticals. This choice was influenced by the facts that C. reinhardtii is a well-study microalgae which is growing fast with a lot of molecular biology tools available. This organism is also producing N-glycan on its endogenous proteins. However, the analysis of the N-glycan structure of this microalgae has revealed some differences as compared to the human. Rather than in Human where the glycans are processed by key enzymes called N-acetylglucosaminyltransferase I and II (GnTI and GnTII) adding GlcNAc residue to form a GlcNAc₂Man₃GlcNAc₂ core N-glycan, C. reinhardtii lacks those two enzymes and possess a GnTI independent glycosylation pathway. Moreover, some enzymes like xylosyltransferases and methyltransferases not present in human are supposed to act on the glycans of C. reinhardtii. Furthermore, the recent structural study by mass spectrometry shows that the N-glycosylation precursor supposed to be conserved in almost all eukaryotic cells results in a linear Man₅GlcNAc₂ rather than a branched one in C. reinhardtii. In this work, we will discuss the new released MS information upon C. reinhardtii N-glycan structure and their impact on our attempt to modify the glycan in a Human manner. Two strategies will be discussed. The first one consisted in the study of Xylosyltransferase insertional mutants from the CLIP library in order to remove xyloses from the N-glycans. The second will go further in the humanization by transforming the microalgae with the exogenous gene from Toxoplasma gondii having an activity similar to GnTI and GnTII with the aim to synthesize GlcNAc₂Man₃GlcNAc₂ in C. reinhardtii.

Keywords: Chlamydomonas reinhardtii, N-glycosylation, glycosyltransferase, mass spectrometry, humanization

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Authors: Wei Xiao, Gangzhi Cai, Xingliang Qin, Hongyan Ren, Zaidong Hua, Zhe Zhu, Hongwei Xiao, Ximin Zheng, Jie Yao, Yanzhen Bi

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Myostatin (MSTN) is the master regulator of double muscling phenotype with overgrown muscle and decreased fatness in animals, but its action mode to regulate fat deposition remains to be elucidated. In this study a swin-specific pathway through which MSTN acts to regulate the fat deposition was deciphered. Deep sequenincing of the mRNA and miRNA of fat tissues of MSTN knockout (KO) and wildtype (WT) pigs discovered the positive correlation of myocyte enhancer factor 2C (MEF2C) and fat-inhibiting miR222 expression, and the inverse correlation of miR222 and stearoyl-CoA desaturase 5 (SCD5) expression. SCD5 is rodent-absent and expressed only in pig, sheep and cattle. Fatty acid spectrum of fat tissues revealed a lower percentage of oleoyl-CoA (18:1) and palmitoleyl CoA (16:1) in MSTN KO pigs, which are the catalyzing products of SCD5-mediated desaturation of steroyl CoA (18:0) and palmitoyl CoA (16:0). Blood metrics demonstrated a 45% decline of triglyceride (TG) content in MSTN KO pigs. In light of these observations we hypothesized that MSTN might act through MEF2C/miR222/SCD5 pathway to regulate desaturation of fatty acid as well as triglyceride synthesis in pigs. To this end, real-time PCR and Western blotting were carried out to detect the expression of the three genes stated above. These experiments showed that MEF2C expression was up-regulated by nearly 2-fold, miR222 up-regulated by nearly 3-fold and SCD5 down-regulated by nearly 50% in MSTN KO pigs. These data were consistent with the expression change in deep sequencing analysis. Dual luciferase reporter was then used to confirm the regulation of MEF2C upon the promoter of miR222. Ecotopic expression of MEF2C in preadipocyte cells enhanced miR222 expression by 3.48-fold. CHIP-PCR identified a putative binding site of MEF2C on -2077 to -2066 region of miR222 promoter. Electrophoretic mobility shift assay (EMSA) demonstrated the interaction of MEF2C and miR222 promoter in vitro. These data indicated that MEF2C transcriptionally regulates the expression of miR222. Next, the regulation of miR222 on SCD5 mRNA as well as its physiological consequences were examined. Dual luciferase reporter testing revealed the translational inhibition of miR222 upon the 3´ UTR (untranslated region) of SCD5 in preadipocyte cells. Transfection of miR222 mimics and inhibitors resulted in the down-regulation and up-regulation of SCD5 in preadipocyte cells respectively, consistent with the results from reporter testing. RNA interference of SCD5 in preadipocyte cells caused 26.2% reduction of TG, in agreement with the results of TG content in MSTN KO pigs. In summary, the results above supported the existence of a molecular pathway that MSTN signals through MEF2C/miR222/SCD5 to regulate the fat deposition in pigs. This swine-specific pathway offers potential molecular markers for the development and breeding of a new pig line with optimised fatty acid composition. This would benefit human health by decreasing the takeup of saturated fatty acid.

