Search results for: hemtopoietic stem cell transplantation

Authors: Yun-An Chen, Hung-Jun Lin, Tai-Horng Young, Der-Zen Liu

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Decellularized brain extracellular matrix had been shown that it has the ability to influence on cell proliferation, differentiation and associated cell phenotype. However, this scaffold is thought to have poor mechanical properties and rapid degradation, it is hard for cell recellularization. In this study, we used decellularized brain extracellular matrix combined with chitosan, which is naturally occurring polysaccharide and non-cytotoxic polymer, forming a 3-D scaffold for neural stem/precursor cells (NSPCs) regeneration. HE staining and DAPI fluorescence staining confirmed decellularized process could effectively vanish the cellular components from the brain. GAGs and collagen I, collagen IV were be showed a great preservation by Alcain staining and immunofluorescence staining respectively. Decellularized brain extracellular matrix was well mixed in chitosan to form a 3-D scaffold (DB-C scaffold). The pore size was approximately 50±10 μm examined by SEM images. Alamar blue results demonstrated NSPCs had great proliferation ability in DB-C scaffold. NSPCs that were cultured in this complex scaffold differentiated into neurons and astrocytes, as reveled by NSPCs expression of microtubule-associated protein 2 (MAP2) and glial fibrillary acidic protein (GFAP). In conclusion, DB-C scaffold may provide bioinformatics cues for NSPCs generation and aid for CNS injury functional recovery applications.

Keywords: brain, decellularization, chitosan, scaffold, neural stem/precursor cells

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Authors: Sepideh Hassanpour Khodaei

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Background/Objectives: The purpose of this study is to evaluate the effect of mummy substances on two issues of proliferation and production of matrix protein synthesis in wound healing. Methods: The methodology used for this aim involves isolating mesenchymal stem cells and human fibroblasts procured at Pastor Institute, Iran. The cells were treated with mummy substances separately and co-cultured between ASCs and WJSCs, and fibroblasts. Proliferation was assessed by Ki67 method in monolayer conditions. Synthesis of components of extracellular matrix (ECM) such as collagen type I, type III, and fibronectin 1 (FN1) was determined by qPCR. Results: The effects of adipocyte stem cells (ASCs), Wharton Jelly Stem Cells (WJSCs), and Mummy material on fibroblast proliferation and migration were evaluated. The present finding underlined the importance of Mummy material, ASCs, and WJSCs in the proliferation and migration of fibroblast cells. Furthermore, the expression of collagen I, III, and FN1 was increased in the presence of the above material and cells. Conclusion: This study presented an effective in vitro method for the healing process. Hence, the prospect of utilizing Mummy material and stem cell-based therapies in wound healing as a therapeutic approach is promising.

Keywords: mummy material, wound healing, adipose tissue, Wharton’s jelly

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Authors: Jimmy Jiun-Ming Su, Yuan-Min Lin

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Bioprinting has been applied to produce 3D cellular constructs for tissue engineering. Microextrusion printing is the most common used method. However, printing low viscosity bioink is a challenge for this method. Herein, we developed a new 3D printing system to fabricate cell-laden hydrogels via a DLP-based projector. The bioprinter is assembled from affordable equipment including a stepper motor, screw, LED-based DLP projector, open source computer hardware and software. The system can use low viscosity and photo-polymerized bioink to fabricate 3D tissue mimics in a layer-by-layer manner. In this study, we used gelatin methylacrylate (GelMA) as bioink for stem cell encapsulation. In order to reinforce the printed construct, surface modified hydroxyapatite has been added in the bioink. We demonstrated the silanization of hydroxyapatite could improve the crosslinking between the interface of hydroxyapatite and GelMA. The results showed that the incorporation of silanized hydroxyapatite into the bioink had an enhancing effect on the mechanical properties of printed hydrogel, in addition, the hydrogel had low cytotoxicity and promoted the differentiation of embedded human bone marrow stem cells (hBMSCs) and retinal pigment epithelium (RPE) cells. Moreover, this bioprinting system has the ability to generate microchannels inside the engineered tissues to facilitate diffusion of nutrients. We believe this 3D bioprinting system has potential to fabricate various tissues for clinical applications and regenerative medicine in the future.

Keywords: bioprinting, cell encapsulation, digital light processing, GelMA hydrogel

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Authors: Mekni Sabrine, Nouira Mariem

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Background and aim: Allogeneic hematopoietic stem cell transplantation is a curative treatment for several hematological diseases; however, it has a non-negligible morbidity and mortality depending on several prognostic factors, including pre-transplant hyperferritinemia. The aim of our study was to estimate the impact of hyperferritinemia on survivals and on the occurrence of post-transplant complications. Methods: It was a longitudinal study conducted over 8 years and including all patients who had a first allograft. The impact of pretransplant hyperferritinemia (ferritinemia ≥1500) on survivals was studied using the Kaplan Meier method and the COX model for uni- and multivariate analysis. The Khi-deux test and binary logistic regression were used to study the association between pretransplant ferritinemia and post-transplant complications. Results: One hundred forty patients were included with an average age of 26.6 years and a sex ratio (M/F)=1.4. Hyperferritinemia was found in 33% of patients. It had no significant impact on either overall survival (p=0.9) or event -free survival (p=0.6). In multivariate analysis, only the type of disease was independently associated with overall survival (p=0.04) and event-free survival (p=0.002). For post-allograft complications: The occurrence of early documented infections was independently associated with pretransplant hyperferritinemia (p=0.02) and the presence of acute graft versus host disease( GVHD) (p<10-3). The occurrence of acute GVHD was associated with early documented infection (p=0.002) and Cytomegalovirus reactivation (p<10-3). The occurrence of chronic GVHD was associated with the presence of Cytomegalovirus reactivation (p=0.006) and graft source (p=0.009). Conclusion: Our study showed the significant impact of pre-transplant hyperferritinemia on the occurrence of early infections but not on survivals. Early and more accurate assessment iron overload by other tests such as liver magnetic resonance imaging with initiation of chelating treatment could prevent the occurrence of such complications after transplantation.

