Biotechnology Institute Research Articles

Upregulation of Proinflammatory Variables of Analytical Importance and their Potential Interplay in the Pathogenesis of Rheumatoid Arthritis

Objective: Study role of the selected inflammatory markers and their importance in the development and progression of rheumatoid arthritis. Study Design: Cross-sectional study Place and Duration: Institute of Molecular Biology and Biotechnology, The University of Lahore, (11-months) Methodology: A total of 50 patients diagnosed with rheumatoid arthritis in the Department of Orthopedics (Allama Iqbal Medical College, Jinnah Hospital, Lahore-Pakistan) and 50 healthy individuals of the age of 35-45 were included into the study. Glutathione, Catalase, Maliondialdehyde, Superoxide dismutase, Glutathione peroxidase, Nitric oxide (NO) and vitamin C, E and A was estimated spectrophotometrically. Serum C-Reactive Protein, IL-21, TNF-α, AOPPs, AGE’s and MMP-3 was measured by ELISA method using Human-ELISA kits (Glory) Results: A statistically noteworthy raise of all the biomarkers level in serum of rheumatoid arthritis (RA) was seen (p=0.001) comparable with healthy individuals. The correlation between joint damage progression diagnostic biomarker AGE and MMP-3 levels (r=0.485, 95% p=0.001) was highly significant. CRP and IL-21 also showed highly significant raise in joint inflammation (r=0.359, p=0.001). The results significantly depicted that model of MMP-3 levels at the onset of the disease and CRP and IL-21 were the robust interpreters of the erosion development. Similarly, antioxidants including Vitamin E, Vitamin C and Vitamin A were decreased in levels of (p<0.011), (p<0.011) and (p<0.022) respectively in RA subjects than in control subjects. Vitamin E, C and A are the most imperative free radical foragers within membrane. They also act as the major line of resistance against free radicals. Conclusion: Development of pain in joints in rheumatoid arthritis condition is due to the increase in the oxidative stress which is the key contrivance in the pathogenesis of RA. Keyword: Reactive oxygen species, Oxidative stress, oxidation of lipid peroxides, Rheumatoid arthritis, Osteoarthritis, osteocyte damage.

Digital data transformation: optimizing CGT processes and accelerating scale-up & tech transfer for manufacturing success

Bridging the data gap between R&D and manufacturing with purpose-built softwareBest practices on using technology to ensure highly complex CGT processes are successfully replicated and controlledEstablishing reasonable expectations for data visibility and sharing in sponsor/CMO relationshipsIdentifying the right time to build a digital data strategy and key IT requirements when choosing a software platformTeodor has 14 years of expertise as a scientist in biotech process development, design, optimization, and validation. His successful track record building and managing technology roadmaps for GMP manufacturing, including software selection, qualification, and implementation, position him as a respected advisor on maximizing operational efficiency to accelerate scale-up, tech transfer, and commercial release. Before joining IDBS, Teo served as Process Development Scientist at Emergent BioSolutions, where he led science and technology for a newly developed orphan disease vaccine. Previously at Merck Healthcare, his roles spanned Bioprocess Development Scientist and Compliance Specialist for Downstream Processing. Teo holds a BS in Biomedical Engineering from Yale University and an MS in Biotechnology and Bioengineering from EPFL (Swiss Federal Institute of Bioengineering).

Pollock liver oil extraction using a new enzyme obtained from the Aspergillus oryzae strain

A distinctive feature of fish fats from fats of plants and terrestrial animals is the presence in their composition of highly unsaturated and polyunsaturated omega-3 fatty acids (eicosapentaenoic and docosahexaenoic acids), which causes their high biological value. A typical type of raw material for obtaining edible fat is cod liver, which makes up about 10% of the weight of fish and contains an average of about 50% lipids. The Far Eastern pollock liver with a high content of polyunsaturated fatty acids has similar characteristics. From a technological point of view, the complexity of extracting these biologically valuable components is due to the high sensitivity of fish oil lipids to oxidation. The article discusses the use of new enzyme Protorizin-LAP obtained at Russian scientific research institute of food biotechnology using Aspergillus oryzae strain - a producer of a complex of exo- and endopeptidases with an increased activity of leucine aminopeptidase. Oil yield (%) and acid value (mg KOH/g oil) are considered as efficiency criteria for enzymatic hydrolysis. Optimization results of enzymatic hydrolysis of the pollock liver using a full-factorial experiment for two factors are presented. The optimal values of the factors are established: the dosage of the enzyme preparation Protorizin-LAP (0.4% by weight of the raw material) and the duration of the enzymatic hydrolysis process (1 hour). The study of the comparative enzymatic analysis of pollock liver showed that the use of the enzyme preparation "Protorizin LAP" in the technological process of processing fat-containing fish. The laboratory sample is not inferior to commercial analogues in terms of the ability to extract fat from pollock liver and may be promising in the processing of other raw materials of animal origin.

