1.Neutralization of SARS-CoV-2 spike pseudotyped viruses using postbiotic preparations from Lactiplantibacillus plantarum BS25.
Joshua Angelo Hermida Mandanas ; Day Yu-Chao ; Leslie Michelle M. Dalmacio
Acta Medica Philippina 2026;60(7):58-75
BACKGROUND
Despite the presence of SARS-CoV-2 antivirals, namely, remdesivir, nirmatrelvir and toremifene, these drugs entail adverse effects and limited effectiveness. Thus, development of a safer alternative is imperative, and likely candidates include the cell-free supernatant (CFS) and protein fractions from Lactiplantibacillus plantarum strains. These postbiotics have known antiviral properties primarily mediated by plantaricins and enhanced by organic acids.
OBJECTIVESThe study determined the in silico mechanism of plantaricins against the receptor-binding domain (RBD) and angiotensin-converting enzyme 2 (ACE2), as well as compared the in vitro activities of the L. plantarum BS25 CFS, semi-purified and crude protein fractions, against the SARS-CoV-2 pseudotyped viruses (nCoV-S-EGFP) expressing the spike of wild-type (wt) Wuhan strain, Omicron BA.1 and BA.4/5 variants. Further, this study determined the metabolome of the strain BS25 CFS.
METHODSA quasi-experimental approach was utilized in the study. Plantaricins were screened for safety in silico, followed by molecular docking with RBD and ACE2. Using a cell culture model, two-fold dilutions of the CFS and fractions were tested for cytotoxicity and microneutralization against the pseudoviruses. Mass spectrometry was utilized for the metabolomics.
RESULTSPlantaricins interact stably with RBD than ACE2 which were mediated by hydrogen, hydrophobic and covalent bonds. The activities of the CFS and fractions were substantially higher against the nCoV-SEGFP BA.4/5 variant. The 1:8 dilution of CFS entailed no cytotoxicity and displayed higher activities than toremifene. Metabolomics has identified a relatively abundant putative peptide, followed by organic acids and cyclic peptides.
CONCLUSIONPlantaricins can prevent SARS-CoV-2 entry by interacting with key RBD mutations or with intrinsically disordered RBD residues. The cationic and hydrophobic RBD mutations in the BA.4/5 variant may facilitate interactions with the putative peptide from the CFS and fractions, hence the observed potent activities. These findings can be used as basis for the development of an alternative antiviral targeting the RBD of live or pseudotyped SARS-CoV-2 variants.
Sars-cov-2
2.Knowledge, Attitude, and Practices Assessment of Unvaccinated Adult In-patients Regarding COVID-19 Vaccination in a Tertiary Hospital in Nueva Vizcaya, Philippines: A Cross-sectional Study.
Charity May B. Pazziwagan ; Emmeline B. Borillo
Acta Medica Philippina 2026;60(3):78-87
OBJECTIVES
his quantitative cross-sectional study aimed to determine the level of knowledge, attitude, and practices
regarding COVID-19 vaccination of unvaccinated adult patients admitted at Region II Trauma and Medical Center
(R2TMC). This may help determine the knowledge gap regarding COVID-19 vaccination, address it, and to achieve
the goal to vaccinate all eligible Filipinos. Furthermore, this may also be a springboard for future researches and
management regarding novel infections needing new vaccinations.
Collected data from the sample size of 197 using the formula for finite population with 95% confidence
level, population proportion of 50%, population size of 400, allowing 5% margin of error were summarized in a
Microsoft excel database and tables, then were analyzed using Jamovi version 2.6.22 software. Frequency, mean,
and standard deviation were calculated to measure the knowledge, attitude, and practices; Pearson correlation and
Kendall’s Tau b correlation for its relationship.
There is moderate level of knowledge (MS: 9.80-11.8), a positive attitude (MR: 2.35-2.60), and good preventive
practices (MS: 12.00-13.00) regarding SARS-CoV-2 (COVID-19) vaccination among the unvaccinated adult in-patients
of R2TMC post-pandemic. Knowledge, attitudes, and practices statistically differed (pCONCLUSION
There is a significant correlation among the knowledge of the respondents about COVID-19 vaccination,
their attitude towards it, and their practices, implying that promoting preventive behaviors toward COVID-19 would
require promoting both knowledge and efficacy beliefs among the public.