Keywords: allogeneic, transplants, ferritin, survival

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Authors: Rafia S. Al-lamki, Jun Wang, Simon Pacey, Jordan Pober, John R. Bradley

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Tumor necrosis factor alpha (TNF), signaling through TNFR2, may act an autocrine growth factor for renal tubular epithelial cells. Clear cell renal carcinomas (ccRCC) contain cancer stem cells (CSCs) that give rise to progeny which form the bulk of the tumor. CSCs are rarely in cell cycle and, as non-proliferating cells, resist most chemotherapeutic agents. Thus, recurrence after chemotherapy may result from the survival of CSCs. Therapeutic targeting of both CSCs and the more differentiated bulk tumor populations may provide a more effective strategy for treatment of RCC. In this study, we hypothesized that TNFR2 signaling will induce CSCs in ccRCC to enter cell cycle so that treatment with ligands that engage TNFR2 will render CSCs susceptible to chemotherapy. To test this hypothesis, we have utilized wild-type TNF (wtTNF) or specific muteins selective for TNFR1 (R1TNF) or TNFR2 (R2TNF) to treat either short-term organ cultures of ccRCC and adjacent normal kidney (NK) tissue or cultures of CD133+ cells isolated from ccRCC and adjacent NK, hereafter referred to as stem cell-like cells (SCLCs). The effect of cyclophosphamide (CP), currently an effective anticancer agent, was tested on CD133+SCLCs from ccRCC and NK before and after R2TNF treatment. Responses to TNF were assessed by flow cytometry (FACS), immunofluorescence, and quantitative real-time PCR, TUNEL, and cell viability assays. Cytotoxic effect of CP was analyzed by Annexin V and propidium iodide staining with FACS. In addition, we assessed the effect of TNF on isolated SCLCs differentiation using a three-dimensional (3D) culture system. Clinical samples of ccRCC contain a greater number SCLCs compared to NK and the number of SCSC increases with higher tumor grade. Isolated SCLCs show expression of stemness markers (oct4, Nanog, Sox2, Lin28) but not differentiation markers (cytokeratin, CD31, CD45, and EpCAM). In ccRCC organ cultures, wtTNF and R2TNF increase CD133 and TNFR2 expression and promote cell cycle entry whereas wtTNF and R1TNF increase TNFR1 expression and promote cell death of SCLCs. Similar findings are observed in SCLCs isolated from NK but the effect was greater in SCLCs isolated from ccRCC. Application of CP distinctly triggered apoptotic and necrotic cell death in SLCSs pre-treatment with R2TNF as compared to CP treatment alone, with SCLCs from ccRCC more sensitive to CP compared to SLCS from NK. Furthermore, TNF promotes differentiation of SCLCs to an epithelial phenotype in 3D cultures, confirmed by cytokeratin expression and loss of stemness markers Nanog and Sox2. The differentiated cells show positive expression of TNF and TNFR2. These findings provide evidence that selective engagement of TNFR2 drive CSCs to cell proliferation/differentiation, and targeting of cycling cells with TNFR2 agonist in combination with anti-cancer agents may be a potential therapy for RCC.

Keywords: cancer stem cells, ccRCC, cell cycle, cell death, TNF, TNFR1, TNFR2, CD133

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Authors: N. Cetinkaya, F. Erdem

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Juncus acutus (Juncaceae) is a perennial wetland plant and it is commonly known as spiny rush or sharp rush. It is the most abundant plant in Kizilirmak grassland, Samsun, Turkey. Heavy metals are significant environmental contaminants in delta and their toxicity is an increasing problem for animals whose natural habitat is delta. The objective of this study was to evaluate heavy metal concentrations mainly As, Cd, Sb, Ba, Pb and Hg in stem and seed of Juncus acutus for grazing animals and birds in delta. The Juncus acutus stem and seed samples were collected from Kizilirmak Delta in July, August and September. Heavy metal concentrations of collected samples were analyzed by Inductively Coupled Plasma – Mass Spectrometer (ICP-MS). The obtained mean values of three months for As, Cd, Sb, Ba, Pb and Hg of stem and seed samples of Juncus acutus were 0.11 and 0.23 mg/kg; 0.07 and 0.11 mg/kg; 0.02 and 0.02 mg/kg; 5.26 and 1.75 mg/kg; 0.05 and not detectable in July respectively. Hg was not detected in both stem and seed of Juncus acutus, Pb concentration was determined only in stem of Juncus acutus but not in seed. There were no significant differences between the values of three months for As, Cd, Sb, Ba, Pb and Hg of stem and seed samples of Juncus acutus. The obtained As, Cd, Sb, Ba, Pb and Hg results of stem and seed of Juncus acutus show that seed and stem of Juncus acutus may be safely consumed for grazing animals and birds regarding to heavy metals contamination in Kizilirmak Delta.

