Novel Osteoporosis Therapeutic Targets Derived from Medical Biotechnology

en 38605738 Novel Osteoporosis Therapeutic Targets Derived from Medical Biotechnology Osteoporosis is known as the most prevalent metabolic bone disease. Population aging and increasing life span are making osteoporosis one of the most challenging age-related diseases 1. Based on reports in 2009, over 500 million people are struggling with this disease. Also, 1 in 5 men and 1 in 3 women over 50 years suffer from osteoporotic fractures during their lifetime. 1.5 million osteoporotic fractures are occurred annually in the United States and it cost the healthcare system in this country about $57 billion in 2018. The disease is caused by an imbalance in bone homeostasis, which is induced by bone formation not compensating for bone resorption. This process lowers the Bone Mineral Density (BMD) and makes bone susceptible to fracture 2,3. The significant burden of osteoporosis can be decreased since the disease is treatable. Based on the mechanisms resulting in osteoporosis, two main conventional drug categories are used for the treatment of the disease including anti-resorptive agents and anabolic agents 2. Conventional treatments consist of Selective Estrogen-Receptor Modulators (SERMs), bisphosphonates, parathyroid hormone analogs, and calcitonin. The need for more advanced therapies with fewer adverse effects arises from complications such as increasing risk of cardiovascular diseases, gastrointestinal problems, altering blood calcium levels, potential risks for women in estrogen-related therapies, etc. 4,5. As mentioned previously, the imbalance of bone metabolism causes osteoporosis. In this dynamic complex process of metabolism which is known as bone remodeling, connection or disruption of each specific intra-cellular or inter-cellular link can cause or treat the disease 6. Hence, this fact represents the remarkable role of medical biotechnology in osteoporosis treatment. Medical biotechnology as an upcoming main pillar of health-related science, has grown so fast in the field of treating disease as well as prevention and diagnosis using a variety of novel approaches 7. In recent years, biotechnology has introduced some novel therapeutic targets to the medical world to provide more efficient and safe therapies followed by diminishing osteoporosis as a burdensome disease. Monoclonal RANK Ligand (RANKL) antibodies and monoclonal sclerostin antibodies including denosumab, romosozumab, and blosozumab as recently rendered treatments to medicine, have achieved better therapeutic outcomes than conventional treatments 2. One of the main novel therapeutic targets is microRNA (miRNA)-based treatment. Modified nucleoside oligomers are the most common miRNA inhibitors used for new approaches. Since the clinical transformation of miRNA inhibitors is less difficult than lentivirus transfection, using them to affect the progress of osteoporosis is more feasible. Cathepsin K inhibitors which are involved in bone resorption in addition to remodeling, have been studied in many trials and are being investigated to find a promising one 8. Another innovation is stem cell therapy which is performed with cell sources from Embryonic Stem Cells (ESCs), induced Pluripotent Stem Cells (iPSCs), and Adult Stem Cells (ASCs). Stem cell therapy provides tissue regeneration which in osteoporosis translates to bone formation. Inducing the growth and differentiation of osteoblasts alongside decreasing activity of osteoclasts via cellular crosstalk are the main roles of stem cell therapy 4. The Wnt signaling pathway components are recently presented as one of the therapeutic targets. So many clinical trials are being conducted for each component to benefit from the bone mass-maintaining ability of these molecules 9. The sphingosine-1-phos-phate (S1P) signaling pathway and leucine zipper motif (APPL1) extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway and the Bone Morphogenetic Protein (BMP) signaling pathway are other targets to point out that play roles in osteogenesis, conducting osteogenic differentiation of stem cells, and promoting angiogenesis 5. A novel RANKL i-body