Cocaine derivatives–synthesis, analytical characterization and evaluation of their cross reactivity in immunochemical assays

Synthetic cocaine derivatives are currently marketed on various internet websites as legal cocaine alternatives. These psychoactive substances include phenyltropanes (troparil, dichloropane, RTI-55), 1,4-dialkyl piperazines, phenyl piperidines, benztropines, and dimethocaine. Although they have been sold as “research chemicals” for more than a decade, they have not been studied systematically. Their consumption evades forensic-toxicological perception through the absence of specific identification methods. For a holistic forensic investigation of synthetic cocaine derivatives, the availability of reference compounds is a limiting factor. Moreover, synthetic side products, stereoisomers, degradation products, or deuterated references have not been isolated and characterized. Therefore, the immunochemical cross-reactivity of cocaine derivatives will be investigated after their synthesis and analytical characterization. Diverse phenyltropanes, e.g., troparil, dichloropane, and phenyl piperidines, were synthesized using documented methods. In addition, stereoisomers and deuterated references as well as metabolites were also produced. The reactions were carefully monitored, and all products were purified and characterized with NMR spectroscopy, high-resolution mass spectrometry, and polarimetry. In addition, the synthesized substances and their metabolites, which are structurally closely related to cocaine and benzoylecgonine, were tested in a CEDIA cocaine assay to evaluate their cross-reactivity. In cooperation with the Institute of Pharmacy and Molecular Biotechnology in Heidelberg, the aimed phenyltropanes and phenyl piperidines were successfully synthesized and analytically characterized. All synthesized compounds were tested in the CEDIA cocaine assay in concentrations of 100 and 1000 ng/mL. Altogether none of the compounds showed significant cross-reactivity to create a false-positive result. The availability of reference compounds is often a bottleneck of research in forensic chemistry and toxicology. Therefore, the synthesis and characterization of reference compounds such as cocaine derivatives can support the development of methods for their identification and quantification. In addition, investigating authorities can gain better insides into the quality of the sold substances and the production methods in clandestine labs. Due to their structural similarities with cocaine and benzoylecgonine, a cross-reactivity in immunochemical assays was presumed but can be excluded for the CEDIA cocaine assay. All aimed synthetic cocaine derivatives were successfully synthesized and characterized. These substances will support the development of analytical methods and support their coverage in the daily forensic routine. The tested cocaine derivatives showed no significant cross-reactivity in the CEDIA cocaine immunoassay. While immunochemical assays are very helpful for the initial identification of drug users by police and forensic-toxicological laboratories, the cross-reactivity should be evaluated in other test systems.

The RSNA ImageShare — Enabling Secure, Transparent, and Easy Image Exchange

In the early 2000s, the radiology community was awakened to the limitations of electronic media (CDs, DVDs) for exchanging imaging exams. Clinicians frustrated by the time-consuming task of opening discs, while Internet-based exchange of music, photos, and videos were becoming more widespread. The RSNA, which had extensive experience working on interoperability issues in medical imaging, began to look for opportunities to address the issue. In 2007, in the wake of the financial crisis, the National Institute of Biomedical Imaging and Bioengineering (NIBIB) issued an RFP to address Internet-based exchange of medical images. The RFP defined requirements for the network, including that it needed to be patient controlled and standards based. The RSNA was awarded funding for what came to be known as RSNA ImageShare. Over the next 8 years, the RSNA worked in partnership with several vendors and academic institutions to create a network for sharing image-enabled personal health records (PHR). The foundation of interoperability standards used in ImageShare was provided by Integrating the Healthcare Enterprise (IHE), a standards-development organization with which RSNA has had a long association. In 2018 and 2019, the RSNA looked at what had been accomplished and asked if we could take that next step at a national level and promote a solution by which any standards-compliant party could exchange imaging exams through an HIE mechanism.