Keywords: fat deposition, MEF2C, miR222, myostatin, SCD5, pig

Procedia PDF Downloads 129

Authors: Indira K., Valsamma Joseph, I. S. Bright

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Introduction :Biofuels could replace fossil fuels and reduce our carbon footprint on the planet by technological advancements needed for sustainable and economic fuel production. Micro algae have proven to be a promising source to meet the current energy demand because of high lipid content and production of high biomass rapidly. Marine diatoms, which are key contributors in the biofuel sector and also play a significant role in primary productivity and ecology with high biodiversity and genetic and chemical diversity, are less well understood than other microalgae for producing hydrocarbons. Method :The marine diatom samples selected for hydrocarbon analysis were a total of eleven, out of which 9 samples were from the culture collection of NCAAH, and the remaining two of them were isolated by serial dilution method to get a pure culture from a mixed culture of microalgae obtained from the various cruise stations (350&357) FORV Sagar Sampada along the west coast of India. These diatoms were mass cultured in F/2 media, and the biomass harvested. The crude extract was obtained from the biomass by homogenising with n-hexane, and the hydrocarbons was further obtained by passing the crude extract through 500mg Bonna Agela SPE column and the quantitative analysis was done by GCHRMS analysis using HP-5 column and Helium gas was used as a carrier gas(1ml/min). The injector port temperature was 2400C, the detector temperature was 2500C, and the oven was initially kept at 600C for 1 minute and increased to 2200C at the rate of 60C per minute, and the analysis of a mixture of long chain hydrocarbons was done .Results:In the qualitative analysis done, the most potent hydrocarbon was found to be Psammodictyon Panduriforme (NCAAH-9) with a hydrocarbon mass of 37.27mg/g of the biomass and 2.1% of the total biomass 0f 1.395g and the other potent producer is Biddulphia(NCAAH 6) with hydrocarbon mass of 25.4mg/g of biomass and percentage of hydrocarbon is 1.03%. In the quantitative analysis by GCHRMS, the long chain hydrocarbons found in most of the marine diatoms were undecane, hexadecane, octadecane 3ethyl 5,2 ethyl butyl, Eicosane7hexyl, hexacosane, heptacosane, heneicosane, octadecane 3 methyl, triacontane. The exact mass of the long chain hydrocarbons in all the marine diatom samples was found to be Nonadecane 12C191H40, Tritriacontane,13-decyl-13-heptyl 12C501H102, Octadecane,3ethyl-5-(2-ethylbutyl 12C261H54, tetratetracontane 12C441H89, Eicosane, 7-hexyl 12C261H54. Conclusion:All the marine diatoms screened produced long chain hydrocarbons which can be used as diesel fuel with good cetane value example, hexadecane, undecane. All the long chain hydrocarbons can further undergo catalytic cracking to produce short chain alkanes which can give good octane values and can be used as gasoline. Optimisation of hydrocarbon production with the most potent marine diatom yielded long chain hydrocarbons of good fuel quality.

Keywords: biofuel, hydrocarbons, marine diatoms, screening

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Authors: Madhavi S. Apte, Milind Bhitre

Abstract:

In pharmaceutical research and new drug development, medicinal plants have important roles. Similarly, Indian dhak tree belonging to family Fabaceae has been widely used in the traditional Indian medical system of ‘Ayurveda’ for the treatment of a variety of ailments. Hence the cell viability study was undertaken to evaluate and compare the activity of extracts of various parts like flower, bark, leaf, seed by conducting MTT assay method along with other pharmacognostical studies. The methanolic extracts of bark, flowers, leaves, and seeds were used for the study. The cell viability MTT assay was performed using the standard operating procedures. The extracts were dissolved in DMSO and serially diluted with complete medium to get the concentrations range of test concentration. DMSO concentration was kept < 0.1% in all the samples. HUVEC cells maintained in appropriate conditions were seeded in 96 well plates and treated with different concentrations of the test samples and incubated at 37°C, 5% CO₂ for 96 hours. MTT reagent was added to the wells and incubated for 4 hours; the dark blue formazan product formed by the cells was dissolved in DMSO under a safety cabinet and read at 550nm. Percentage inhibitions were calculated and plotted with the concentrations used to calculate the IC50 values. The bark, flower, leaves and seed extracts have shown the cytotoxicity activity and can be further studied for antiangiogenesis activity.