nominated as ADR3 was introduced with features such as a high affinity for binding to human RANKL (hRANKL) and an ability to tolerate many different physical environments 3. Also, integrins as cell-adhesion transmembrane molecules have improved postmenopausal women’s BMD and made bone loss reversed 6. Furthermore, in recent years light has been shed on the bone-related roles of melatonin. Some of its important effects are bone biological rhythm regulatory effects, bone microenvironment modulation, and osteoporosis treatment 10. Population aging is greatly increasing osteoporosis prevalence and burden thus promoting osteoporosis to become an immense concern for the healthcare system. Hence, researching more effective therapies with fewer side effects has become urgent. Although studies introduced innovative targets as effective therapies, more investigations such as high-quality clinical trials are necessary to provide more evidence proving their efficiency. Despite all the mentioned conventional treatments, innovative targets, and upcoming therapeutic strategies, studying and developing molecular targets with more accurate and detailed mechanisms is indispensable. Undoubtedly, cooperation between basic sciences, especially neuroscience and biotechnology, and internal medicine can create a brighter future for the treatment of endocrine diseases 11-14. Antibodies, Bone density, Cathepsin K, MicroRNAs, Monoclonal, Osteogenesis, RANK Ligand, Stem cells, Wnt signaling pathway 1 2 https://www.ajmb.org/En/Article.aspx?id=60559 https://www.ajmb.org/PDF/En/FullText/60559.pdf SepidehHajivalizadeh School of Medicine, Tehran University of Medical Sciences, Tehran, Iran92112ShahinAkhondzadeh739

en 38605744 One-step and Rapid Identification of SARS-CoV-2 using Real-Time Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Background: SARS-CoV-2 as the cause of novel coronavirus disease (COVID-19) is a member of the family Coronaviridea that has generated an emerging global health concern. Controlling and preventing the spread of the disease requires a simple, portable, and rapid diagnostic method. Today, a standard method for detecting SARS-CoV-2 is quantitative real-time reverse transcription PCR, which is time-consuming and needs an advanced device. The aim of this study was to evaluate a faster and more cost-effective field-based testing method at the point of risk. We utilized a one-step RT-LAMP assay and developed, for the first time, a simple and rapid screening detection assay targeting the Envelope (E) gene, using specific primers. Methods: For this, the total RNA was extracted from respiratory samples of COVID-19 infected patients and applied to one-step a RT-LAMP reaction. The LAMP products were visualized using green fluorescence (SYBR Green I). Sensitivity testing was conducted using different concentrations of the designed recombinant plasmid (TA-E) as positive control constructs. Additionally, selectivity testing was performed using the influenza H1N1 genome. Finally, the results were compared using with conventional real time RT-PCR. Results: It was shown that the RT-LAMP assay has a sensitivity of approximately 15 ng for the E gene of SARS-CoV-2 when using extracted total RNA. Additionally, a sensitivity of 112 pg was achieved when using an artificially prepared TA-E plasmid. Accordingly, for the detection of SARS-CoV-2 infection, the RT-LAMP had high sensitivity and specificity and also could be an alternative method for real-time RT-PCR. Conclusion: Overall, this method can be used as a portable, rapid, and easy method for detecting SARS-CoV-2 in the field and clinical laboratories. COVID-19, Detection, LAMP Assay, Real time PCR, SARS-CoV-2 3 8 https://www.ajmb.org/En/Article.aspx?id=60560 https://www.ajmb.org/PDF/En/FullText/60560.pdf MohammadShoushtariDepartment of Virology, Pasteur Institute of Iran, Tehran, Iran92073MehdiZeinoddini92079JavadFathi91902HaniKeshavarz AlikhaniDepartment of Regenerative Medicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran92115FatemehShiekhiFaculty of Chemistry and Chemical Engineering, Malek Ashtar University of Technology, Tehran, Iran92116