Human ; Bacteria ; Male ; Female ; Adult: 25-44 Yrs Old ; Middle Aged: 45-64 Yrs Old ; Wounds And Injuries ; Population Density ; Cross-sectional Studies ; Forecasting ; Sars-cov-2 ; Sars Virus ; Covid-19
3.External ocular manifestations among patients diagnosed with Coronavirus disease 2019 in a referral center in the Philippines.
Alyssa Louise B. Pejana-Paulino ; Aramis B. Torrefranca Jr. ; Nilo Vincent DG. Florcruz ; Ma. Dominga B. Padilla
Acta Medica Philippina 2026;60(1):69-77
BACKGROUND AND OBJECTIVES
The global pandemic caused by Coronavirus Disease 2019 (COVID-19) has affected millions, with growing evidence of the potential role of ocular tissues in viral transmission. At the time of writing, local data regarding the phenomenon was limited. This study investigated external ocular manifestations in patients with COVID-19 at a referral center in the Philippines, examined correlations between demographics, systemic manifestations, and laboratory results with ocular manifestations, and determined their timing relative to systemic symptoms.
METHODSThis single-center, descriptive cross-sectional study was carried out from December 8 to 18, 2020 at the adult COVID-19 wards of the Philippine General Hospital involving 72 participants. Data collection involved relevant clinical history taking and performing gross eye examination. The prevalence of ocular manifestations was described with 95% confidence intervals. Correlations between ocular manifestations and quantitative variables were analyzed with point-biserial correlation, and associations with qualitative variables were tested using chi-square or Fisher’s exact tests.
RESULTSAmong participants, 31.9% presented with ocular manifestations with foreign body sensation as the most prevalent ocular symptom (11.1%) and conjunctival hyperemia as the most prevalent ocular finding (19.4%). The median age of patients with ocular manifestations was 41 years old with a higher prevalence in the male population (73.9%, CI=95%, p=0.001). No significant correlation was observed between presence of external ocular manifestations and the different systemic and ocular co-morbidities as well as with COVID-19 clinical classification. Among those who experienced symptoms, majority (29.2%) of the patients experienced systemic symptoms prior to the onset of ocular symptoms. Ocular complaints may present as the sole manifestation (13.9%). Several laboratory parameters were measured and only temperature and AST levels showed a low positive correlation with the presence of ocular manifestations.
CONCLUSIONOcular manifestations occur in roughly one third of patients with COVID-19 based on this study population. With some individuals presenting with ocular signs or symptoms as the initial and sole manifestation, healthcare practitioners must exercise caution and remain vigilant in managing patients who present as such. At the time of writing, this is the first local study investigating the different external ocular manifestations in patients with COVID-19. There is a need to pursue more robust studies and conduct more local investigations which will guide both ophthalmologists and other practitioners in strengthening existing guidelines regarding precautionary practices, clinical diagnosis, and management of COVID-19 patients.
Human ; Sars-cov-2 ; Covid-19 ; Philippines ; Adult ; Association ; Classification ; Collection ; Confidence Intervals ; Coronavirus ; Cross-sectional Studies ; Data Collection ; Demography ; Diagnosis ; Disease ; Exercise ; Eye ; Foreign Bodies ; History ; Hospitals ; Hospitals, General ; Hyperemia ; Laboratories ; Male ; Morbidity ; Ophthalmologists ; Pandemics ; Patients ; Population ; Prevalence ; Referral And Consultation ; Role ; Sensation ; Temperature ; Time ; Tissues ; Volition ; World Health Organization ; Writing
4.Genomic variant surveillance of SARS-CoV-2 positive specimens using a direct PCR product sequencing surveillance (DPPSS) method.