Keywords: pharmacognosy, Cell viability, MTT assay, anti-angiogenesis

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Authors: Sophio Kobauri, Tengiz Kantaria, Temur Kantaria, David Tugushi, Nina Kulikova, Ramaz Katsarava

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Nanosized environmentally responsive materials are of special interest for various applications, including targeted drug to a considerable potential for treatment of many human diseases. The important technological advantages of nanoparticles (NPs) usage as drug carriers (nanocontainers) are their high stability, high carrier capacity, feasibility of encapsulation of both hydrophilic or hydrophobic substances, as well as a high variety of possible administration routes, including oral application and inhalation. NPs can also be designed to allow controlled (sustained) drug release from the matrix. These properties of NPs enable improvement of drug bioavailability and might allow drug dosage decrease. The targeted and controlled administration of drugs using NPs might also help to overcome drug resistance, which is one of the major obstacles in the control of epidemics. Various degradable and non-degradable polymers of both natural and synthetic origin have been used for NPs construction. One of the most promising for the design of NPs are amino acid-based biodegradable polymers (AABBPs) which can clear from the body after the fulfillment of their function. The AABBPs are composed of naturally occurring and non-toxic building blocks such as α-amino acids, fatty diols and dicarboxylic acids. The particles designed from these polymers are expected to have an improved bioavailability along with a high biocompatibility. The present work deals with a systematic study of the preparation of NPs by cost-effective polymer deposition/solvent displacement method using AABBPs. The influence of the nature and concentration of surfactants, concentration of organic phase (polymer solution), and the ratio organic phase/inorganic (water) phase, as well as of some other factors on the size of the fabricated NPs have been studied. It was established that depending on the used conditions the NPs size could be tuned within 40-330 nm. As the next step of this research an evaluation of biocompatibility and bioavailability of the synthesized NPs has been performed, using two stable human cell culture lines – HeLa and A549. This part of study is still in progress now.

Keywords: amino acids, biodegradable polymers, nanoparticles (NPs), non-toxic building blocks

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Authors: Kyriaki Hatziagapiou, Eleni Kakouri, Konstantinos Bethanis, Alexandra Nikola, Eleni Koniari, Charalabos Kanakis, Elias Christoforides, George Lambrou, Petros Tarantilis

Abstract:

Medulloblastoma is a highly invasive tumour, as it tends to disseminate throughout the central nervous system early in its course. Despite the high 5-year-survival rate, a significant number of patients demonstrate serious long- or short-term sequelae (e.g., myelosuppression, endocrine dysfunction, cardiotoxicity, neurological deficits and cognitive impairment) and higher mortality rates, unrelated to the initial malignancy itself but rather to the aggressive treatment. A strong rationale exists for the use of Crocus sativus L (saffron) and its bioactive constituents (crocin, crocetin, safranal) as pharmaceutical agents, as they exert significant health-promoting properties. Crocins are water soluble carotenoids. Unlike other carotenoids, crocins are highly water-soluble compounds, with relatively low toxicity as they are not stored in adipose and liver tissues. Crocins have attracted wide attention as promising anti-cancer agents, due to their antioxidant, anti-inflammatory, and immunomodulatory effects, interference with transduction pathways implicated in tumorigenesis, angiogenesis, and metastasis (disruption of mitotic spindle assembly, inhibition of DNA topoisomerases, cell-cycle arrest, apoptosis or cell differentiation) and sensitization of cancer cells to radiotherapy and chemotherapy. The current research aimed to study the potential cytotoxic effect of crocins on TE671 medulloblastoma cell line, which may be useful in the optimization of existing and development of new therapeutic strategies. Crocins were extracted from stigmas of saffron in ultrasonic bath, using petroleum-ether, diethylether and methanol 70%v/v as solvents and the final extract was lyophilized. Identification of crocins according to high-performance liquid chromatography (HPLC) analysis was determined comparing the UV-vis spectra and the retention time (tR) of the peaks with literature data. For the biological assays crocin was diluted to nuclease and protease free water. TE671 cells were incubated with a range of concentrations of crocins (16, 8, 4, 2, 1, 0.5 and 0.25 mg/ml) for 24, 48, 72 and 96 hours. Analysis of cell viability after incubation with crocins was performed with Alamar Blue viability assay. The active ingredient of Alamar Blue, resazurin, is a blue, nontoxic, cell permeable compound virtually nonfluorescent. Upon entering cells, resazurin is reduced to a pink and fluorescent molecule, resorufin. Viable cells continuously convert resazurin to resorufin, generating a quantitative measure of viability. The colour of resorufin was quantified by measuring the absorbance of the solution at 600 nm with a spectrophotometer. HPLC analysis indicated that the most abundant crocins in our extract were trans-crocin-4 and trans-crocin-3. Crocins exerted significant cytotoxicity in a dose and time-dependent manner (p < 0.005 for exposed cells to any concentration at 48, 72 and 96 hours versus cells not exposed); as their concentration and time of exposure increased, the reduction of resazurin to resofurin decreased, indicating reduction in cell viability. IC50 values for each time point were calculated ~3.738, 1.725, 0.878 and 0.7566 mg/ml at 24, 48, 72 and 96 hours, respectively. The results of our study could afford the basis of research regarding the use of natural carotenoids as anticancer agents and the shift to targeted therapy with higher efficacy and limited toxicity. Acknowledgements: The research was funded by Fellowships of Excellence for Postgraduate Studies IKY-Siemens Programme.

