Biotechnology Commons^™

Metabolite Analysis To Understand Factors Limiting Microbial Fermentation At High Substrate Concentrations, Bishal D. Sharma

Dartmouth College Ph.D Dissertations

Clostridium thermocellum, a thermophilic anaerobic bacterium, holds promise as a biocatalyst for conversion of lignocellulosic biomass into biofuels. This microbe has been subjected to various metabolic engineering strategies, including the deletion of genes encoding secondary fermentation products such as H₂, formate, acetate, and lactate, as well as the expression of heterologous genes to redirect carbon flux toward ethanol production. Despite these efforts, achieving economically viable ethanol titers remains a challenge. We hypothesized that studying how intracellular metabolite concentrations change as fermentation stops could reveal metabolic bottlenecks or regulatory mechanisms limiting ethanol production at high titers. However, metabolomics studies at …

Effect Of Meta-Topolin And Kinetin At Various Concentrations On Shoot Multiplication In White Turmeric (Curcuma Zedoaria Rosc.), Murgayanti, Annisa Nanda Nur Azizah, Anne Nuraini, Erni Suminar, Syariful Mubarok

Jurnal Kultivasi

White turmeric is a medicinal plant widely used to treat various diseases. The limited availability of white turmeric seedlings is influenced by the scarcity of quality seeds due to the long dormancy of the rhizomes and the high incidence of pathogen attacks. Tissue culture techniques are one alternative to address the problem of white turmeric seedling availability. This study aims to determine the best concentrations of meta-topolin and kinetin for in vitro shoot multiplication of white turmeric. The research was conducted from January to June 2022 at the tissue culture laboratory, Faculty of Agriculture, Universitas Padjadjaran. This study used a …

Crispr-Induced Mutagenesis Of Arabidopsis Thaliana Gene Apetala3, Hazel Frans, Tara Phelps-Durr

SACAD: Scholarly Activities

This project aims to create CRISPR-CAS9 mutations in the APETELA3 (AP3) gene of the model plant Arabidopsis thaliana. AP3 is a class B gene critical to the petal and stamen development of Arabidopsis flowers. AP3 is defined in a MADS domain, which binds directly to DNA and may be responsible for the expression of the CaRG-box genes. AP3 works in conjunction with PISTILLATA (PI), AGAMOUS (AG), APETALA1 (AP1), and SEPALLATA (SEP) genes to specify the development in the second and third whorls of the flower. While several alleles of AP3 already exist, these alleles are strong alleles that knockout gene …

The Design, Construction, And Testing Of Mrna Vaccine Against Equine Herpes Virus, Bhawana Devkota

LSU Master's Theses

Equine Herpesvirus-1 (EHV 1) is a worldwide significant pathogen that causes respiratory illness, abortion, and neurological disorders in equine. Current vaccines, including live attenuated, inactivated, and subunit platforms, do not prevent viral latency, mucosal shedding, or cross-strain immunity, requiring alternative approaches. To address these gaps, this project explores the design, synthesis, and in vitro testing of an mRNA vaccine targeting immunogenic EHV 1 glycoprotein (gB, gC, gD, gG, and gM). Epitopes were computationally predicted using the Immune Epitope Database (IEDB), codon-optimized for Bos taurus, and cloned into pUCIDT vectors using SP6/T7 promoters. In vitro transcription (IVT) used nucleotide modifications …

Development Of A Multi-Epitope Peptide Vaccine Against Monkeypox Virus: Immunoinformatics Analysis For South East Asian Hla Alleles, Nelson Chandra, Mochammad Aqilah Herdiansyah, Viol Dhea Kharisma, Arif Nur Muhammad Ansori, Arli Aditya Parikesit

Makara Journal of Science

The monkeypox virus (MPXV), a DNA virus causing zoonotic disease, poses major global public health challenges, with mortality rates between 3%–6%. Although smallpox vaccines provide partial cross-protection, there is a critical need for a dedicated, effective monkeypox (mpox) vaccine. This study aimed to design a multi-epitope peptide-based vaccine specifically adapted to the HLA allele profiles common in Southeast Asian populations, where MPXV cases are rising. Using immunoinformatics, we screened for and detected B and T cell epitopes from the MPXV cell surface antigen and IFN-alpha/beta receptor proteins. The vaccine design was validated through a rigorous evaluation of its antigenicity, immunogenicity, …

Cloning And Expression Of Scamp3 In Escherichia Coli Bl21(De3) With In Silico Sequence-Based Cancer Epitopes Prediction, Selly Setiati Rajagukguk, Sabar Pambudi, Astari Dwiranti, Doddy Irawan Setyo Utomo, Anom Bowolaksono