Technology valuation for intellectual property commercialization

This study was conducted to evaluate the technical value of 19 intellectual property rights held by the Korea Research Institute of Bioscience and Biotechnology, and the purpose of this valuation is to promote commercialization by acquiring the technology transfer or exclusive license for the intellectual property rights. This study examines the theoretical and practical aspects of the valuation methods and procedures of the income approach that are useful for valuing intellectual property. Assuming that bioscience and biotechnology firms receive license transfer of intellectual property held by a foreign company, the valuation analysis of intellectual property based on the profit approach is based on business plans and financial statements of domestic companies. After calculating the operating profit from the gross profit of the company in detail, the corporation tax and the capital cost are taken into account and the depreciation cost is increased or decreased to calculate the excess profit and multiply the present value by the present value. We propose an income approach model and a case analysis to obtain the ultimate value of intellectual property by multiplying the contribution by this factor. It is not easy to predict future cash flows and estimate various financial statements, and there is a limit to the possibility that the evaluator will be subject to the estimation of the appropriate discount rate. More detailed and partial complementary research classified by the industry size is to be left as a future study.

Effect of Organic and Inorganic Fertilizers on Growth, Yield & Quality of Cucumber (Cucumis sativus L.) under Protected Cultivation

The present investigation entitled “Effect of organic and inorganic fertilizer on growth, yield & quality of cucumber (Cucumis Sativus L.) Under protected cultivation” was carried out in the Polyhouse of Jacob Institute of Biotechnology and Bioengineering, Sam Higginbottom University of Agriculture, Technology and Sciences, Prayagraj, Uttar Pradesh, during winter season from September to December 2021.The experiment was laid out in Randomized complete Block Design comprising of 10 treatments with three replications. The study revealed that the treatment T10(Recommended Dose of Fertilizer (RDF) 50% + Poultry manure 50%) performed best in terms of no. of leaves (107.62/plant),days to first flowering(36.94), days to harvest (62.33), fruit length (15.03 cm), fruit diameters (14.1 mm),number of fruits per plant (20.70),weight (162.40 g), yield per plot (3.26 Kg), yield per 200 sq.m (571.52 Kg) and Vitamin C (7.57 mg/100 g fresh fruit). However, treatment T7 (25% RDF + 75% FYM) has maximum number days to fruit set (57.36) and treatment T4(RDF 50% + FYM 50%) had highest soluble solid (3.77oBrix). The significantly higher gross return (Rs 17,145.6/ha), Net Profit (Rs 13,415.21/ha) and B:C ratio (3.59:1) was also recorded under treatment T10 ((RDF) 50% + Poultry manure 50%). Overall results revealed that T10 (RDF) 50% + Poultry manure 50%) was found to be the most suitable over all the other treatments in relation to growth and yield of Cucumber.

Biocatalysis: Improving Enzymatic Processes through Protein and Reaction Engineering

ADVERTISEMENT RETURN TO ISSUEEditorialNEXTBiocatalysis: Improving Enzymatic Processes through Protein and Reaction EngineeringCarlos A. Martinez*Carlos A. MartinezBiocatalysis Center of Excellence (BCE) and Chemical R&D, Pharmaceutical Sciences, Small Molecules, Pfizer Inc., Groton Laboratories, Eastern Point Road, Groton, Connecticut 06340, United States*Phone: 860-5018743. Email: [email protected]More by Carlos A. Martinez, Nicholas J. TurnerNicholas J. TurnerDepartment of Chemistry and Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K.More by Nicholas J. Turnerhttps://orcid.org/0000-0002-8708-0781, and Zhi LiZhi LiDepartment of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, Singapore 117585More by Zhi Lihttps://orcid.org/0000-0001-7370-2562Cite this: Org. Process Res. Dev. 2022, 26, 7, 1855–1856Publication Date (Web):July 15, 2022Publication History Published online15 July 2022Published inissue 15 July 2022https://doi.org/10.1021/acs.oprd.2c00179Copyright © Published 2022 by American Chemical SocietyRIGHTS & PERMISSIONSArticle Views1239Altmetric-Citations-LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (482 KB) Get e-AlertsSUBJECTS:Alcohols,Amines,Biocatalysis,Peptides and proteins,Reaction engineering Get e-Alerts

Safety Assessment of a Fungal-based Red Colorant Produced by Monascus purpureus MTCC 25436

Pigment production by Monascus species for application to a wide array of food products has been explored in many countries. However, possible contamination with citrinin, a mycotoxin, has been a concern with different Monascus-based products leading to a regulated usage of the microbial food additive worldwide. In the Philippines, there is no known local large-scale producer of microbial pigments. A red colorant from Monascus purpureus MTCC 25436 was successfully produced and patented by the National Institute of Molecular Biology and Biotechnology–University of the Philippines Los Baños (NIMBB-UPLB). The produced colorant was subjected to various safety assessment tests to determine its safety for food applications. The AMES test determined that the product was not capable of inducing reverse mutations in Salmonella typhimurium mutant strains, and this non-mutagenicity of the colorant is further confirmed by the mammalian micronucleus test. Establishing the LD50 was done to determine the lethal dose of the colorant. No model animal died, even after feeding with the highest allowable experimental concentration. Based on the dermal sensitization test, the colorant was a very weak sensitizer. Lastly, detection of citrinin was done using high-performance liquid chromatography. According to the generated chromatograms, citrinin was at non-detectable levels. In conclusion, the colorant obtained from the local strain M. purpureus MTCC 25436 is safe for consumption. The results of the study can be used by the local regulating agencies as a baseline for possibly crafting regulations for the registration of microbial-derived food colorants, which the Philippines currently lacks.