Keywords: crocetin, crocin, medulloblastoma, saffron

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Authors: Behnoush Dousti, Ye Choi, Gil S. Lee

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Microsupercapacitors (MScs) are known as the future of miniaturized energy sources that can be coupled to a battery to deliver stable and constant energy to microelectronics. Among all their counterparts, electrochemical microsupercapacitor have drawn the most research attention due to their higher power density and long cycle life. Designing the microstructure and choosing the electroactive materials are two significant factors that greatly affect the performance of the device. Here, we report successful fabrication and characterization of a microsupercapacitor with interdigitated structure based on Carbon nanotube sheets (CNT sheet). Novel structure of highly aligned CNT sheet as the electrode materials which also offers excellent conductivity and large surface area along with doping with nitrogen, enabled us to develop a device with serval order of magnitude higher electrochemical performance than the pristine CNT in aqueous electrolyte including high specific capacitance and rate capabilities and excellent cycle life over 10000 cycles. Geometric parameters such as finger width and gap size were also studied and it was shown the device performance is much depended on them. Results of this study confirms the potential of CNT sheet for future energy storage devices.

Keywords: carbon nanotube, energy storage systems, microsupercapacitor, nitrogen doping

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Authors: Tamunotonye Mayowa Braide, Isaac Oluwatayo

Abstract:

This study investigates multidimensional poverty, and its correlates among rural households in Sekhukhune and Capricorn District municipalities (SDM & CDM) in Limpopo Province, South Africa. Primary data were collected from 407 rural households selected through purposive and simple random sampling techniques. Analytical techniques employed include descriptive statistics, principal component analysis (PCA), and the Alkire Foster (A-F) methodology. The results of the descriptive statistics showed there are more females (66%) than males (34%) in rural areas of Limpopo Province, with about 45% of them having secondary school education as the highest educational level attained and only about 3% do not have formal education. In the analysis of deprivation, eight dimensions of deprivation, constructed from 21 variables, were identified using the PCA. These dimensions include type and condition of dwelling water and sanitation, educational attainment and income, type of fuel for cooking and heating, access to clothing and cell phone, assets and fuel for light, health condition, crowding, and child health. In identifying the poor with poverty cut-off (0.13) of all indicators, about 75.9% of the rural households are deprived in 25% of the total dimensions, with the adjusted headcount ratio (M0) being 0.19. Multidimensional poverty estimates showed higher estimates of poor rural households with 71%, compared to 29%, which fall below the income poverty line. The study conducted poverty decomposition, using sub-groups within the area by examining regions and household characteristics. In SDM, there are more multidimensionally poor households than in CDM. The water and sanitation dimension is the largest contributor to the multidimensional poverty index (MPI) in rural areas of Limpopo Province. The findings can, therefore, assist in better design of welfare policy and target poverty alleviation programs and as well help in efficient resource allocation at the provincial and local municipality levels.

Keywords: Alkire-Foster methodology, Limpopo province, multidimensional poverty, principal component analysis, South Africa

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Authors: Shilpa Khobragade, Carlos Da Silva Granja, Niklas Sandström, Igor Efimov, Victor P. Ostanin, Wouter van der Wijngaart, David Klenerman, Sourav K. Ghosh

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Rapid, label-free and direct detection of pathogenic bacteria is critical for the prevention of disease outbreaks. This paper for the first time attempts to probe the nonlinear acoustic response of quartz crystal resonator (QCR) functionalized with specific DNA aptamers for direct detection and quantification of viable E. coli KCTC 2571 bacteria. DNA aptamers were immobilized through biotin and streptavidin conjugation, onto the gold surface of QCR to capture the target bacteria and the detection was accomplished by shift in amplitude of the peak 3f signal (3 times the drive frequency) upon binding, when driven near fundamental resonance frequency. The developed nonlinear acoustic aptasensor system demonstrated better reliability than conventional resonance frequency shift and energy dissipation monitoring that were recorded simultaneously. This sensing system could directly detect 10⁽⁵⁾ cells/mL target bacteria within 30 min or less and had high specificity towards E. coli KCTC 2571 bacteria as compared to the same concentration of S.typhi bacteria. Aptasensor response was observed for the bacterial suspensions ranging from 10⁽⁵⁾-10⁽⁸⁾ cells/mL. Conclusively, this nonlinear acoustic aptasensor is simple to use, gives real-time output, cost-effective and has the potential for rapid, specific, label-free direction detection of bacteria.