Makara Journal of Science

Secretory carrier membrane protein 3 (SCAMP3) is a crucial membrane protein involved in intracellular vesicle traffick-ing and exocytosis. The SCAMP3 expression has been observed in diverse cancer types, such as melanoma, glioma, hepatocellular and breast cancer. Increased SCAMP3 expression has been reported in certain cancer cells relative to that in normal cells, suggesting the potential role of SCAMP3 in cancer development or progression. In this study, we successfully cloned and expressed SCAMP3 in Escherichia coli strain BL21(DE3). SCAMP3 was amplified and insert-ed directionally into the prokaryotic expression vector pET21d(+). The transformation of recombinant plasmid into E. coli BL21(DE3) cells were …

Screening For Penicillin G Acylase (Pga)-Producing Bacteria And Gene Cloning Using Degenerate Oligonucleotide Primed-Pcr, Masdalifah Masdalifah, Sri Rezeki Wulandari, Gabriela Christy Sabbathini, Maria Ulfah, Dini Achnafani, Ahmad Wibisana, Feronika Heppy Sriherfyna, Is Helianti, Niknik Nurhayati

Makara Journal of Science

The growing concern over antibiotic resistance has driven global efforts to explore innovative solutions, including the use of Penicillin G acylase (PGA) to produce semisynthetic β-lactam antibiotics. This study screened four potential in-tracellular PGA-producing bacteria: Alcaligenes faecalis InaCC B444 (AfPGA), Kluyvera cryocrescens InaCC B850 (KcPGA), Providencia rettgeri InaCC B25 (Pr25PGA), and P. rettgeri InaCC B466 (Pr466PGA). Penicillin G Acylase encoding genes (pgas) were isolated from them using a Degenerate Oligonucleotide Primed-PCR (DOP-PCR) approach and sequenced. Microbiological assays confirmed all tested crude extracts to exhibit inhibitory effects. Penicillin G was used for evaluating hydrolytic activity and 6-Amino Penicillanic Acid (6-APA) coupled …

Curcumin Attenuates Smoking And Drinking Activated Nf-Κb/Il-6 Inflammatory Signaling Axis In Cervical Cancer, Vivek K. Kashyap, Prashanth Kb Nagesh, Ajay K. Singh, Andrew Massey, Godwin P. Darkwah, Aaron George, Sheema Khan, Nadeem Zafar, Santosh Kumar, Namita Sinha, Meena Jaggi, Subhash C. Chauhan

Research Symposium

Background: High-risk strains of HPV are known to cause cervical cancer. Multiple clinical studies have emphasized that smoking and drinking are critical risk factors for cervical cancer and its high-grade precursors. In this study, we investigated if smoking and/or drinking augment the molecular mechanisms of the cervical carcinogenesis and defined a potential therapeutic approach for their attenuation.

Methods: The impact of benzo[a]pyrene (B[a]P) and/or ethanol (EtOH) exposure on cervical cancer cells was assessed by measuring changes in their cell migration and invasion characteristics. Expression of HPV16 E6/E7, NF-κB, cytokines, and inflammation mediators was determined using qRT-PCR, immunoblotting, ELISA, luciferase reporter …

Comparative Evaluation Of Bacterial Adhesion And Biofilm Formation On Contact Lenses, Lina M. Shaker, Wan Nor Roslam Wan Isahak

AUIQ Complementary Biological System

This study evaluates bacterial adhesion, biofilm formation, and Staphylococcus aureus (S. aureus) (ATCC 25923) colonization on medical and cosmetic contact lenses made from Hilafilcon B and Omafilcon B under simulated ocular conditions. Non-incubated lenses served as controls, and experiments were repeated three times with a variability margin of ±5%, ensuring accuracy and reliability. Results revealed that biofilm growth and bacterial adhesion were influenced by lens material, type, and incubation duration. Cosmetic lenses made from Hilafilcon B exhibited the highest biofilm growth (1.95%) after one day, highlighting their susceptibility to microbial colonization. Conversely, medical lenses made from Hilafilcon B …

Removal Of Cr(Vi) From Aqueous Solution Using Magnetic Activated Carbon Prepared From Waste Shell Of Marigold Flower Straw: Kinetics, Isotherm And Mechanism Studies, Shanmukha Rao Metta, Uttam Kumar Sahu

AUIQ Complementary Biological System

When Cr(VI) was released from industrial waste into water bodies harms aquatic ecosystems and poses risks to human well-being. To reduce risks, it's crucial to lower Cr(VI) levels through the sustainable process. In order to extract Cr(VI) from a liquid, the magnetic activated carbon adsorbent was created for the current investigation. H₃PO₄ was utilised as an activating agent in the thermal procedure to prepare activated carbon from marigold flower straw for the first time. The marigold flower straw activated carbon (MFSAC) was further decorated with Fe₃O₄ nanoparticles (Fe₃O₄/MFSAC) in the …