Оценка исходного материала для получения риса с высоким содержанием амилозы

Rice is one of the main crops of strategic importance. Rice consumption in Kazakhstan is increasing compared to other cereals and reaches more than 65 %. Today, due to the increase in imports of rice cereals, it seems relevant to develop rice production, increase its competitiveness, and create domestic rice varieties with improved nutritional and culinary qualities. A key factor in determining the quality of rice is the percentage of amylose in the starch of the rice grain. In this regard, this work is devoted to the search for donors of the trait “high amylose content” among varieties and lines of domestic and foreign breeding in the rice collection of the RSE on the REM “Institute of Plant Biology and Biotechnology” (RSE on the REM “IPBB”). Screening of 172 foreign and domestic varieties and lines of rice for amylose content made it possible to identify 9 high-amylose variety samples. Among them one domestic ancient variety of Kazakh selection K3077 Kazakhi was found, which can serve as a donor of the trait “high amylose content”. Because of the climatic conditions of Kazakhstan and the long growing season, only six foreign varieties and lines were selected: Mavr (26.8 %), Taibonet (25.4 %), Parity (27, 0 %), K3612 Kara-Kyltyk (25.0 %), K3077 Kazakhs (25.1 %), and Solnechny (25.4 %). The selected varieties and lines of rice with a high content of amylose will be used as donors of the “high amylose content” trait for rice breeding.

P-796 Intraovarian injection of plasma rich in growth factors improves ovarian reserve and reproductive outcomes in women with diminished ovarian reserve

Abstract Study question Does Plasma rich in growth factors (PRGF) ovarian injection increase the ovarian reserve biomarkers in women with diminished ovarian reserve? Summary answer PRGF injection increased Antimüllerian hormone (AMH) levels and antral follicular count (AFC) since the first follow-up after treatment allowing the initiation of ovarian stimulation cycles. What is known already Several techniques based on in vitro activation, ovarian fragmentation and stem cell ovarian transplantation have been proposed to reactivate ovarian function and increase IVF success in women with premature ovarian insufficiency and poor ovarian response. However, less invasive and feasible approaches are still required for those patients where egg donation is the only practical option. Platelet rich plasma (PRP) and PRGF contain a high concentration of platelets, which carry more than 800 types of proteins, cytokines, hormones, and chemoattractants. Indeed, intraovarian PRP injection has recently been used in different case series and cohort studies of POI women with encouraging results. Study design, size, duration Retrospective study with 104 women aging 23-45 years who received a PRGF intraovarian injection (REGENERA-Ovario) was conducted between 2020 and 2021 at IVIRMA Alicante (Spain). Study was approved by the IRB committee of La Fe University Hospital (2112-FIVI-109-SH). Participants/materials, setting, methods Patients underwent a PRGF injection (Endoret kit; B.T.I. Biotechnology Institute S.L, Spain) in both ovaries and a follow up of ovarian reserve biomarkers (AFC, AMH) and follicle stimulating hormone (FSH) to evaluate follicular reactivation seeking IVF or spontaneous pregnancy. The follow up visits ranged from 1 to 5 months and were developed on a monthly basis after treatment or after menses recovery in POI. Study variables were compared to basal levels with a paired t-test. Main results and the role of chance Overall, 104 women (age: 38.7±2.0 years; BMI: 22.1±2.9) with diminished ovarian reserves underwent a PRGF intraovarian injection (3.3±0.8ml PRGF/each ovary). Prior to treatment, our cohort was characterized high serum FSH (21.8±4.1mIU/mL), low AMH levels (0.25±0.37ng/ml), and an ovarian volume of 6.3±2.3cm3. Eighty-eight of them fulfilled the 1st follow up visit and showed an increase of serum AMH (0.25±0.37ng/ml vs. 0.32±0.36ng/ml, p = 0.008), and AFC in both ovaries (2.1±1.9 vs. 3.2±2.4, p < 0.0001) compared to basal levels, achieving ovarian reactivation in 55.9% of them and IVF cycle initiation in 19.3%. These improvements were higher 2 months after treatment (55 patients), with increased antral follicles (AFC-2: 2.9±2.5, p = 0.0001) and ovarian activation in 58.2%, and IVF start in 43.9% of patients. Similar results were observed during the 3rd follow up accomplished by 30 women and lasted until the 5th visit (n = 8), although the reduced patient number at this point. A total of 44 IVF cycles were started and oocyte pick-up was successfully developed in 72.7% of them with a mean number of 3.3±4.2MII oocytes, 1.8±2.1 fertilized oocytes and 1.5±2.0 embryos per cycle. Indeed, 6 pregnancies were obtained during this period, 2 of them after embryo transfer and 4 by natural conception. Limitations, reasons for caution Our results are encouraging but a large cohort of patients with a longer follow up period is needed to establish the efficacy and duration of the PRGF positive ovarian effects. Moreover, a proper comparison with a control group with the same characteristics and no PRGF intervention is still required. Wider implications of the findings PRGF intraovarian injection reactivates follicle growth and allows IVF cycle initiation and embryo generation in a poor prognosis population of patients with diminished ovarian reserve. The effects persisted for several months after treatment. Trial registration number 2112-FIVI-109-SH