Keywords: acoustic, aptasensor, detection, nonlinear

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Authors: Sambit Supriya Dash, Rahul Ravi R, Vikram Ramanan, Vinayak Malhotra

Abstract:

Combustion in itself is a complex phenomenon that involves the interaction and interplay of multiple phenomena, the combined effect of which gives rise to the common flame that we see and use in our daily life applications from cooking to propelling our vehicles to space. The most important thing that goes unnoticed about these flames is the effect of the various phenomena from its surrounding environment that affects its behavior and properties. These phenomena cause a variety of energy interactions that lead to various types of energy transformations which in turn affect the flame behavior. This paper focuses on experimentally investigating the effect of one such phenomenon, which is the acoustics or sound energy on diffusion flames. The subject in itself is extensively studied upon as thermo-acoustics globally, whereas the current work focuses on studying its effect on soot formation on diffusion flames. The said effect is studied in this research work by the use of a butane as fuel, fitted with a nozzle that houses 3 arrays consisting of 4 holes each that are placed equidistant to each other and the resulting flame impinged with sound from two independent and similar sound sources that are placed equidistant from the centre of the flame. The entire process is systematically video graphed using a 60 fps regular CCD and analysed for variation in flame heights and flickering frequencies where the fuel mass flow rate is maintained constant and the configuration of entrainment holes and frequency of sound are varied, whilst maintaining constant ambient atmospheric conditions. The current work establishes significant outcomes on the effect of acoustics on soot formation; it is noteworthy that soot formation is the main cause of pollution and a major cause of inefficiency of current propulsion systems. This work is one of its kinds, and its outcomes are widely applicable to commercial and domestic appliances that utilize combustion for energy generation or propulsion and help us understand them better, so that we can increase their efficiency and decrease pollution.

Keywords: thermoacoustics, entrainment, propulsion system, efficiency, pollution

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Authors: P. Selyshchev

Abstract:

Construction materials for nuclear facilities are operated under extreme thermal and radiation conditions. First of all, they are nuclear fuel, fuel assemblies, and reactor vessel. It places high demands on the control of their state, stability of their state, and their operating conditions. An irradiated material is a typical example of an open non-equilibrium system with nonlinear feedbacks between its elements. Fluxes of energy, matter and entropy maintain states which are far away from thermal equilibrium. The links that arise under irradiation are inherently nonlinear. They form the mechanisms of feed-backs that can lead to instability. Due to this instability the temperature of the sample, heat transfer, and the defect density can exceed the steady-state value in several times. This can lead to change of typical operation and an accident. Therefore, it is necessary to take into account the thermal instability to avoid the emergency situation. The point is that non-thermal energy can be accumulated in materials because irradiation produces defects (first of all these are vacancies and interstitial atoms), which are metastable. The stored energy is about energy of defect formation. Thus, an annealing of the defects is accompanied by releasing of non-thermal stored energy into thermal one. Temperature of the material grows. Increase of temperature results in acceleration of defect annealing. Density of the defects drops and temperature grows more and more quickly. The positive feed-back is formed and self-reinforcing annealing of radiation defects develops. To describe these phenomena a theoretical approach to thermal instability is developed via formalism of complex systems. We consider system of nonlinear differential equations for different components of microstructure and temperature. The qualitative analysis of this non-linear dynamical system is carried out. Conditions for development of instability have been obtained. Points of bifurcation have been found. Convenient way to represent obtained results is a set of phase portraits. It has been shown that different regimes of material state under irradiation can develop. Thus degradation of irradiated material can be limited by means of choice appropriate kind of evolution of materials under irradiation.

Keywords: irradiation, material, non-equilibrium state, nonlinear feed-back, thermal instability

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Authors: Juliana Shimara Pires Ferrão, Letícia Palmeira Pinto, Francisco Palma Rennó, Francisco Javier Hernandez Blazquez