Keywords: heavy metals, Juncus acutus, Kizilirmak Delta, wetland

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Authors: Natália Maria Rego

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ALeibnizn-algebraGis aK-vector space endowed whit a n-linearbracket operation [-,…-] : GG … G→ Gsatisfying the fundamental identity, which can be expressed saying that the right multiplication map Ry2, …, ᵧₙ: Gn→ G, Rᵧ₂, …, ᵧₙn(ˣ¹, …, ₓₙ) = [[ˣ¹, …, ₓₙ], ᵧ₂, …, ᵧₙ], is a derivation. This structure, together with its skew-symmetric version, named as Lie n-algebra or Filippov algebra, arose in the setting of Nambumechanics, an n-ary generalization of the Hamiltonian mechanics. Thefirst goal of this work is to provide a characterization of various classes of central extensions of Leibniz n-algebras in terms of homological properties. Namely, Commutator extension, Quasi-commutator extension, Stem extension, and Stem cover. These kind of central extensions are characterized by means of the character of the map *(E): nHL1(G) → M provided by the five-term exact sequence in homology with trivial coefficients of Leibniz n-algebras associated to an extension E : 0 → M → K → G → 0. For a free presentation 0 →R→ F →G→ 0of a Leibniz n-algebra G,the term M(G) = (R[F,…n.., F])/[R, F,..n-1..,F] is called the Schur multiplier of G, which is a Baer invariant, i.e., it does not depend on the chosen free presentation, and it is isomorphic to the first Leibniz n-algebras homology with trivial coefficients of G. A central extension of Leibniz n-algebras is a short exact sequenceE : 0 →M→K→G→ 0such that [M, K,.. ⁿ⁻¹.., K]=0. It is said to be a stem extension if M⊆[G, .. n.., G]. Additionally, if the induced map M(K) → M(G) is the zero map, then the stem extension Eis said to be a stem cover. The second aim of this work is to analyze the interplay between stem covers of Leibniz n-algebras and the Schur multiplier. Concretely, in the case of finite-dimensional Leibniz n-algebras, we show the existence of coverings, and we prove that all stem covers with finite-dimensional Schur multiplier are isoclinic. Additionally, we characterize stem covers of perfect Leibniz n-algebras.

Keywords: leibniz n-algebras, central extensions, Schur multiplier, stem cover

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Authors: Wing Mui Winnie So, Tian Luo, Zeyu Han

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To fully explore and develop the potentials of gifted students systematically and strategically by providing them with opportunities to receive education at appropriate levels, schools in Hong Kong are encouraged to adopt the "Three-Tier Implementation Model" to plan and implement the school-based gifted education, with Level Three refers to the provision of learning opportunities for the exceptionally gifted students in the form of specialist training outside the school setting by post-secondary institutions, non-government organisations, professional bodies and technology enterprises. Due to the growing concern worldwide about low interest among students in pursuing STEM (Science, Technology, Engineering, and Mathematics) careers, cultivating and boosting STEM career interest has been an emerging research focus worldwide. Although numerous studies have explored its critical contributors, little research has examined the effectiveness of comprehensive interventions such as “Studying with STEM professional”. This study aims to examine the effect on gifted students’ career interest during their participation in an off-school support programme designed and supervised by a team of STEM educators and STEM professionals from a university. Gifted students were provided opportunities and tasks to experience STEM career topics that are not included in the school syllabus, and to experience how to think and work like a STEM professional in their learning. Participants involved 40 primary school students joining the intervention programme outside the normal school setting. Research methods included adopting the STEM career interest survey and drawing tasks supplemented with writing before and after the programme, as well as interviews before the end of the programme. The semi-structured interviews focused on students’ views regarding STEM professionals; what’s it like to learn with a STEM professional; what’s it like to work and think like a STEM professional; and students’ STEM identity and career interest. The changes in gifted students’ STEM career interest and its well-recognised significant contributors, for example, STEM stereotypes, self-efficacy for STEM activities, and STEM outcome expectation, were collectively examined from the pre- and post-survey using T-test. Thematic analysis was conducted for the interview records to explore how studying with STEM professional intervention can help students understand STEM careers; build STEM identity; as well as how to think and work like a STEM professional. Results indicated a significant difference in STEM career interest before and after the intervention. The influencing mechanism was also identified from the measurement of the related contributors and the analysis of drawings and interviews. The potential of off-school support programme supervised by STEM educators and professionals to develop gifted students’ STEM career interest is argued to be further unleashed in future research and practice.

Keywords: gifted students, STEM career, STEM education, STEM professionals

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Authors: Mimoun Melliti

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This research paper investigates the promises, challenges, and perspectives associated with teaching STEM subjects in English within Tunisian higher education institutions. The study explores the potential benefits of English-medium instruction in Science, Technology, Engineering, and Mathematics (henceforth STEM) education, with a special focus on enhanced global competitiveness, improved English language proficiency, and increased access to international resources and collaborations. Additionally, it examines the various challenges faced by educators and students, such as language shift/barriers, curriculum adaptation, faculty training, and student support. Through a comprehensive analysis of promises and challenges, this paper aims to provide insights and recommendations for effectively implementing English medium instruction (henceforth EMI) of STEM in Tunisian higher education institutions. The paper concludes with a recommended action plan for the proper introduction of EMI of STEM in Tunisia.

Keywords: EMI;, STEM education, EFL, language reforms

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Authors: Yonghwa Lee, Yoon Suk Kim, Yongsub Yi

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This study was to confirm the effects of anti-melanogenesis and anti-inflammatory effects from Opuntia humifusa fruit and stem extracts. A potent anti-oxidant activity was shown from the leaf extract at IC50 value of 38.33±1.07 μg/mL and fruit extract at IC50 value of 40.23±2.21 μg/mL by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. Also, phenolic contents were confirmed total phenolic assay by high performance liquid chromatography (HPLC). Fraction of taxifolin from leaf extract was identified using HPLC and gas chromatography/mass spectrometry. The extracts of Opuntia humifusa fruit and stem were confirmed about toxicity effect in B16 F1 by cell viability. Melanin contents were decreased. Opuntia humifusa fruit and stem extracts had a positive effect of melanin synthesis inhibition for skin whitening. In investigating the anti-inflammatory activities of Opuntia humifusa, the results of cell viability indicated that taxifolin did not show cytotoxicity on RAW264.7 cells at 500 μM of concentration. The results show that taxifolin inhibited lipopolysaccharide (LPS)-induced production of Nitrite oxide (NO). In addition, taxifolin indicated the inhibition of lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF) -α and interleukin (IL) -6 productions by cytokine assay and cyclooxygenase (COX)-2 expression by western blot analysis, meaning that taxifolin had a significant anti-inflammatory effect. Our results suggested that taxifolin from Opuntia humifusa has anti-melanogenesis and anti-inflammatory activities.