Modeling interaction between non-structural protein 2 of Chikungunya Virus and various protein factors of innate pathway

Chikungunya virus is positive-sense single-stranded RNA virus that causes an arthropod-borne chikungunya fever, myalgia and arthralgia. Chikungunya virus belongs to the Togaviridae family, and the genus is Alphavirus. Virus-host protein interaction plays a vital role in developing vaccines and antiviral drugs. We designed the current study to establish the in-silico interaction of non-structural protein 2 (nsP2) with proteins of innate immune pathway. The nsP2 sequences of various Chikungunya virus genotypes were retrieved from National Centre for Biotechnology Institute (NCBI). The homology models of proteins were generated through a protein modeling online web server. Protein-protein interaction (PPI) between nsP2 and proteins of innate immune pathway were docked using High Ambiguity-Driven Docking (HADDOCK) webserver. The interactive residues of the bimolecular complexes were analyzed with PDBsum-Generate online webserver. Our findings revealed differentially affinity of nsP2 of various chikungunya genotypes towards key proteins of cellular innate pathway. The nsP2 of Asian genotype demonstrates relatively high interaction with interferon-beta promoter stimulator 1 (IPS-1). Similarly, nsP2 of various genotypes binds with differential affinity to tumor necrosis factor receptor-associated factor 6 (TRAF6) with the highest affinity observed for the nsP2 of the West African genotype. Bimolecular complexes of nsP2 and host proteins demonstrate the interaction of various domains of nsP2 with proteins of the innate immune pathway. Thus, it is sought that the selected panel of the proteins might be helpful to treat the viral infection as a therapeutic drug target in the future.

Whole genome re-sequencing of indian wheat genotypes for identification of genomic variants for grain iron and zinc content.

Whole-genome sequencing information which is of abundant significance for genetic evolution, and breeding of crops. Wheat (Triticum spp) is most widely grown and consumed crops globally. Micronutrients are very essential for healthy development of human being and their sufficient consumption in diet is essential for various metabolic functions. Biofortification of wheat grains with iron (Fe) and zinc (Zn)has proved the most reliable and effective way to combat micronutrient associated deficiency. Genetic variability for grain micronutrient could provide insight to dissect the traits. In the current study, 1300 wheat lines were screened for grain Fe and Zn content, out of which only five important Indian wheat genotypes were selected on the basis of Fe and Zn contents. These lines were multiplied during at the National Agri-Food Biotechnology Institute (NABI) and re-sequenced to identify genomic variants in candidate genes for Fe and Zn between the genotypes. Whole genome sequencing generated ̴ 12 Gb clean data. Comparative genome analysis identified 254 genomic variants in the candidate genes associated with deleterious effect on protein function. The present study demonstrated the fundamental in understanding the genomic variations for Fe and Zn enrichment to generate healthier wheat grains.