en 38605742 Recombinant Production of TP4-LYC1, A New Chimeric Peptide with Targeted Cytotoxicity to HeLa Cells Background: Tilapia Piscidin 4 (TP4) showed potential anti-tumor effects against various cancer cells. Lycosine-1 (LYC1), is another Antimicrobial Peptides (AMP) from spider venom with targeted penetration to cancer cells without any adverse effects on normal cells. The aim of this study was to produce a soluble recombinant fusion peptide in order to diminish the cytotoxicity of TP4 against normal cells. Methods: In order to express of TP4-LYC-1, TP4, and LYC1 in fusion to the inteins1/2 of pTWIN-1 vector, induction condition was optimized to earn soluble peptides. Auto-cleavage induction of inteins1/2 was performed based on IMPACT® manual and their effect on cell viability of HeLa and HUVEC cells was surveyed by MTT assay. Results: The best condition for accessing the most soluble peptide in fusion to the inteins was approximately similar for all three peptides (0.1 mM of IPTG, at 22°C). After the induction of self-cleavage of inteins, a band in 3, 3, and 6 kDa was observed on tricine-SDS-PAGE. The IC50 values of TP4-LYC1 and TP4 against HeLa cells were calculated as 0.83, and 2.75 µM, respectively. Conclusion: In the present study, a novel chimeric peptide, TP4-LYC1, was successfully produced. This fusion protein can act as a safe bio-molecule with potent cytotoxic effects against cancer cells, but the penetration ability and determination of cell death mechanism must be performed in order to have more precise view on the apoptosis induction of this recombinant peptide.  Cell survival, HeLa cells, Inteins, Spider venoms, Tilapia 9 15 https://www.ajmb.org/En/Article.aspx?id=60561 https://www.ajmb.org/PDF/En/FullText/60561.pdf HaniehMohammad PourDepartment of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran92117AliJahanian-NajafabadiDepartment of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran92118FatemehShafiee92119

en 38605741 Effect of Intra-ovarian Injection of Mesenchymal Stem Cells or its Conditioned Media on Repeated OPU-IVEP Outcomes in Jersey Heifers and Its Relationship with Follicular Fluid Inflammatory Markers Background: Repeated Ovum Pick Up (OPU) could have a detrimental effect on ovarian function, reducing In Vitro Embryo Production (IVEP). The present study examined the therapeutic effect of adipose–derived Mesenchymal Stem Cells (MSCs) or its Conditioned Medium (ConM) on ovarian trauma following repeated OPU. Resolvin E1 (RvE1) and Interleukin-12 (IL-12) were investigated as biomarkers. Methods: Jersey heifers (n=8) experienced 11 OPU sessions including 5 pre-treatment and 6 treatment sessions. Heifers received intra-ovarian administration of MSCs or ConM (right ovary) and Dulbecco’s Modified Phosphate Buffer Saline (DMPBS; left ovary) after OPU in sessions 5 and 8 and 2 weeks after session 11. The concentrations of RvE1 and IL-12 in follicular fluid was evaluated on sessions 1, 5, 6, 9, and 4 weeks after session 11. Following each OPU session, the IVEP parameters were recorded. Results: Intra-ovarian administration of MSCs, ConM, and DMPBS did not affect IVEP parameters (p>0.05). The concentration of IL-12 in follicular fluid increased at the last session of pre-treatment (Session 5; p<0.05) and remained elevated throughout the treatment period. There was no correlation between IL-12 and IVEP parameters (p>0.05). However, RvE1 remained relatively high during the pre-treatment and decreased toward the end of treatment period (p<0.05). This in turn was associated with decline in some IVEP parameters (p<0.05). Conclusion: Intra-ovarian administration of MSCs or ConM during repeated OPU did not enhance IVEP outcomes in Bos taurus heifers. The positive association between RvE1 and some of IVEP parameters could nominate RvE1 as a promising biomarker to predict IVEP parameters following repeated OPU. Animals, Biomarkers, Cattle, Conditioned medium, Follicular fluid, Inflammation, Interleukin-12, Ovary, Resolvin E1, Stem cells 16 28 https://www.ajmb.org/En/Article.aspx?id=60562 https://www.ajmb.org/PDF/En/FullText/60562.pdf AliSarvariReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran335AmirNiasari-NaslajiDepartment