Keywords: anti-melanogenesis, anti-inflammatory, Opuntia humifusa, taxifolin

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Authors: Tian Luo, Wing Mui Winnie So

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Gaining career awareness in STEM is an important educational outcome in STEM education. While many studies focused on students’ understanding of scientists and engineers, very few studies explore students’ perceptions of technologists as a group of STEM professionals. In this study, 300 valid surveys which include drawing task and follow-up questions about technologist were collected from 4th to 6th grade students. The results showed that 75.1% of the students draw a technologist as a male and 19.3% draw a technologist as a female. Most students believe that technologists use math, science or engineering in their work and can name a few categories of technologists. The drawings also showed that students tend to present technologists as people who work with a computer.

Keywords: STEM (Science, Technology, Engineering, and Mathematics) education, elementary students, technologist, STEM careers

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Authors: Roman Major, Klaudia Trembecka- Wojciga, Juergen Markus Lackner, Boguslaw Major

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The future and the development of science is therefore seen in interdisciplinary areas such as bio medical engineering. Self-assembled structures, similar to stem cell niches would inhibit fast division process and subsequently capture the stem cells from the blood flow. By means of surface topography and the stiffness as well as micro structure progenitor cells should be differentiated towards the formation of endothelial cells monolayer which effectively will inhibit activation of the coagulation cascade. The idea of the material surface development met the interest of the clinical institutions, which support the development of science in this area and are waiting for scientific solutions that could contribute to the development of heart assist systems. This would improve the efficiency of the treatment of patients with myocardial failure, supported with artificial heart assist systems. Innovative materials would enable the redesign, in the post project activity, construction of ventricular heart assist.

Keywords: bio-inspired materials, electron microscopy, haemocompatibility, niche-like structures, thin coatings

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Authors: Daniel Aberdam

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Haploinsufficiency of PAX6 in humans is the main cause of congenital aniridia, a rare eye disease characterized by reduced visual acuity. Patients have also progressive disorders including cataract, glaucoma and corneal abnormalities making their condition very challenging to manage. Aniridia-related keratopathy (ARK), caused by a combination of factors including limbal stem-cell deficiency, impaired healing response, abnormal differentiation, and infiltration of conjunctival cells onto the corneal surface, affects up to 95% of patients. It usually begins in the first decade of life resulting in recurrent corneal erosions, sub-epithelial fibrosis with corneal decompensation and opacification. Unfortunately, current treatment options for aniridia patients are currently limited. Although animal models partially recapitulate this disease, there is no in vitro cellular model of AKT needed for drug/therapeutic tools screening and validation. We used genome editing (CRISPR/Cas9 technology) to introduce a nonsense mutation found in patients into one allele of the PAX6 gene into limbal stem cells. Resulting mutated clones, expressing half of the amount of PAX6 protein and thus representative of haploinsufficiency were further characterized. Sequencing analysis showed that no off-target mutations were induced. The mutated cells displayed reduced cell proliferation and cell migration but enhanced cell adhesion. Known PAX6 targets expression was also reduced. Remarkably, addition of soluble recombinant PAX6 protein into the culture medium was sufficient to activate endogenous PAX6 gene and, as a consequence, rescue the phenotype. It strongly suggests that our in vitro model recapitulates well the epithelial defect and becomes a powerful tool to identify drugs that could rescue the corneal defect in patients. Furthermore, we demonstrate that the homeotic transcription factor Pax6 is able to be uptake naturally by recipient cells to function into the nucleus.

Keywords: Pax6, crispr/cas9, limbal stem cells, aniridia, gene therapy

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Authors: Kumari Rekha, Mathur K Dhananjay, Maheshwari Deepanshu, Nautiyal Nidhi, Shubham Smriti, Laal Deepika, Sinha Swati, Kumar Anupam, Biswas Subhrajit, Shiv Kumar Sarin

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Background: Mesenchymal stem cells are multipotent stem cells derived from mesoderm and are used for therapeutic purposes because of their self-renewal, homing capacity, Immunomodulatory capability, low immunogenicity and mitochondrial transfer signaling. MSCs have the ability to regulate the mechanism of both innate as well as adaptive immune responses through the modulation of cellular response and the secretion of inflammatory mediators. Different sources of MSC are UC MSC, BM MSC, Dental Pulp, and Adipose MSC. The most frequent source used is umbilical cord tissue due to its being easily available and free of limitations of collection procedures from respective hospitals. The immunosuppressive role of MSCs is particularly interesting for clinical use since it confers resistance to rejection by the host immune response. Methodology: In this study, T helper cells (TH4), Cytotoxic T cells (CD-8), immunoregulatory cells (CD25 +FOXP3+) are compared from isolated MSC from three different methods, UC Dissociation Kit (Miltenyi), Explant Culture and Collagenase Type-IV. To check the immunomodulatory property, these MSCs were seeded with PBMC(Coculture) in CD3 coated 24 well plates. Cd28 antibody was added in coculture for six days. The coculture was analyzed in FACS Verse flow cytometry. Results: From flow cytometry analysis of coculture, it found that All over T helper cells (CD4+) number p<0.0264 increases in (All Enzymes) MSC rather than explant MSC(p>0.0895) as compared to Collagenase(p>0.7889) in a coculture of Activated T cell and Mesenchymal Stem Cell. Similar T reg cells (CD25+, FOXP3+) expression p<0.0234increases in All Enzymes), decreases in Explant and Collagenase. Experiments have shown that MSCs can also directly prevent the cytotoxic activity of CD8 lymphocytes mainly by blocking their proliferation rather than by inhibiting the cytotoxic effect. And promoting the t-reg cells, which helps in the mediation of immune response in various diseases. Conclusion: MSC suppress Cytotoxic CD8 T cell and Enhance immunoregulatory T reg (CD4+, CD25+, FOXP3+) Cell expression. Thus, MSC maintains a proper balance(ratio) between CD4 T cells and Cytotoxic CD8 T cells.