Nicole Ann L. Tuberon ; Francisco M. Heralde III ; Catherine C. Reportoso ; Arturo L. Gaitano III ; Wilmar Jun O. Elopre ; Kim Claudette J. Fernandez
Acta Medica Philippina 2026;60(1):57-68
BACKGROUND AND OBJECTIVE
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as the causative agent of COVID-19 has significantly challenged the public health landscape in late 2019. After almost 3 years of the first ever SARS-CoV-2 case, the World Health Organization (WHO) declared the end of this global health emergency in May 2023. Although, despite the subsequent drop of COVID-19 cases, the SARS-CoV-2 infection still exhibited multiple waves of infection, primarily attributed to the appearance of new variants. Five of these variants have been classified as Variants of Concern (VOC): Alpha, Beta, Gamma, Delta, and the most recent, Omicron. Therefore, the development of methods for the timely and accurate detection of viral variants remains fundamental, ensuring an ongoing and effective response to the disease. This study aims to evaluate the feasibility of the application of an in-house approach in genomic surveillance for the detection of SARS-CoV-2 variants using in silico designed primers.
METHODSThe primers used for the study were particularly designed based on conserved regions of certain genes in the virus, targeting distinct mutations found in known variants of SARS-CoV-2. Viral RNA extracts from nasopharyngeal samples (n=14) were subjected to quantitative and qualitative tests (Nanodrop and AGE). Selected samples were then analyzed by RT-PCR and amplicons were submitted for sequencing. Sequence alignment analysis was carried out to identify the prevailing COVID-19 variant present in the sample population.
RESULTSThe study findings demonstrated that the in-house method was able to successfully amplify conserved sequences (spike, envelope, membrane, ORF1ab) and enabled identification of the circulating SARS-CoV-2 variant among the samples. Majority of the samples were identified as Omicron variant. Three out of four designed primers effectively bound into the conserved sequence of target genes present in the sample, revealing the specific SARSCoV-2 variant. The detected mutations characterized for Omicron found in the identified lineages included K417N, S477N, and P681H which were also identified as mutations of interest. Furthermore, identification of the B.1.448 lineage which was not classified in any known variant also provided the potential of the developed in-house method in detecting unknown variants of COVID-19.
CONCLUSIONAmong the five VOCs, Omicron is the most prevalent and dominant variant. The in-house direct PCR product sequencing surveillance (DPPSS) method provided an alternative platform for SAR-CoV-2 variant analysis which is accessible and affordable than the conventional diagnostic surveillance methods and the whole genome sequencing. Further evaluation and improvements on the oligonucleotide primers may offer significant contribution to the development of a specific and direct PCRbased detection of new emerging COVID-19 variants.
Sars-cov-2 ; Polymerase Chain Reaction ; Dna Primers ; Oligonucleotide Primers ; Computer Simulation ; Conserved Sequence ; Coronavirus ; Covid-19 ; Disease ; Emergencies ; Evaluation Studies As Topic ; Genes ; Genome ; Global Health ; Health ; Identification (psychology) ; Infection ; Infections ; Membranes ; Methods ; Mutation ; Oligonucleotides ; Organizations ; Population ; Public Health ; Rna ; Rna, Viral ; Sars Virus ; Sequence Alignment ; Severe Acute Respiratory Syndrome ; Syndrome ; Viruses ; Whole Genome Sequencing ; World Health Organization
5.National Trends in Influenza Vaccination Rates in South Korea Before and During the COVID-19 Pandemic, 2011-2022.
Kyeongeun KIM ; Kyeongmin LEE ; Yejun SON ; Seoyoung PARK ; Raphael UDEH ; Jiseung KANG ; Hayeon LEE ; Soeun KIM ; Jaeyu PARK ; Hyeon Jin KIM ; Damiano PIZZOL ; Lee SMITH ; Jiyoung HWANG ; Dong Keon YON
Biomedical and Environmental Sciences 2025;38(9):1044-1057
OBJECTIVE:
Despite the global decrease in influenza infections during the coronavirus disease 2019 (COVID-19) pandemic, seasonal influenza remains a significant health issue. South Korea, known for its robust pandemic response and high influenza vaccination rates, offers a unique context for examining changes in vaccination trends during the pandemic. Using nationally representative data, we aimed to understand the impact of the pandemic on influenza vaccination behavior over a 12-year period and to identify vulnerable groups.