of Theriogenology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran92120AbolfazlShirazi82BanafshehHeidariDepartment of Photo Healing and Regeneration, Medical Laser Research Center, Yara Institute, ACECR, Tehran, Iran503SaraBorjian BoroujeniReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran337Mohammad HosseinMoradiDepartment of Animal Sciences, Faculty of Agriculture and Natural Resources, Arak University, Arak, Iran92121Mohammad MehdiNaderiReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran334BaharehBehzadiReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran506Mohammad-Mahdi MehrazarReproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran92122Mohammad MehdiDehghanDepartment of Surgery and Radiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran92123

en 38605740 Orexin-1 Receptor Antagonist SB-334867 Enhances Formalin-Induced Nociceptive Behaviors in Adult Male Rats Background: Orexin (hypocretin) is one of the hypothalamic neuropeptides that plays a critical role in some behaviors including feeding, sleep, arousal, reward processing, and drug addiction. Neurons that produce orexin are scattered mediolaterally within the Dorsomedial Hypothalamus (DMH) and the lateral hypothalamus. In the current research, we assessed the impact of prolonged application of the antagonist of Orexin Receptor 1 (OXR1) on nociceptive behaviors in adult male rats. Methods: Sixteen Wistar rats received subcutaneous (s.c.) injections of the OXR1 antagonist, SB-334867 (20 mg/kg, i.p.), or its vehicle repetitively from Postnatal Day 1 (PND1)-PND30. On the 30th day following the final application of the OXR1 antagonist formalin-provoked pain was evaluated by injecting formalin. Results: Administration of the OXR1 antagonist in the long-term augmented the formalin-provoked nociceptive behaviors in interphase and phase II of the formalin-induced pain. Conclusion: Current results showed that the continued inhibiting OXR1 might be implicated in formalin-induced nociceptive behaviors. Therefore, the present study highlighted the effect of orexin on analgesia. Formalin, Nociception, Orexin receptor 1, Orexin, SB-334867 29 33 https://www.ajmb.org/En/Article.aspx?id=60563 https://www.ajmb.org/PDF/En/FullText/60563.pdf MasoumehKourosh-Arami92125AlirezaKomakiDepartment of Neuroscience, School of Science and Advanced Technologies in Medicine, Hamadan University of Medical Sciences, Hamadan, Iran92126MasoumehGholamiDepartment of Physiology, Medical College, Arak University of Medical Sciences, Arak, Iran92127

en 38605743 Annexin-A5 Overexpression Increases Sensitivity of MCF-7 and MCF-7/ADR Cells to Epirubicin Background: Multi-drug resistance is an important challenge in the chemotherapy of cancer. The role of annexin A5 (ANXA5) in the biology of cancer has been the focus of many studies. Breast Cancer (BC) is frequent cancer in women with high morbidity and mortality rate. The present study aimed to investigate the effects of ANXA5 overexpression on the anti-tumor activity of Epirubicin (EPI) in MCF-7 and MCF-7/ADR cells. Methods: MCF-7 and MCF-7/ADR cells were transfected with the pAdenoVator-CMV-ANXA5-IRES-GFP plasmid or mock plasmid. The overexpression of ANXA5 was evaluated using qPCR. The effects of ANXA5 overexpression and EPI on the cell viability of MCF-7 and MCF-7/ADR cells were measured using an MTT assay. Cell apoptosis was measured by annexin V/7-AAD flow cytometry assay. Results: Following the overexpression of ANXA5, the viability of MCF-7 and MCF-7/ADR was significantly decreased. Furthermore, the overexpression of ANXA5 in MCF-7 cells increased the cytotoxic effects of EPI in all doses and reduced the IC50 of EPI from 17.69 µM to 4.07 µM. Similarly, the overexpression of ANXA5 in MCF7-ADR cells reduced the IC50 of EPI from 27.3 µM to 6.69 µM. ANXA5 overexpression alone or combined with EPI treatment increased the apoptosis of MCF7 and MCF7-ADR cells.  Conclusion: The results of the present study demonstrate that ANXA5 overexpression increases the sensitivity of MCF-7 and MCF-7/ADR to EPI, suggesting a possible beneficial role of ANXA5 in the therapy of BC. Annexin-A5, Antineoplastic agents, Breast neoplasms, Drug resistance, MCF7 cells 34 39 https://www.ajmb.org/En/Article.aspx?id=60564 https://www.ajmb.org/PDF/En/FullText/60564.pdf MahshadGhasemiDivision of Medical Biotechnology, Department of Laboratory Sciences, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran92128NiloofarReiazi Division of Medical Biotechnology, Department of Laboratory Sciences, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran92129AbbasBehzad-BehbahaniDiagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran899Mohammad AliTakhshidDiagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran986