Keywords: MSC, disease, T cell, T regulatory

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Authors: Elena Filova, Ondrej Kaplan, Marie Markova, Helena Dragounova, Roman Matejka, Eduard Brynda, Lucie Bacakova

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Decellularized vessels have been evaluated as small-diameter vascular prostheses. Reseeding autologous cells onto decellularized tissue prior implantation should prolong prostheses function and make them living tissues. Suitable cell types for reseeding are both endothelial cells and bone marrow-derived stem cells, with a capacity for differentiation into smooth muscle cells upon mechanical loading. Endothelial cells assure antithrombogenicity of the vessels and MSCs produce growth factors and, after their differentiation into smooth muscle cells, they are contractile and produce extracellular matrix proteins as well. Fibrin is a natural scaffold, which allows direct cell adhesion based on integrin receptors. It can be prepared autologous. Fibrin can be modified with bound growth factors, such as basic fibroblast growth factor (FGF-2) and vascular endothelial growth factor (VEGF). These modifications in turn make the scaffold more attractive for cells ingrowth into the biological scaffold. The aim of the study was to prepare thin surface-attached fibrin assemblies with bound FGF-2 and VEGF, and to evaluate growth and differentiation of bone marrow-derived mesenchymal stem cells on the fibrin (Fb) assemblies. Following thin surface-attached fibrin assemblies were prepared: Fb, Fb+VEGF, Fb+FGF2, Fb+heparin, Fb+heparin+VEGF, Fb+heparin+FGF2, Fb+heparin+FGF2+VEGF. Cell culture poly-styrene and glass coverslips were used as controls. Human MSCs (passage 3) were seeded at the density of 8800 cells/1.5 mL alpha-MEM medium with 2.5% FS and 200 U/mL aprotinin per well of a 24-well cell culture. The cells have been cultured on the samples for 6 days. Cell densities on day 1, 3, and 6 were analyzed after staining with LIVE/DEAD cytotoxicity/viability assay kit. The differentiation of MSCs is being analyzed using qPCR. On day 1, the highest density of MSCs was observed on Fb+VEGF and Fb+FGF2. On days 3 and 6, there were similar densities on all samples. On day 1, cell morphology was polygonal and spread on all sample. On day 3 and 6, MSCs growing on Fb assemblies with FGF2 became apparently elongated. The evaluation of expression of genes for von Willebrand factor and CD31 (endothelial cells), for alpha-actin (smooth muscle cells), and for alkaline phosphatase (osteoblasts) is in progress. We prepared fibrin assemblies with bound VEGF and FGF-2 that supported attachment and growth of mesenchymal stem cells. The layers are promising for improving the ingrowth of MSCs into the biological scaffold. Supported by the Technology Agency of the Czech Republic TA04011345, and Ministry of Health NT11270-4/2010, and BIOCEV – Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University” project (CZ.1.05/1.1.00/02.0109), funded by the European Regional Development Fund for their financial supports.

Keywords: fibrin assemblies, FGF-2, mesenchymal stem cells, VEGF

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Authors: Raluca Ionela Maxim

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Designing a gender responsive Science, Technology, Engineering and Mathematics (STEM) mobile game based learning solution (mGBL) is a challenge in terms of content, gamification level and equal engagement of girls and boys. The goal of this case study was to research and create a high-fidelity prototype design of a mobile game that contains role-models as avatars that guide and expose girls and boys to STEM learning content. For this research purpose it was applied the methodology of design sprint with five-phase process that combines design thinking principles. The technique of this methodology comprises smart interviews with STEM experts, mind-map creation, sketching, prototyping and usability testing of the interactive prototype of the gender responsive STEM mGBL. The results have shown that the effect of the avatar/role model had a positive impact. Therefore, by exposing students (boys and girls) to STEM role models in an mGBL tool is helpful for the decreasing of the gender inequalities in STEM fields.

Keywords: design thinking, design sprint, gender-responsive STEM education, mobile game based learning, role-models

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Authors: Qurratulain Faheem

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The human body often serves as a reference point to analyse the notions of self and society. Situating on Merleau-Ponty and Bourdieu theories of embodiments, this research explores the notions around the human body and its influence on the ethical considerations in regards to organ transplantation among the Muslim communities in Pakistan. The context of Pakistan makes an intriguing case study as cadaveric organ transplantation is not in practise. Whereas living organ transplantation is commonly is practised between family membersonly. These contradictory practices apparently rests on the ideologies around the human body and religious beliefs as well the personal judgements and authority of healthcare professionals. This research is a year-long ethnographic study carried out as part of doctoral studies. An anthropological approach towards organ transplantation in Pakistan brought forward various socio-cultural notions around the human body and selfhood that serve as a framework around biomedical ethical issues in various societies. Further, it surface the contradictions and issues associated with organ transplantation that makes it a dilemma situated in a nexus of various socio-cultural and political factors rather seeing it as an isolated health concern. This research is a novel study on the subject of organ transplantation in the context of Pakistan but also put forward ethnographic data that could serve as a reference in other religious societies. Further, the ethnographic data bring forward experiences and stories of organ receivers, organ donors, religious leaders, healthcare professionals, and the general public, which aspire to encourage biomedical ethicists and social-scientists to consider ethnography as a research methodology and rely upon people’s lived experiences while establishing policies and practices around biomedical ethical issues.