METHODS:
We analyzed influenza vaccination rates in South Korea between 2011-2022, focusing on pandemic-related impacts. The data of 2,426,139 adults (≥ 19 years) from the Korea Community Health Survey were used to assess demographic and sociological factors influencing vaccination behaviors.
RESULTS:
We observed an increase in influenza vaccination rates during the pre-COVID-19 period from 2011-2013 (weighted prevalence: 46.68% [95% confidence interval ( CI): 46.55-46.82]) to 2017-2019 (weighted prevalence: 52.50% [95% CI: 52.38-52.63]). However, a significant decline was observed in 2022, the late-COVID-19 pandemic period (weighted prevalence: 55.78% [95% CI: 55.56-56.01]), compared with the mid-pandemic period in 2021 (weighted prevalence: 59.12% [95% CI: 58.91-59.32]), particularly among populations traditionally prioritized for influenza vaccination, including older adults (≥ 65 years) and patients with chronic diseases and low educational and income levels.
CONCLUSION
The influenza vaccination rate in South Korea was significantly affected by the COVID-19 pandemic, showing a notable decrease among vulnerable demographic groups. This suggests the need for targeted public health strategies to address vaccine hesitancy and improve vaccination rates, particularly among high-risk populations.
Humans
;
Republic of Korea/epidemiology*
;
COVID-19/epidemiology*
;
Adult
;
Middle Aged
;
Influenza Vaccines/administration & dosage*
;
Male
;
Female
;
Influenza, Human/epidemiology*
;
Aged
;
Vaccination/statistics & numerical data*
;
Young Adult
;
Pandemics
;
SARS-CoV-2
6.Pathogenicity and Transcriptomic Profiling Revealed Activation of Apoptosis and Pyroptosis in Brain of Mice Infected with the Beta Variant of SARS-CoV-2.
Han LI ; Bao Ying HUANG ; Gao Qian ZHANG ; Fei YE ; Li ZHAO ; Wei Bang HUO ; Zhong Xian ZHANG ; Wen WANG ; Wen Ling WANG ; Xiao Ling SHEN ; Chang Cheng WU ; Wen Jie TAN
Biomedical and Environmental Sciences 2025;38(9):1082-1094
OBJECTIVE:
Patients with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection frequently develop central nervous system damage, yet the mechanisms driving this pathology remain unclear. This study investigated the primary pathways and key factors underlying brain tissue damage induced by the SARS-CoV-2 beta variant (lineage B.1.351).
METHODS:
K18-hACE2 and C57BL/6 mice were intranasally infected with the SARS-CoV-2 beta variant. Viral replication, pathological phenotypes, and brain transcriptomes were analyzed. Gene Ontology (GO) analysis was performed to identify altered pathways. Expression changes of host genes were verified using reverse transcription-quantitative polymerase chain reaction and Western blot.
RESULTS:
Pathological alterations were observed in the lungs of both mouse strains. However, only K18-hACE2 mice exhibited elevated viral RNA loads and infectious titers in the brain at 3 days post-infection, accompanied by neuropathological injury and weight loss. GO analysis of infected K18-hACE2 brain tissue revealed significant dysregulation of genes associated with innate immunity and antiviral defense responses, including type I interferons, pro-inflammatory cytokines, Toll-like receptor signaling components, and interferon-stimulated genes. Neuroinflammation was evident, alongside activation of apoptotic and pyroptotic pathways. Furthermore, altered neural cell marker expression suggested viral-induced neuroglial activation, resulting in caspase 4 and lipocalin 2 release and disruption of neuronal molecular networks.
CONCLUSION
These findings elucidate mechanisms of neuropathogenicity associated with the SARS-CoV-2 beta variant and highlight therapeutic targets to mitigate COVID-19-related neurological dysfunction.