en 38605737 Extracellular L-Asparaginase Synthesis Bacillus niacin Isolation, Optimization, and Characterization from Marine Saltern Sediment Sources Background: Asparagine is an amino acid that can be converted into aspartic acid and ammonia by the enzyme L-asparaginase. Some forms of cancer, such Acute Lymphoblastic Leukaemia (ALL) and Non-Hodgkin Lymphoma (NHL), respond well to this enzyme when employed as a chemotherapeutic drug. The purpose of this research was to find bacteria that can manufacture the enzymes L-asparaginasein marine slattern sediment which can be employed in commercial and industrial scale production. Methods: All of the strains were identified as Bacillus niacini spp. by biochemical and molecular testing. The strain belongs to the Bacillus genus, according to nutritional, biochemical, PCR and 16srRNA sequencing data. Results: According to the findings of this research, Bacillus niacin spp. have the potential to create a substance that is helpful in a variety of medical applications. The results of this study hint to the possibility that bacteria have the ability to produce antimicrobial compounds, which have the potential to be successful in a wide variety of environments. Conclusion: Numerous opportunities may arise for researchers interested in utilizing the medical potential of enzyme-producing bacteria if they are successfully isolated and screened from aquatic and terrestrial habitats. Asparagine, Asparaginase, Bacillus niacini, Polymerase chain reaction 40 48 https://www.ajmb.org/En/Article.aspx?id=60565 https://www.ajmb.org/PDF/En/FullText/60565.pdf Mugip RahamanAbdul Wahab Department of Biotechnology, Dr. M.G.R Educational and Research Institute, Chennai, Tamil Nadu, Iran92131ThirunavukkarasuPalaniyandi Department of Anatomy, Biomedical Research Unit and Laboratory Animal Centre, Saveetha Dental College and Hospital, Saveetha Institute of Medical and Technical Science, Saveetha University, Chennai, Tamil Nadu, India92132JohnWyson Department of Food Processing Technology, AMET University, Kanathur, Chennai, Tamil Nadu, India92133AshaSivajiDepartment of Biochemistry, DKM College for Women, Vellore-632001, Tamil Nadu, India92134SwarnakalaThamadaMolecular Systematics Laboratory, Zoological Survey of India, Andaman & Nicobar Regional Centre, Port Blair - 744 102, Andaman and Nicobar Islands92135

en 38605736 Anti-Quorum Sensing and Anti-Biofilm Activity of Ginger (Zingiber officinale) Rhizomes against Multidrug-Resistant Clinical Isolates of Pseudomonas aeruginosa Background: The aim of this study was to determination of Anti-Quorum Sensing (AQS) and anti-biofilm potential of the methanol extract of ginger (Zingiber officinale) rhizomes against multidrug-resistant clinical isolates of Pseudomonas aeruginosa (P. aeruginosa). Methods: The AQS activity of ginger was determined against Chromobacterium violaceum (C. violaceum) ATCC 12472 (CV12472), a biosensor strain, in qualitative manner using the agar well diffusion method. The violacein pigment inhibition was assessed to confirm AQS activity of ginger. The AQS potential of sub-minimum Inhibitory Concentrations (sub-MICs) of the ginger extract was determined by targeting different QS regulated virulence factors, including swarming motility (using swarm diameter measurement method), pyocyanin pigment (using chloroform extraction method), Exopolysaccharide (EPS) (using phenol-sulphuric acid method), and biofilm formation (using microtiter plate assay), against clinical isolates (CIs 2, 3, and 4) and standard reference strain of P. aeruginosa (PA01). Results: The