Keywords: organ transplantation, ethics, pakistan, gender, islam, muslims, living organ donation

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Authors: Annette McArthur

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The traditional model of teaching and learning, where ICT sits as a separate entity is not a model for a 21st century school. It is imperative that teaching and learning embraces technological advancements. The challenge in schools lies in shifting the mindset of teachers so they see ICT as integral to their teaching, learning and curriculum rather than a separate E-Learning curriculum stream. This research project investigates how the effective, planned, intentional integration of ICT into a STEM curriculum, can enable the shift in the teacher mindset. The project incorporated: • Developing a professional coaching relationship with key STEM teachers. • Facilitating staff professional development involving student centered project based learning pedagogy in the context of a STEM curriculum. • Facilitating staff professional development involving digital literacy. • Establishing a professional community where collaboration; sharing and reflection were part of the culture of the STEM community. • Facilitating classroom support for the effective delivery innovative STEM curriculum. • Developing STEM learning spaces where technologies were used to empower and engage learners to participate in student-centered, project-based learning.

Keywords: e-learning, ICT, project based learning, STEM

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Authors: Joseph P. Mandala

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This paper examines the legal and medical ethics concerns, which proponents of human head transplantation continue to defy since the procedure was first attempted on dogs in 1908. Despite recent bioethical objections, proponents have proceeded with radical experimentation, claiming transplantation would treat incurable diseases and improve patients’ quality of life. In 2018, Italian neurosurgeon, Sergio Canavero, and Dr. Xiaoping Ren claimed to have performed a head transplant on a corpse in China. Content analysis of literature shows that the procedure failed to satisfy scientific, legal, and bioethical elements because, unlike humans, corpses cannot coordinate function. Putting a severed head onto a body that has been dead for several days is not equivalent to a transplant which would require successfully reconnecting and restoring function to a spinal cord. While reconnection without restoration of bodily function is not transplantation, the publicized procedure on animals and corpses could leapfrog to humans, sparking excitement in society likely to affect organ donors and recipients from territorial jurisdictions with varying legal and ethical regimes. As neurodiscoveries generate further excitement, the need to preemptively address the legal and medical ethics impact of head transplantation in our society cannot be overstated. A preemptive development of methods to address the impact of head transplantation will help harmonizing national and international laws on organ donations, advance directives, and laws affecting end of life.

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Authors: Josephine O. Okocha, Ifeanyi Adigwe

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This research aims to examine the factors impacting women's retention in STEM in Nigeria and Georgia. In a bid to come up with strategies to enhance women’s participation in STEM, this study identifies and juxtaposes the factors impacting the retention of women in STEM and how they vary from one country to another are discussed. This study adopted the literature review method to perform the critical analysis. A total of 76 papers were retrieved from the Scopus database and were published between 2018 and 2023. Only 12 papers met the criteria for inclusion in the analysis. The findings reveal that the factors impacting women’s retention in STEM include funding (NGOs and government agencies), scholarship, specialized recruitment, mentoring, the establishment of women-only higher institutions, creating a balanced work and family environment, combating stereotypes, and enabling policies and laws. The paper highlights some key recommendations to help improve the retention of women in STEM in Africa and Nigeria in particular.

Keywords: STEM, women, retention, career, Nigeria, Georgia, women’s retention, women representation

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Authors: Niklas Ravn-Boess, Nainita Bhowmick, Takamitsu Hattori, Shohei Koide, Christopher Park, Dimitris Placantonakis

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Background: Glioblastoma (GBM) is the most common and deadly primary brain malignancy in adults. Tumor propagation, brain invasion, and resistance to therapy critically depend on GBM stem-like cells (GSCs); however, the mechanisms that regulate GSC self-renewal are incompletely understood. Given the aggressiveness and poor prognosis of GBM, it is imperative to find biomarkers that could also translate into novel drug targets. Along these lines, we have identified a cell surface antigen, CD97 (ADGRE5), an adhesion G protein-coupled receptor (GPCR), that is expressed on GBM cells but is absent from non-neoplastic brain tissue. CD97 has been shown to promote invasiveness, angiogenesis, and migration in several human cancers, but its frequency of expression and functional role in regulating GBM growth and survival, and its potential as a therapeutic target has not been investigated. Design: We assessed CD97 mRNA and protein expression in patient derived GBM samples and cell lines using publicly available RNA-sequencing datasets and flow cytometry, respectively. To assess CD97 function, we generated shRNA lentiviral constructs that target a sequence in the CD97 extracellular domain (ECD). A scrambled shRNA (scr) with no predicted targets in the genome was used as a control. We evaluated CD97 shRNA lentivirally transduced GBM cells for Ki67, Annexin V, and DAPI. We also tested CD97 KD cells for their ability to self-renew using clonogenic tumorsphere formation assays. Further, we utilized synthetic Abs (sAbs) generated against the ECD of CD97 to test for potential antitumor effects using patient-derived GBM cell lines. Results: CD97 mRNA expression was expressed at high levels in all GBM samples available in the TCGA cohort. We found high levels of surface CD97 protein expression in 6/6 patient-derived GBM cell cultures, but not human neural stem cells. Flow cytometry confirmed downregulation of CD97 in CD97 shRNA lentivirally transduced cells. CD97 KD induced a significant reduction in cell growth in 3 independent GBM cell lines representing mesenchymal and proneural subtypes, which was accompanied by reduced (~20%) Ki67 staining and increased (~30%) apoptosis. Incubation of GBM cells with sAbs (20 ug/ ml) against the ECD of CD97 for 3 days induced GSC differentiation, as determined by the expression of GFAP and Tubulin. Using three unique GBM patient derived cultures, we found that CD97 KD attenuated the ability of GBM cells to initiate sphere formation by over 300 fold, consistent with an impairment in GSC self-renewal. Conclusion: Loss of CD97 expression in patient-derived GBM cells markedly decreases proliferation, induces cell death, and reduces tumorsphere formation. sAbs against the ECD of CD97 reduce tumorsphere formation, recapitulating the phenotype of CD97 KD, suggesting that sAbs that inhibit CD97 function exhibit anti-tumor activity. Collectively, these findings indicate that CD97 is necessary for the proliferation and survival of human GBM cells and identify CD97 as a promising therapeutically targetable vulnerability in GBM.