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The ruminant stomach is a complex and multi-chambered organ. Although the true stomach (abomasum) is fully differentiated and functional at birth, the same does not occur with the rumen chamber. At this moment, rumen papillae are small or nonexistent. The papillae only fully develop after weaning and during calf growth. Papillae development and ruminal epithelium specialization during the fetus growth and at birth must be two interdependent processes that will prepare the rumen to adapt to ruminant adult feeding. The microscopic study of rumen epithelium at these early phases of life is important to understand how this structure prepares the rumen to deal with the following weaning processes and its functional activation. Samples of ruminal mucosa of bovine fetuses (110- and 150 day-old) and newborn calves were collected (dorsal and ventral portions) and processed for light and electron microscopy and immunohistochemistry. The basal cell layer of the stratified pavimentous epithelium present in different ruminal portions of the fetuses was thicker than the same portions of newborn calves. The superficial and intermediate epithelial layers of 150 day-old fetuses were thicker than those found in the other 2 studied ages. At this age (150 days), dermal papillae begin to invade the intermediate epithelial layer which gradually disappears in newborn calves. At birth, the ruminal papillae project from the epithelial surface, probably by regression of the epithelial cells (transitory cells) surrounding the dermal papillae. The PCNA cell proliferation index (%) was calculated for all epithelial samples. Fetuses 150 day-old showed increased cell proliferation in basal cell layer (Dorsal Portion: 84.2%; Ventral Portion: 89.8%) compared to other ages studied. Newborn calves showed an intermediate index (Dorsal Portion: 65.1%; Ventral Portion: 48.9%), whereas 110 day-old fetuses had the lowest proliferation index (Dorsal Portion: 57.2%; Ventral Portion: 20.6%). Regarding the transitory epithelium, 110 day-old fetuses showed the lowest proliferation index (Dorsal Portion: 44.6%; Ventral Portion: 20.1%), 150 day-old fetuses showed an intermediate proliferation index (Dorsal Portion: 57.5%; Ventral Portion: 71.1%) and newborn calves presented a higher proliferation index (Dorsal Portion: 75.1%; Ventral Portion: 19.6%). Under TEM, the 110- and 150 day-old fetuses presented thicker and poorly organized basal cell layer, with large nuclei and dense cytoplasm. In newborn calves, the basal cell layer was more organized and with fewer layers, but typically similar in both regions of the rumen. For the transitory epithelium, fetuses displayed larger cells than those found in newborn calves with less electrondense cytoplasm than that found in the basal cells. The ruminal dorsal portion has an overall higher cell proliferation rate than the ventral portion. Thus we can infer that the dorsal portion may have a higher cell activity than the ventral portion during ruminal development. Moreover, the basal cell layer is thicker in the 110- and 150 day-old fetuses than in the newborn calves. The transitory epithelium, which is much reduced, at birth may have a structural support function of the developing dermal papillae. When it regresses or is sheared off, the papillae are “carved out” from the surrounding epithelial layer.

Keywords: bovine, calf, epithelium, fetus, hematoxylin-eosin, immunohistochemistry, TEM, Rumen

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Authors: D. Suman, M. Venkateshwara Rao

Abstract:

Cancer is a disease caused by an uncontrolled division of abnormal cells in a part of the body, which is affecting millions of people leading to death. Even though there have been tremendous developments taken place over the last few decades the effective therapy for cancer is still not a reality. The existing techniques of cancer therapy are chemotherapy and radio therapy which are having their limitations in terms of the side effects, patient discomfort, radiation hazards and the localization of treatment. This paper describes a novel method for cancer therapy by using RF-hyperthermia application of nanoparticles. We have synthesized ferromagnetic nanoparticles and characterized by using XRD and TEM. These nanoparticles after the biocompatibility studies will be injected in to the body with a suitable tracer element having affinity to the specific tumor site. When RF energy is applied to the nanoparticles at the tumor site it produces heat of excess room temperature and nearly 41-45°C is sufficient to kill the tumor cells. We have designed a RF source generator provided with a temperature feedback controller to control the radiation induced temperature of the tumor site. The temperature control is achieved through a negative feedback mechanism of the thermocouple and a relay connected to the power source of the RF generator. This method has advantages in terms of its effect like localized therapy, less radiation, and no side effects. It has several challenges in designing the RF source provided with coils suitable for the tumour site, biocompatibility of the nanomaterials, cooling system design for the RF coil. If we can overcome these challenges this method will be a huge benefit for the society.

Keywords: hyperthermia, cancer therapy, RF source generator, nanoparticles

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Authors: Aulanni’am Aulanni’am

Abstract:

Incidence of Diabetes Mellitus (DM) progressively increasing it became a serious problem in Indonesia and it is a disease that government is priority to be addressed. The longer a person is suffering from diabetes the more likely to develop complications particularly diabetic patients who are not well maintained. Therefore, Incidence of Diabetes Mellitus needs to be done in the early diagnosis of pre-phase of the disease. In this pre-phase disease, already happening destruction of pancreatic beta cells and declining in beta cell function and the sign autoimmunity reactions associated with beta cell destruction. Type 1 DM is a multifactorial disease triggered by genetic and environmental factors, which leads to the destruction of pancreatic beta cells. Early marker of "beta cell autoreactivity" is the synthesis of autoantibodies against 65-kDa protein, which can be a molecule that can be detected early in the disease pathomechanism. The importance of early diagnosis of diabetic patients held in the phase of pre-disease is to determine the progression towards the onset of pancreatic beta cell destruction and take precautions. However, the price for this examination is very expensive ($ 150/ test), the anti-GAD65 abs examination cannot be carried out routinely in most or even in all laboratories in Indonesia. Therefore, production-based Rapid Test Recombinant Human Protein GAD65 with "Reverse Flow Immunchromatography Technique" in Indonesia is believed to reduce costs and improve the quality of care of patients with diabetes in Indonesia. Rapid Test Product innovation is very simple and suitable for screening and routine inspection of GAD65 autoantibodies. In the blood serum of patients with diabetes caused by autoimmunity, autoantibody-GAD65 is a major serologic marker to detect autoimmune reaction because their concentration level of stability.GAD65 autoantibodies can be found 10 years before clinical symptoms of diabetes. Early diagnosis is more focused to detect the presence autontibodi-GAD65 given specification and high sensitivity. Autoantibodies- GAD65 that circulates in the blood is a major indicator of the destruction of the islet cells of the pancreas. Results of research in collaboration with Biofarma has produced GAD65 autoantibodies based Rapid Test had conducted the soft launch of products and has been tested with the results of a sensitivity of 100 percent and a specificity between 90 and 96% compared with the gold standard (import product) which worked based on ELISA method.

Keywords: diabetes mellitus, GAD65 autoantibodies, rapid test, sensitivity, specificity

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Authors: Mehrnaz Mostafavi

Abstract:

Perovskite nanocrystals and quantum dots have emerged as leaders in the field of photovoltaic technologies, demonstrating exceptional light-harvesting abilities and stability. This study investigates the substantial progress and potential of these nano-sized materials in transforming solar energy conversion. The research delves into the foundational characteristics and production methods of perovskite nanocrystals and quantum dots, elucidating their distinct optical and electronic properties that render them well-suited for photovoltaic applications. Specifically, it examines their outstanding light absorption capabilities, enabling more effective utilization of a wider solar spectrum compared to traditional silicon-based solar cells. Furthermore, this paper explores the improved durability achieved in perovskite nanocrystals and quantum dots, overcoming previous challenges related to degradation and inconsistent performance. Recent advancements in material engineering and techniques for surface passivation have significantly contributed to enhancing the long-term stability of these nanomaterials, making them more commercially feasible for solar cell usage. The study also delves into the advancements in device designs that incorporate perovskite nanocrystals and quantum dots. Innovative strategies, such as tandem solar cells and hybrid structures integrating these nanomaterials with conventional photovoltaic technologies, are discussed. These approaches highlight synergistic effects that boost efficiency and performance. Additionally, this paper addresses ongoing challenges and research endeavors aimed at further improving the efficiency, stability, and scalability of perovskite nanocrystals and quantum dots in photovoltaics. Efforts to mitigate concerns related to material degradation, toxicity, and large-scale production are actively pursued, paving the way for broader commercial application. In conclusion, this paper emphasizes the significant role played by perovskite nanocrystals and quantum dots in advancing photovoltaic technologies. Their exceptional light-harvesting capabilities, combined with increased stability, promise a bright future for next-generation solar cells, ushering in an era of highly efficient and cost-effective solar energy conversion systems.

Keywords: perovskite nanocrystals, quantum dots, photovoltaic technologies, light-harvesting, solar energy conversion, stability, device designs

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Authors: Vivek Thorat, Suhasini Desai

Abstract:

The hybrid electric vehicle is widely accepted as a promising short to mid-term technical solution due to noticeably improved efficiency and low emissions at competitive costs. Retro fitment of hybrid components into a conventional vehicle for achieving better performance is the best solution so far. But retro fitment includes major modifications into a conventional vehicle with a high cost. This paper focuses on the development of a P3x hybrid prototype with rear wheel drive parallel hybrid electric Light Commercial Vehicle (LCV) with minimum and low-cost modifications. This diesel Hybrid LCV is different from another hybrid with regard to the powertrain. The additional powertrain consists of continuous contact helical gear pair followed by chain and sprocket as a coupler for traction motor. Vehicle powertrain which is designed for the intended high-speed application. This work focuses on targeting of design, development, and packaging of this unique parallel diesel-electric vehicle which is based on multimode hybrid advantages. To demonstrate the practical applicability of this transmission with P3x hybrid configuration, one concept prototype vehicle has been build integrating the transmission. The hybrid system makes it easy to retrofit existing vehicle because the changes required into the vehicle chassis are a minimum. The additional system is designed for mainly five modes of operations which are engine only mode, electric-only mode, hybrid power mode, engine charging battery mode and regenerative braking mode. Its driving performance, fuel economy and emissions are measured and results are analyzed over a given drive cycle. Finally, the output results which are achieved by the first vehicle prototype during experimental testing is carried out on a chassis dynamometer using MIDC driving cycle. The results showed that the prototype hybrid vehicle is about 27% faster than the equivalent conventional vehicle. The fuel economy is increased by 20-25% approximately compared to the conventional powertrain.