Virulence Analysis, Characterization Of Integron, And Antimicrobial Resistance Genes In Strains Of Acinetobacter Baumannii Recovered From The Hydrological Sub-Region Of Soto La Marina, Tamaulipas, Tunde Olarinde Olaniyan, Ana Verónica Martínez Vázquez, Susana Silvia Fernández Dávila, Cesar Marcial Escobedo Bonilla, Virgilio Bocanegra Garcia

Research Symposium

Background: Acinetobacter baumannii (A. baumannii) is an important bacterium due to its frequent recovery as a multidrug-resistant pathogen in hospital settings. Its antimicrobial resistance capacities made it an important challenge to health care. Colistin has been adopted as the last resort drug for managing infections associated with A. baumannii and many other multidrug-resistant Gram-negative bacteria (MDR-GNB). Because of the rise in the use of colistin for the treatment of MDR-GNB, the evolution and spread of colistin-resistant bacteria is possible thus raising a potential threat to public health. However, we have chosen to focus on A. baumannii, a …

Stepwise Regression Analysis To Assess Various Factors Affecting On Fresh Sperm Quality In Ouled Djellal Ram, Yamina Belkhiri, Souheyla Benbia, Farida Bouzebda-Afri, Zoubir Bouzebda, Ramzi Lamraoui

The Thai Journal of Veterinary Medicine

The aim of this study was to evaluate the effect of various factors on fresh semen quality in Ouled Djellal rams. Specifically, we examined the impact of semen collection method, season, ram age, testosterone levels, body weight , and testicular weight. Twelve mature Ouled Djellal rams were used in the trial. Six rams were trained for semen collection using an artificial vagina (AV), while semen from the other six rams was collected using an electroejaculator (EE) biweekly throughout the year. Stepwise regression analysis showed that 62.50% of the variation in sperm volume, 48.60% of the variation in mass motility, 62.70% …

Novel Nanoformulation Of Piperlongumine Attenuates Pancreatic Tumor Desmoplasia And Modulates Tumor Immune Responses, Vivek K. Kashyap, Reinaldo Mandes, Neeraj Chauhan, Daniel Zubieta, Mohammed Sikander, Eswara N.H.K Ghali, Bilal B. Hafeez, Murali M. Yallapu, Subhash C. Chauhan

Research Symposium

Pancreatic cancer (PanCa) is characterized by a lack of early diagnosis, poor response to available therapeutic modalities, and chemoresistance. Gemcitabine (GEM) is currently considered the most effective therapy for PanCa; however, it shows only a marginal survival benefit of 6 months. This poor drug response has been attributed to desmoplasia, causes suboptimal drug delivery, alters tumor microenvironment (TME), which includes tumor surrounding blood vessels, fibroblasts, immune cells, extracellular matrix, and other signaling molecules and induces chemo-resistance in tumors. To overcome these existing issues associated with chemotherapy, the identification and development of novel therapeutic modalities are a pressing need. Piperlongumine (PL) …

Thermal Carbonization Of Biomass Wood Dust And Algae Wastes Via Microwave-Assisted H3Po4: Desirability Function And Statistical Optimization For Methylene Blue Dye Removal, Raja Razuan Raja Derisa, Hazierul F. Awang, Muhammad Hakim Azman, Intan Nur Hidayah Abdul Rashid, Ahmad Hapiz, Ruihong Wu, Abdallah Reghioua, Zaher Mundher Yaseen, Lee D. Wilson

AUIQ Complementary Biological System

This research utilized a carbonization procedure via microwave irradiation assisted by H₃PO₄ to generate a cost-effective adsorbent (CWDAG) from wood dust (WD) and algal (AG) biomass. The resulting CWDAG adsorbent was characterized for its methylene blue (MB) dye adsorption properties. The activation process employs 800 W microwave radiation for 15 min under a nitrogen gas (99.99%) atmosphere. Multiple techniques were employed to study the physicochemical properties of CWDAG, such as FTIR, XRD, FSEM-EDX, pH_pzc, and BET. Box-Behnken design (BBD) was employed to optimize the three important parameters of adsorption, as follows: A: CWDAG dosage …

Biological Infections Associated With Wearing Cosmetic Contact Lenses: Risks, Pathogens, And Preventative Measures: Mini Review, Lina M. Shaker, Wan Nor Roslam Wan Isahak

AUIQ Complementary Biological System

Cosmetic contact lenses are increasingly favored worldwide for their ability to enhance or alter aesthetic appearance. However, their use carries a higher risk of ocular infections, particularly when proper hygiene practices are neglected. This narrative review explores the biological and clinical aspects of infections associated with cosmetic lens usage, with a focus on pathogens such as Pseudomonas aeruginosa, Staphylococcus aureus (S. aureus), and Acanthamoeba. Key factors influencing infection susceptibility include microbial adhesion, biofilm formation, reduced oxygen permeability, and the effects of cosmetic pigments. The review further evaluates the role of lens material properties, including advancements in antimicrobial materials, …