Keywords: adhesion GPCR, CD97, GBM stem cell, glioblastoma

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Authors: Hsin-Lien Lin, Ju-Hui Fu

Abstract:

Dendritic cells (DCs) are the major professional antigen-presenting cells for the development of optimal T-cell immunity. DCs can be used as pharmacological targets to screen novel biological modifiers for the treatment of harmful immune responses, such as transplantation rejection. Dryopteris crassirhizoma Nakai (Aspiadaceae) is used for traditional herbal medicine in the region of East Asia. The root of this fern plant has been listed for treating inflammatory diseases. Dryocrassin is the tetrameric phlorophenone component derived from Dryopteris. Here, we tested the immunomodulatory potential of dryocrassin on lipopolysaccharide (LPS)-stimulated activation of mouse bone marrow-derived DCs in vitro and in skin allograft transplantation in vivo. Results demonstrated that dryocrassin reduced the secretion of tumor necrosis factor-α, interleukin-6, and interleukin-12p70 by LPS-stimulated DCs. The expression of LPS-induced major histocompatibility complex class II, CD40, and CD86 on DCs was also blocked by dryocrassin. Moreover, LPS-stimulated DC-elicited allogeneic T-cell proliferation was lessened by dryocrassin. In addition, dryocrassin inhibited LPS-induced activation of IϰB kinase, JNK/p38 mitogen-activated protein kinase, as well as the translocation of NF-ϰB. Treatment with dryocrassin obviously diminished 2,4-dinitro-1-fluorobenzene- induced delayed-type hypersensitivity and prolonged skin allograft survival. Dryocrassin may be one of the potent immunosuppressive agents for transplant rejection through the destruction of DC maturation and function.

Keywords: dryocrassin, dendritic cells, immunosuppression, skin allograft

Procedia PDF Downloads 353

Authors: Reyhane Hamed Kamran

Abstract:

Early stage morphogenesis requires the execution of complex systems that direct the nearby conduct of gatherings of cells. The organization of such instruments has been, for the most part, deciphered through the recognizable proof of moderated groups of flagging pathways that spatially and transiently control cell conduct. In any case, how this data is handled to control cell shape and cell elements is an open territory of examination. The structure that rises up out of differing controls, for example, cell science, material science, and formative science, focuses to bond and cortical actin arranges as controllers of cell surface mechanics. In this specific circumstance, a scope of formative marvels can be clarified by the guideline of cell surface pressure.

Keywords: cell, tissue damage, morphogenesis, cell conduct

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Authors: Narin Salehiyan

Abstract:

Early stage morphogenesis requires the execution of complex systems that direct the nearby conduct of gatherings of cells. The organization of such instruments has been, for the most part, deciphered through the recognizable proof of moderated groups of flagging pathways that spatially and transiently control cell conduct. In any case, how this data is handled to control cell shape and cell elements is an open territory of examination. The structure that rises up out of differing controls, for example, cell science, material science and formative science, focuses to bond and cortical actin arranges as controllers of cell surface mechanics. In this specific circumstance, a scope of formative marvels can be clarified by the guideline of cell surface pressure.

Keywords: cell, tissue damage, morphogenesis, cell conduct

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Authors: Raza Ullah, Hazir Ullah

Abstract:

There is ample evidence that reveals the outstanding performance of girls in a different range of subjects. However, it is pertinent to mention here that boys have historically dominated girls, particularly in math, physics, and technological subjects across the globe with the exception of few developed countries. This article examines the reasons why girls are underdog in STEM subjects. The article critically analyzes two main approaches towards gender and education: biological determinist and feminist. This article highlights that social factors influencing girls performance in STEM subjects have not analyzed critically, and girls underachieving in science has linked with biological and sex differences. The article concludes that the underperformance of girls in a STEM subject is the direct response of socio-cultural factors. Thus, socio-cultural factors are responsible for the dearth of girls in STEM subjects.

Keywords: gender, underperformance, STEM, education, sex

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Authors: Shin-Yi Mao, Jiashing Yu

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Decellularized adipose tissue (DAT) has received lots of attention as biological scaffolds recently. DAT that extracted from the extracellular matrix (ECM) of adipose tissues holds great promise as a xenogeneic biomaterial for tissue engineering and regenerative medicine. In our study, 2-D DATsol film was fabricated to enhance cell adhesion, proliferation, and differentiation of ASCs in vitro. DAT was also used to modify alginate for improvement of cell adhesion. Alginate microspheres modified with DAT were prepared by Nisco. These microspheres could provide a highly supportive 3-D environment for ASCs. In our works, ASCs were immobilized in alginate microspheres modified with DAT to promoted cell adhesion and adipogenic differentiation. Accordingly, we hypothesize that tissue regeneration in vivo could be promoted with the aid of modified microspheres in future.