Animals
;
COVID-19/genetics*
;
Mice
;
Brain/metabolism*
;
Apoptosis
;
Mice, Inbred C57BL
;
SARS-CoV-2/physiology*
;
Pyroptosis
;
Gene Expression Profiling
;
Transcriptome
;
Male
;
Female
7.Does Prenatal SARS-CoV-2 Infection Exacerbate Postpartum Lower Urinary Tract Symptoms? A Multicenter Retrospective Cohort Study.
Yu Han LYU ; Min LI ; Hui Qing YAO ; Tian Zi GAI ; Lin LIANG ; Su PAN ; Ping Ping LI ; Ya Xin LIANG ; Yue YU ; Xiao Mei WU ; Min LI
Biomedical and Environmental Sciences 2025;38(9):1095-1104
OBJECTIVE:
Coronavirus disease 2019 (COVID-19) can result in fatigue and post-exertional malaise; however, whether severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection exacerbates lower urinary tract symptoms (LUTS) is unclear. This study investigated the association between prenatal SARS-CoV-2 infection and postpartum LUTS.
METHODS:
A multicenter, retrospective cohort study was conducted at two tertiary hospitals in China from November 1, 2022, to November 1, 2023. Participants were classified into infected and uninfected groups based on SARS-CoV-2 antigen results. LUTS prevalence and severity were assessed using self-reported symptoms and the Incontinence Impact Questionnaire-Short Form (IIQ-7). Pelvic floor muscle activity was measured using electromyography following the Glazer protocol. Group comparisons were performed to evaluate the association of SARS-CoV-2 infection with LUTS and electromyography parameters, with stratified analyses conducted using SPSS version 26.0.
RESULTS:
Among 3,652 participants (681 infected, 2,971 uninfected), no significant differences in LUTS prevalence or IIQ-7 scores were observed. However, SARS-CoV-2 infection was an independent factor influencing the electromyographic activity of the pelvic floor muscles (mean tonic contraction amplitudes), regardless of delivery mode ( P = 0.001).
CONCLUSION
Prenatal SARS-CoV-2 infection was not significantly associated with an increased risk of postpartum LUTS but independently altered pelvic floor muscle electromyographic activity, suggesting potential neuromuscular effects.
Humans
;
Female
;
COVID-19/epidemiology*
;
Retrospective Studies
;
Adult
;
Pregnancy
;
Lower Urinary Tract Symptoms/virology*
;
Postpartum Period
;
Pregnancy Complications, Infectious/virology*
;
China/epidemiology*
;
Electromyography
;
SARS-CoV-2/physiology*
;
Pelvic Floor/physiopathology*
;
Prevalence
8.Progress on Wastewater-based Epidemiology in China: Implementation Challenges and Opportunities in Public Health.
Qiu da ZHENG ; Xia Lu LIN ; Ying Sheng HE ; Zhe WANG ; Peng DU ; Xi Qing LI ; Yuan REN ; De Gao WANG ; Lu Hong WEN ; Ze Yang ZHAO ; Jianfa GAO ; Phong K THAI
Biomedical and Environmental Sciences 2025;38(11):1354-1358
Wastewater-based epidemiology has emerged as a transformative surveillance tool for estimating substance consumption and monitoring disease prevalence, particularly during the COVID-19 pandemic. It enables the population-level monitoring of illicit drug use, pathogen prevalence, and environmental pollutant exposure. In this perspective, we summarize the key challenges specific to the Chinese context: (1) Sampling inconsistencies, necessitating standardized 24-hour composite protocols with high-frequency autosamplers (≤ 15 min/event) to improve the representativeness of samples; (2) Biomarker validation, requiring rigorous assessment of excretion profiles and in-sewer stability; (3) Analytical method disparities, demanding inter-laboratory proficiency testing and the development of automated pretreatment instruments; (4) Catchment population dynamics, reducing estimation uncertainties through mobile phone data, flow-based models, or hydrochemical parameters; and (5) Ethical and data management concerns, including privacy risks for small communities, mitigated through data de-identification and tiered reporting platforms. To address these challenges, we propose an integrated framework that features adaptive sampling networks, multi-scale wastewater sample banks, biomarker databases with multidimensional metadata, and intelligent data dashboards. In summary, wastewater-based epidemiology offers unparalleled scalability for equitable health surveillance and can improve the health of the entire population by providing timely and objective information to guide the development of targeted policies.