AQS activity of methanol extract of ginger was confirmed against C. violaceum (CV12472) as inhibition of violacein pigment formation without effecting the growth of CIs and PA01 of P. aeruginosa. The ginger extract exhibited concentration-dependent inhibition of virulence factors and biofilm formation. The maximum reduction was found in swarming motility, pyocyanin, EPS and biofilm formation against PA01 (51.38%), CI3 (57.91%), PA01 (63.29%) and CI2 (64.37%), respectively at 1/2 MIC of ginger extract. Conclusion: The results of present study revealed the effective AQS and anti-biofilm potential of Zingiber officinale rhizome methanol extract at a reduced dose (sub-MICs). The extract may be explored as an agent of antimicrobial compounds having AQS and anti-biofilm activity for controlling microbial infection and also for reducing the chances of emergence of resistance in P. aeruginosa.   Chromobacterium violaceum, Ginger, Pseudomonas aeruginosa, Pyocyanin, Rhizome 49 56 https://www.ajmb.org/En/Article.aspx?id=60566 https://www.ajmb.org/PDF/En/FullText/60566.pdf PankajKumar Sagar Department of Microbiology, Bundelkhand University, Jhansi-284128, Uttar Pradesh, India92136PoonamSharma Department of Zoology, Indira Gandhi National Tribal University (A Central University), Amarkantak-484886 , Madhya Pradesh, India92137RambirSingh92138

en 38605739 Green Tea Extract Reduced Lipopolysaccharide-Induced Inflammation in L2 Cells as Acute Respiratory Distress Syndrome Model Through Genes and Cytokine Pro-Inflammatory Background: Acute Respiratory Distress Syndrome (ARDS) is a severe lung inflammatory condition that has the capacity to impair gas exchange and lead to hypoxemia. This condition is found to have been one of the most prevalent in patients of COVID-19 with a more serious condition. Green tea (Camellia sinensis L.) contains polyphenols that possess many health benefits. The purpose of this study was to assess the anti-inflammatory activities of green tea extract in Lipopolysaccharide (LPS)-induced lung cells as ARDS cells model. Methods: In this study, rat lung cells (L2) were induced by LPS to mimic the inflammation observed in ARDS and later treated with green tea extract. Pro-inflammatory cytokines such as Interleukin (IL)-12, C-Reactive Protein (CRP) as well as Tumor Necrosis Factor-α (TNF-α) were investigated using the ELISA method. Gene expression of NOD-Like Receptor Protein 3 (NLRP-3), Receptor for Advanced Glycation End-product (RAGE), Toll-like Receptor-4 (TLR-4), and Nuclear Factor-kappa B (NF-κB) were evaluated by qRTPCR. Apoptotic cells were measured using flow cytometry. Results: The results showed that green tea extract treatment can reduce inflammation by suppressing gene expressions of NF-κB, NLRP-3, TLR-4, and RAGE, as well as pro-inflammatory cytokines such as IL-12, TNF-α, and CRP, an acute phase protein. Apoptosis levels of inflamed cells also found to be lowered when green tea extract was administered; thus, also increasing live cells compared to non-treated cells. Conclusion: These findings could lead to the future development of supplements from green tea to help alleviate ARDS symptoms, especially during critical moments such as the current pandemic. Acute respiratory distress syndrome, Camellia sinensis, Cytokines, Inflammation, Tea 57 65 https://www.ajmb.org/En/Article.aspx?id=60567 https://www.ajmb.org/PDF/En/FullText/60567.pdf DidikPriyandoko Biology Study Program, Faculty of Mathematics and Natural Sciences, Indonesia University of Education, Bandung 40154, Indonesia92139WahyuWidowati41601LennyLenny Faculty of Biotechnology, Atma Jaya Catholic University of Indonesia, BSD Campus, Tangerang 15345, Indonesia92141SintyaNoviantiFaculty of Biotechnology, Atma Jaya Catholic University of Indonesia, BSD Campus, Tangerang 15345, Indonesia92142RevikaRevikaFaculty of Biotechnology, Atma Jaya Catholic University of Indonesia, BSD Campus, Tangerang 15345, Indonesia92143HannaSari Widya KusumaBiomolecular and Biomedical Research Center, Aretha Medika Utama, Bandung 40163, Indonesia92144IkaAdhani SholihahSchool of Life Sciences and Technology, Bandung Institute of Technology, Bandung 40163, Indonesia41610