Keywords: adipose stem cells, decellularize adipose tissue, Alginate, microcarries

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Authors: R. Zhankina, U. Zhanbyrbekuly, A. Tamadon, M. Askarov, R. Sherkhanov, D. Akhmetov, D. Saipiyeva, N. Keulimzhaev

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Male infertility affects about 15% of couples of reproductive age. Approximately 10–15% have azoospermia who have previously been diagnosed with male infertility. Azoospermia is regarded as the absence of spermatozoa in the ejaculate and is found in 10-15% of infertile men. Non-obstructive azoospermia is considered a cause of male infertility that is not amenable to drug therapy. Patients with non-obstructive azoospermia are unable to have their "own" children and have only options for adoption or use of donor sperm. Advances in assisted reproductive technologies such as intracytoplasmic sperm injection in vitro fertilization have significantly changed the management of patients with non-obstructive azoospermia. Advances in biotechnology have increased the options for treating patients with non-obstructive azoospermia. Mesenchymal stem cell therapy has been recognized as a new option for infertility treatment. Material and methods of the study: After obtaining informed consent, 5 patients diagnosed with non-obstructive azoospermia were included in an open, non-randomized study. The age of the patients ranged from 24 to 35 years. The examination was carried out before the start of treatment, which included biochemical blood tests, hormonal profile levels (luteinizing hormone, follicle-stimulating hormone, testosterone, prolactin, inhibin B); tests for tumor markers; genetic research. All studies were carried out in compliance with the requirements of Protocol No. 8 dated 06/09/20, approved by the Local Ethical Commission of NJSC "Astana Medical University". The control examination of patients was carried out after 6 months, by re-taking the program and hormonal profile (testosterone, luteinizing hormone, follicle-stimulating hormone, prolactin, inhibin B). Before micro-TESE of the testis, all 5 patients underwent myeloexfusion in the operating room. During the micro-TESE, autotransplantation of mesenchymal stem cells into the testicular network, previously cultured in a cell technology laboratory for 2 weeks, was performed. Results of the study: in all patients, the levels of total testosterone increased, the level of follicle-stimulating hormone decreased, the levels of luteinizing hormone returned to normal, the level of inhibin B increased. IVF with a positive result; another patient (20%) had spermatogenesis cells. Non-obstructive azoospermia and mesenchymal stem cells Conclusions: The positive results of this work serve as the basis for the application of a new cellular therapeutic approach for the treatment of non-obstructive azoospermia using mesenchymal stem cells.

Keywords: cell therapy, regenerative medicine, male infertility, mesenchymal stem cells

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Authors: Hami Ashraf, Farid Kosari

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Graft-versus-host disease (GvHD) is a common complication of allogeneic hematopoietic stem cell transplantation that can lead to significant morbidity and mortality. Histopathological examination of affected tissues is an essential tool for diagnosing and grading GvHD in animal models, which are used to study disease mechanisms and evaluate new therapies. In this systematic review, we identified and analyzed original research articles in PubMed, Scopus, Web of Science, and Google Scholar that described grading systems for GvHD in animal models based on histopathological features. We found that several grading systems have been developed, which vary in the tissues and criteria they assess, the severity scoring scales they use, and the level of detail they provide. Skin, liver, and gut are the most commonly evaluated tissues, but lung and thymus are also included in some systems. Our analysis highlights the need for standardized criteria and consistent use of grading systems to enable comparisons between studies and facilitate the translation of preclinical findings to clinical practice.

Keywords: graft-versus-host disease, GvHD, animal model, histopathology, grading system

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Authors: Qurratulain Faheem

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The human body often serves as a reference point to analyse the notions of self and society. Situating on Merleau-Ponty and Bourdieu theories of embodiments, this research explores the notions around the human body and its influence on the ethical considerations in regards to organ transplantation among the Muslim communities in Pakistan. The context of Pakistan makes an intriguing case study as cadaveric organ transplantation is not in practise. Whereas living organ transplantation is commonly is practised between family members only. These contradictory practices apparently rests on the ideologies around the human body and religious beliefs as well the personal judgements and authority of healthcare professionals. This research is a year-long ethnographic study carried out as part of doctoral studies. An anthropological approach towards organ transplantation in Pakistan brought forward various socio-cultural notions around the human body and selfhood that serve as a framework around biomedical ethical issues in various societies. Further, it surface the contradictions and issues associated with organ transplantation that makes it a dilemma situated in a nexus of various socio-cultural and political factors rather seeing it as an isolated health concern. This research is a novel study on the subject of organ transplantation in the context of Pakistan but also put forward ethnographic data that could serve as a reference in other religious societies. Further, the ethnographic data bring forward experiences and stories of organ receivers, organ donors, religious leaders, healthcare professionals, and the general public, which aspire to encourage biomedical ethicists and social-scientists to consider ethnography as a research methodology and rely upon people’s lived experiences while establishing policies and practices around biomedical ethical issues.

Keywords: Gender, organ transplantation, muslims, pakistan, organ donation, bioethics, culture and religion, gender

Procedia PDF Downloads 77

Authors: Kholik

Abstract:

Nowadays, the demand of renewable energy in general is increasing due to the crisis of fossil fuels. Biohydrogen is an alternative fuel with zero emission derived from renewable resources such as banana pseudo stem of agricultural residues. Banana plant can be easily found in tropical and subtropical areas, so the resource is abundant and readily available as a biohydrogen substrate. Banana pseudo stem has not been utilised as a resource or substrate of biohydrogen production and it mainly contains 45-65% cellulose (α-cellulose), 5-15% hemicellulose and 20-30% Lignin, which indicates that banana pseudo stem will be renewable, sustainable and promising resource as lignocellulosic biomass. In this research, biohydrogen is derived from banana pseudo stem by dark fermentation. Dark fermentation is the most suitable approach for practical biohydrogen production from organic solid wastes. The process has several advantages including a fast reaction rate, no need of light, and a smaller footprint. 321 million metric tonnes banana pseudo stem of 428 million metric tonnes banana plantation residues in worldwide for 2013 and 22.5 million metric tonnes banana pseudo stem of 30 million metric tonnes banana plantation residues in Indonesia for 2015 will be able to generate 810.60 million tonne mol H2 and 56.819 million tonne mol H2, respectively. In this paper, we will show that the banana pseudo stem is the renewable, sustainable and promising resource to be utilised and to produce biohydrogen as energy generation with high yield and high contain of cellulose in comparison with the other substrates.

Keywords: banana pseudo stem, biohydrogen, dark fermentation, lignocellulosic

Procedia PDF Downloads 324