Abstract 2695: Transcriptomic profiling of human patients with acute myeloid leukemia reveals critical effects of growth hormone responsiveness on patient survival and mechanisms of tumor therapy resistance

Abstract Growth hormone (GH) has been reported to drive leukemia cell proliferation as early as 1977, while downstream effectors of the GH signaling pathway (i.e., insulin-like growth factor 1; IGF1) have more recently been widely implicated in the aggressive pathology and robust therapy resistance phenotypes of acute myeloid leukemia (AML). It is also known that patients with acromegaly (high levels of circulating GH and IGF1) develop AML at a significantly higher rate than the general population, while individuals with GH resistance remain completely resistant to all types of cancer. Despite all of the aforementioned evidence pointing to a critical role of GH in AML progression and prognosis, a detailed transcriptomic analysis of human patients is lacking. We aim to address this gap by thorough in silico studies aimed at revealing the nature and extent to which the GH axis can be suitable targeted in AML. Here, we present a summary of our analyses of clinical and transcriptomic data from 200 human patients with AML from The Cancer Genome Atlas database, using multiple bioinformatics software programs in the public domain. Our study revealed that GH receptor (GHR) is expressed more highly in AML than in normal myeloid cells, and a robust correlation exists between GH responsiveness and patient survival. A sex-specific profiling of the patient transcriptomic data identified critical pathways related to therapy resistance, extracellular matrix remodeling, metastasis, and immune evasion to be upregulated differentially in male and female AML patients with relatively elevated GHR expression compared to patients with relatively lower GHR expression. Furthermore, we identified microRNAs which are differentially expressed in AML patients with higher than median GHR expression. We validated these miRNAs to be effective indicators of disease progression and prognosis and can be valuable markers in liquid biopsy approaches. Acknowledgement: This work was supported in part by the State of Ohio’s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, and Ohio University’s Edison Biotechnology Institute. Citation Format: Emily Davis, Reetobrata Basu, John J. Kopchick. Transcriptomic profiling of human patients with acute myeloid leukemia reveals critical effects of growth hormone responsiveness on patient survival and mechanisms of tumor therapy resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2695.

Abstract 2620: Combination of growth hormone receptor antagonist and gemcitabine leads to highly efficacious tumor clearance in a mouse model of pancreatic ductal adenocarcinoma

Abstract Pancreatic cancer is responsible for more than 48,000 deaths and 60,000 new cancer cases every year in the United States while the overall 5-year survival rate is only 10%. These numbers point towards an urgent need for new therapeutic approaches wherein therapy resistance poses a tremendous challenge. We and others have identified the role of growth hormone (GH) and its cognate receptor (GHR), overexpressed in pancreatic ductal adenocarcinoma patients (PDAC; >90% of pancreatic cancer cases), in driving therapy resistance by directly upregulating multidrug efflux, phenotype switch via epithelial-to-mesenchymal transition (EMT) and inhibition of apoptosis. Our laboratory had pioneered the discovery of pharmacologic GHR antagonists including the only approved and prescribed GHR antagonist, pegvisomant, and a new antagonist candidate named compound-G. Here, we used in vitro, in vivo and in silico analyses to assess the feasibility and efficacy of a treatment regimen including GHR antagonist and chemotherapy to manage PDAC. Both pegvisomant and compound-G exhibit significant suppression of cancer cellular viability, invasive potential, drug efflux rate and apoptosis in PDAC cell cultures. In male and female Nude mice with human PDAC xenografts, GHR inhibition by either pegvisomant or compound-G markedly re-sensitized the tumors towards both low and high doses of gemcitabine treatment. Compound G plus high doses of gemcitabine lead to almost complete tumor clearance in 40% of the mice. Also, tissue analyses reveal increased production of GH in tumors treated with chemotherapy. In the GHR antagonist treated tumors, significantly lower expression of ABC transporter (Abcb1, Abcg5, Abcc2, Abcg2), EMT mediator (Cdh2, Zeb1, Snai2, Epas1) and apoptosis marker (Bcl2) genes compared to control were observed - accounting for the observed sensitization to gemcitabine. In the human PDAC patient datasets, increased expression of GHR correlates strongly with increased expression of multidrug efflux transporters, anti-apoptotic markers, and EMT transcription factors. Therefore, we provide robust pre-clinical validation for a treatment regimen combining GHR antagonist and chemotherapy leading to highly efficacious tumor clearance. Acknowledgement: This work was supported in part by the State of Ohio’s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, and Ohio University’s Student Enhancement Award and Edison Biotechnology Institute. Citation Format: Reetobrata Basu, Prateek Kulkarni, Emily Davis, Silvana Duran, John J. Kopchick. Combination of growth hormone receptor antagonist and gemcitabine leads to highly efficacious tumor clearance in a mouse model of pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2620.