Methylene Blue Dye Removal From Aqueous Solution Using Activated Carbon Prepared From Corn Cob Stem: Kinetics, Isotherms And Mechanism Studies, Gopi Narasimha Murthy, Uttam Kumar Sahu

AUIQ Complementary Biological System

In this study, a waste-to-wealth work has been proposed where activated carbon was synthesized from waste corn cob stems and used for methylene blue (MB) dye removal. Corn cob stems activated carbon (CCSAC) had been prepared in microwave technique using H₃PO₄ as an activating agent where the impregnation ratio of corn cob stems to H₃PO₄ was 1:4. Then CCSAC characterized by FTIR, BET, XRD, SEM, EDS, and Raman spectroscopy. The CCSAC showed the mesoporous with a good surface area of 54.61 m²/g. Around 150 mg adsorbent dose, 120 min contact time, pH …

Assessing The Biological Impacts And Health Risks Of Uv Radiation In The Context Of Sustainable Development Goals: Review, Ruaa H. Abbas, Lina M. Shaker

AUIQ Complementary Biological System

Ultraviolet (UV) radiation, while essential for certain biological processes, presents significant risks to human health and ecosystems. Increased UV exposure, driven by ozone depletion and climate change, heightens threats such as DNA damage, skin cancer, immune suppression, and eye disorders, directly aligning with Sustainable Development Goal (SDG) 3: Good Health and Well-being. Additionally, UV radiation disrupts marine and terrestrial ecosystems, impacting biodiversity and productivity, relevant to SDGs 14 and 15. This review evaluates the health and environmental impacts of UV radiation and highlights recent advancements in UV monitoring technology. Key findings reveal substantial evidence linking UV exposure to accelerated skin …

Biocomposite Adsorbent Of Grafted Chitosan-Benzaldehyde/Lactobacillus Casei Bacteria For Removal Of Acid Red 88 Dye: Box-Benken Design Optimization And Mechanism Approach, Hasan M. Aghaa, Salis A. Musa, Ahmad Hapiz, Ruihong Wu, Khansaa Al-Essa, Ali Mohammed Saleh, Abdallah Reghioua

AUIQ Complementary Biological System

Herein, biohybrid biosorbent material was produced from grafting chitosan benzaldehyde/Lactobacillus casei (CHS-BZ/LAC) for the removal of anionic dye acid red 88 (AR88). CHS-BZ/LAC was fabricated using a hydrothermal process under the condition of 100 °C in an oven for 6 h. The optimization of AR88 dye adsorption by CHS-BZ/LAC was conducted by employing Box Bahnken design (BBD). The code and value of parameters used in this study were CHS-BZ/LAC dosage (A: 0.02-0.1 g/L), dye solution pH (B: 4-10), and contact time (C: 10-40 min). The BBD design shows that the highest removal of …

Using Network Pharmacology To Systematically Deduce The Molecular Mechanism Of The Selective Perinucleolar Compartment Inhibitor Ml246, Bhuvnesh P. Sharma, Himanshu N. Singh, Bhupesh Singh, Deepak Parashar, Kuldeep K. Roy, Murali M. Yallapu, Subhash C. Chauhan, Vivek K. Kashyap

Research Symposium

Metarrestin (ML246) is an orally bioavailable synthetic molecule that selectively disrupts the perinucleolar compartment (PNC) structure, and it has showed promise in preclinical studies for metastatic cancer therapy. However, the precise molecular mechanism of ML246 remains poorly understood. We investigated the topological and protein interaction network (PIN) analyses of ML246 to determine the molecular mechanism of ML246. To determine the regulatory effect of ML246 on the rewired PIN of ML246, an intercom was constructed using 25 oncogenic key proteins. These proteins were selected using the reverse pharmacophore matching approach (based on a fit score > 0.502) using the STRING database and …

First-In-Class Peptide Molecules Targeting The Mien1 Cancer Signaling Pathway, For Which No Inhibitors Have Been Identified To Date., Amit Tripathi, Jamboor K. Vishwanatha

Research Symposium

Background: Migration and Invasion Enhancer 1 (MIEN1) is a critical oncogene that drives cancer cell migration, invasion, and epithelial-to-mesenchymal transition (EMT) in various cancers. MIEN1 overexpression is linked to aggressive tumor progression and poor prognosis. Despite its established role in cancer biology, targeted therapeutic options remain unexplored. This study aimed to identify and characterize first-in-class therapeutic molecules targeting the MIEN1 signaling pathway to inhibit cancer progression.

Methods: To develop novel bio-active peptides, we leveraged conserved immunoreceptor tyrosine-based activation motif (ITAM) and prenylation motifs within the MIEN1 protein sequence. Using rational peptide design, we identified and synthesized bioactive peptides, namely LA3IK …