Keywords: P3x configuration, LCV, hybrid electric vehicle, ROMAX, transmission

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Authors: Tomokazu Shirai, Akihiko Kondo

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Technologies of metabolic engineering or synthetic biology are essential for effective microbial bioproduction. It is especially important to develop an in silico tool for designing a metabolic pathway producing an unnatural and valuable chemical such as fossil materials of fuel or plastics. We here demonstrated two in silico tools for designing novel metabolic pathways: BioProV and HyMeP. Furthermore, we succeeded in creating an artificial metabolic pathway by enzyme engineering.

Keywords: bioinformatics, metabolic engineering, synthetic biology, genome scale model

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Authors: D. Olszewska, J. Niewiedzial

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In some types of Li-ion batteries carbon in the form of graphite is used. Unfortunately, carbon materials, in particular graphite, have very good electrochemical properties, but increase their volume during charge/discharge cycles, which may even lead to an explosion of the cell. The cell element may be replaced by a composite material consisting of lithium-titanium oxide Li4Ti5O12 (LTO) modified with copper and nickel ions and carbon derived from sucrose. This way you can improve the conductivity of the material. LTO is appropriate only for applications which do not require high energy density because of its high operating voltage (ca. 1.5 V vs. Li/Li+). Specific capacity of Li4Ti5O12 is high enough for utilization in Li-ion batteries (theoretical capacity 175 mAh·g-1) but it is lower than capacity of graphite anodes. Materials based on Li4Ti5O12 do not change their volume during charging/discharging cycles, however, LTO has low conductivity. Another positive aspect of the use of sucrose in the carbon composite material is to eliminate the addition of carbon black from the anode of the battery. Therefore, the proposed materials contribute significantly to environmental protection and safety of selected lithium cells. New anode materials in order to obtain Li3.8Cu0.1Ni0.1Ti5O12 have been prepared by solid state synthesis using three-way: i) stoichiometric composition of Li2CO3, TiO2, CuO, NiO (A- Li3.8Cu0.1Ni0.1Ti5O12); ii) stoichiometric composition of Li2CO3, TiO2, Cu(NO3)2, Ni(NO3)2 (B-Li3.8Cu0.1Ni0.1Ti5O12); and iii) stoichiometric composition of Li2CO3, TiO2, CuO, NiO calcined with 10% of saccharose (Li3.8Cu0.1Ni0.1Ti5O12-C). Structure of materials was studied by X-ray diffraction (XRD). The electrochemical properties were performed using appropriately prepared cell Li|Li+|Li3.8Cu0.1Ni0.1Ti5O12 for cyclic voltammetry and discharge/charge measurements. The cells were periodically charged and discharged in the voltage range from 1.3 to 2.0 V applying constant charge/discharge current in order to determine the specific capacity of each electrode. Measurements at various values of the charge/discharge current (from C/10 to 5C) were carried out. Cyclic voltammetry investigation was carried out by applying to the cells a voltage linearly changing over time at a rate of 0.1 mV·s-1 (in the range from 2.0 to 1.3 V and from 1.3 to 2.0 V). The XRD method analyzes show that composite powders were obtained containing, in addition to the main phase, 4.78% and 4% TiO2 in A-Li3.8Cu0.1Ni0.1O12 and B-Li3.8Cu0.1Ni0.1O12, respectively. However, Li3.8Cu0.1Ni0.1O12-C material is three-phase: 63.84% of the main phase, 17.49 TiO2 and 18.67 Li2TiO3. Voltammograms of electrodes containing materials A-Li3.8Cu0.1Ni0.1O12 and B-Li3.8Cu0.1Ni0.1O12 are correct and repeatable. Peak cathode occurs for both samples at a potential approx. 1.52±0.01 V relative to a lithium electrode, while the anodic peak at potential approx. 1.65±0.05 V relative to a lithium electrode. Voltammogram of Li3.8Cu0.1Ni0.1Ti5O12-C (especially for the first measurement cycle) is not correct. There are large variations in values of specific current, which are not characteristic for materials LTO. From the point of view of safety and environmentally friendly production of Li-ion cells eliminating soot and applying Li3.8Cu0.1Ni0.1Ti5O12-C as an active material of an anode in lithium-ion batteries seems to be a good alternative to currently used materials.

Keywords: anode, Li-ion batteries, Li₄O₅O₁₂, spinel

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