Abstract 1004: Growth hormone upregulates tumor-derived exosomal distribution of EMT mediators and multi-drug efflux pumps in melanoma

Abstract Melanoma is the most aggressive form of skin cancer with a dismal 5-year survival rate of 16% at the metastatic stage of the disease. Exosomes have recently emerged as a critical mediator of cancer therapy resistance by regulating epithelial to mesenchymal transition (EMT) and drug efflux. Exosomes are extracellular vesicles (diameters 30-150nm) released from almost all types of cells including tumor cells which package and transfer proteins, RNA, and lipids (often called cargo) from one cell to another. This intercellular communication via exosomes has been reported to induce a drug-resistant phenotype across the tumor by transporting ABC multidrug transporters and mediators of EMT. Multiple studies have established that pituitary-, or tumor- or tumor microenvironment-derived growth hormone (GH) promotes EMT and drug resistance in cancer by binding to GHR-expressing tumor cells. A question arises: Does GH exert this effect across the tumor population via modulating tumor-derived exosomal cargo? No study has yet directly assessed the effect of GH on exosomes in terms of modulating EMT or drug resistance. We isolated exosomes from human melanoma cells with differential levels of GHR expression, following treatment with combinations of GH, anti-cancer drugs, and the GHR antagonist, pegvisomant. We observed no change in the quantity of tumor-derived exosomes with changes in GH action. However, we found significant upregulation of multidrug efflux pumps ABCB1, ABCC1, ABCC2, and ABCG2 in GH-treated tumor exosomes. Importantly, treatment of naïve melanoma cells with these exosomes increased their drug efflux rate and reduced drug retention. Exosomes derived from pegvisomant treated melanoma cells showed markedly reduced ABC transporter contents and did not increase the efflux rate of naïve tumor cells. A similar trend in matrix metalloproteinases (MMP2, MMP9) and TGFb levels in isolated exosomes as well as TGFb signaling and invasion capacity in naïve tumor cells treated with isolated exosomes were observed. These results present a unique effect of GH on tumor-derived exosomal content modulation towards propagating a drug-resistant tumor phenotype in naïve cancer cells. Acknowledgment: This work was supported in part by the State of Ohio’s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, and Ohio University’s Student Enhancement Award and Edison Biotechnology Institute. Citation Format: Prateek Kulkarni, Reetobrata Basu, John J. Kopchick. Growth hormone upregulates tumor-derived exosomal distribution of EMT mediators and multi-drug efflux pumps in melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1004.

The Interaction of Biotechnology and Institution: A Stakeholder Perspective

Institutional systems have a crucial impact on the development of biotechnology. In this article, we analyze the interaction between biotechnology and institutions. To conduct our analysis, we use the case study method and the stakeholder perspective. Our findings suggest the following: (1) Through the analysis of patent data, biotechnology has been developing very rapidly in recent years in China; (2) basic biotechnology institutions have been established, consisting of government, policy, and other institutional arrangements; (3) the interaction between the development of biotechnology and its existing institutions is dynamic; and (4) the interaction is affected by relative stakeholders. This study contributes to the theory concerning the governance of biotechnology, which is important in the sustainable development of biotechnology. Moreover, the article sheds light on policy implications.

Optimizing Plant Growth-Promoting Rhizobacteria to Improve the Relationship Between Above and Below Ground Biodiversity

Biodiversity plays a crucial role in ensuring environmental sustainability. Plantrhizosphere microbiome interactions are ecologically important because they can alter the biological, chemical, and physical properties of the soil, ultimately affecting plant productivity. To maintain soil function, bio-ameliorant technologies, new modern composting crop residues, and limited use of synthetic pesticides have all been used to boost microbial activity in the soil. Soil management biotechnology is the use of microbes to maintain soil function, resulting in increased plant productivity. It offers a novel way to deal with a wide range of issues that are currently unsolved by conventional technology. It is thought that an improved relationship between above and below ground biodiversity can contribute to the re-establishment of essential biological functions. The Indonesian Research Institute for Biotechnology and Bioindustry (IRIBB), PT Riset Perkebunan Nusantara, has been working on developing plant growth-promoting rhizobacteria (PGPR) as a bio-ameliorant to improve the relationship between above and below ground biodiversity for sustainable management and increased plant productivity.
 Keywords: bio-ameliorant, biodiversity, biotechnology, rhizosphere, soil microbes

ПАМ’ЯТІ ВИДАТНОГО УЧЕНОГО-БІОЛОГА, ПРОФЕСОРА, ЧЛЕН-КОРЕСПОНДЕНТА НАН УКРАЇНИ, ЗАСЛУЖЕНОГО ДІЯЧА НАУКИ І ТЕХНІКИ ГРИГОРЮКА ІВАНА ПАНАСОВИЧА

On May 19, 2022, the heart of the famous biologist, Doctor of Biological Sciences, Professor, and corresponding member of the NAN of Ukraine, Honored Science and Technology Officer Hryhoriuk Ivan Panasovych stopped beating. 
 Ivan Panasovych was born on October 24, 1941, in the village of Toporivtsi of the Novoselytsia district, Chernivtsi region. After graduating from school, he entered the Faculty of Biology of the University of Chernivtsi, graduating in 1965 with a degree in Plant Physiology and qualified as a biologist, plant physiologist, and teacher of biology and chemistry. He was doing his postgraduate studies from 1966 to 1969 at the Institute of Plant Physiology of the Academy of Sciences of the USSR (now the Institute of Plant Physiology and Genetics of the National Academy of Sciences of Ukraine), where he was employed until 2005. He was the Candidate in Biological Sciences since 1978, Doctor of Biological Sciences since 1996, Professor since 2000. 
 In 2005 Ivan Panasovych, Professor of Ecobiotechnology and Biodiversity, was appointed the Director of the NNC of Biotechnology, Biotech Service, and Bioenergy Conversion of the National Agricultural University. In 2007 he headed the Educational and Scientific Institute for Nature Conservation and Biotechnology and at the same time worked as a professor at the Department of Plant Physiology, Ecology and Biomonitoring of the National University of Bioresources and Nature Management of Ukraine (NUBIP). 
 He is one of the founders of a scientific school in Ukraine for the study of theoretical and experimental physiology of regulatory systems of water exchange, physiological and molecular biological mechanisms of stability, and adaptation of cultivated plants to environmental stressors.
 
 
 Corresponding Member of the NAN of Ukraine I. P. Hryhoriuk – author and co-author of more than 1000 scientific, educational and popular scientific works, including 35 monographs, 10 books, and brochures, 5 textbooks, 3 dictionaries, 630 articles (50 of them – abroad), 34 certificates and patents for inventions and 68 scientific and practical recommendations and guidelines, most of which are introduced into production. He was the scientific director of 12 candidate and a scientific consultant for 5 doctoral dissertations.
 For his conscientious work I. P. Hryhoriuk was awarded many national awards. On June 5, 2009, by the Decree of the President of Ukraine I. P. Hryhoriuk was awarded the honorary title «Honored figure of science and technology of Ukraine» for a significant personal contribution to solving environmental problems and preservation of the environment. He was a winner of the State Prize of Ukraine in the field of science and technology in 2011.

AI recognition of patient race in medical imaging: a modelling study

Previous studies in medical imaging have shown disparate abilities of artificial intelligence (AI) to detect a person's race, yet there is no known correlation for race on medical imaging that would be obvious to human experts when interpreting the images. We aimed to conduct a comprehensive evaluation of the ability of AI to recognise a patient's racial identity from medical images. Using private (Emory CXR, Emory Chest CT, Emory Cervical Spine, and Emory Mammogram) and public (MIMIC-CXR, CheXpert, National Lung Cancer Screening Trial, RSNA Pulmonary Embolism CT, and Digital Hand Atlas) datasets, we evaluated, first, performance quantification of deep learning models in detecting race from medical images, including the ability of these models to generalise to external environments and across multiple imaging modalities. Second, we assessed possible confounding of anatomic and phenotypic population features by assessing the ability of these hypothesised confounders to detect race in isolation using regression models, and by re-evaluating the deep learning models by testing them on datasets stratified by these hypothesised confounding variables. Last, by exploring the effect of image corruptions on model performance, we investigated the underlying mechanism by which AI models can recognise race. In our study, we show that standard AI deep learning models can be trained to predict race from medical images with high performance across multiple imaging modalities, which was sustained under external validation conditions (x-ray imaging [area under the receiver operating characteristics curve (AUC) range 0·91-0·99], CT chest imaging [0·87-0·96], and mammography [0·81]). We also showed that this detection is not due to proxies or imaging-related surrogate covariates for race (eg, performance of possible confounders: body-mass index [AUC 0·55], disease distribution [0·61], and breast density [0·61]). Finally, we provide evidence to show that the ability of AI deep learning models persisted over all anatomical regions and frequency spectrums of the images, suggesting the efforts to control this behaviour when it is undesirable will be challenging and demand further study. The results from our study emphasise that the ability of AI deep learning models to predict self-reported race is itself not the issue of importance. However, our finding that AI can accurately predict self-reported race, even from corrupted, cropped, and noised medical images, often when clinical experts cannot, creates an enormous risk for all model deployments in medical imaging. National Institute of Biomedical Imaging and Bioengineering, MIDRC grant of National Institutes of Health, US National Science Foundation, National Library of Medicine of the National Institutes of Health, and Taiwan Ministry of Science and Technology.