China/epidemiology*
;
Humans
;
Wastewater/analysis*
;
COVID-19/epidemiology*
;
Public Health
;
Wastewater-Based Epidemiological Monitoring
;
SARS-CoV-2
9.Effectiveness of chlorhexidine (CHX) mouthwash in the reduction of SARS-COV-2 viral load among COVID-19 patients: A systematic review
Aldren R. Remon ; Lady Jane R. Malagum ; Lovie Japhet S. Lopez
Philippine Journal of Nursing 2025;95(2):119-128
BACKGROUND
Public health was again at risk due to the appearance of new COVID-19 variants. Coronavirus Disease (COVID19) transmission occurs mainly through the oral mucosa. Specifically, nurses who care for or interact with hospitalized patients directly have a higher risk of contracting the virus. In order to manage the viral load in respiratory secretions, nurses need to be aware of evidence-based practices as clinical care providers. The unsophisticated application and the effectiveness of Chlorhexidine (CHX) Mouthwash as an independent nursing intervention to mitigate cross-contamination and reduce SARSCoV-2 transmission remains clinically significant.
OBJECTIVEThe study's objective was to determine the effectiveness of Chlorhexidine (CHX) mouthwash in reducing SARSCoV-2 viral load among COVID-19 patients.
METHODSA systematic review of effectiveness was conducted. The databases included the Cumulative Index to Nursing and Allied Health Literature (CINAHL), MEDLINE, Google Scholar, Embase, The Cochrane Library, and BMJ Clinical Evidence and recruitment studies published between 2021-2023. The Joanna Briggs Institute (JBI) Critical Appraisal Checklist for Randomized Controlled Trials was used to evaluate the risk of bias.
RESULTSA total of 523 patients from 10 clinical studies were included in this analysis. The review demonstrated the potential effectiveness of CHX Mouthwash in reducing SARS-CoV-2 viral load among COVID-19 patients admitted to a healthcare facility.
CONCLUSION AND RECOMMENDATIONSThis review summarized current evidence on the antimicrobial and virucidal effects of chlorhexidine (CHX) in clinical settings. Findings indicated that CHX mouthwash may help reduce the SARS-CoV-2 viral load among hospitalized COVID-19 patients. However, variations in study design and outcomes, along with the narrative synthesis approach, limited causal interpretation. The results may guide clinical nurses and healthcare institutions in updating infection control practices and oral care protocols for patients with COVID-19.
Human ; Chlorhexidine ; Covid-19 ; Sars-cov-2 ; Health
10.Prokaryotic expression, purification and immunogenicity of SARS-CoV-2 omicron variant nucleocapsid protein.
Zewen TU ; Quansheng WANG ; Shiguo LIU ; Haosen LIU ; Chunyan ZENG ; Juanjuan XIE ; Mingzhi LI ; Jingcai LI ; Min WANG ; Shiqi WENG ; Lumei KANG ; Lingbao KONG
Chinese Journal of Cellular and Molecular Immunology 2025;41(8):735-743
Objective The study aims to investigate the immunological functions of the nucleocapsid (N) protein of the novel coronavirus Omicron (BA.1, BA.2) and evaluate the differences among different N proteins of mutant strains in immunogenicity. Methods By aligning sequences, the mutation sites of the Omicron (BA.1, BA.2) N protein relative to prototype strain of the novel coronavirus (Wuhan-Hu-1) were determined. The pET-28a-N-Wuhan-Hu-1 plasmid was used as template to construct pET-28a-BA.1/BA.2-N through single point mutation or homologous recombination. The three kinds of N protein were expressed in prokaryotic system, purified through Ni-NTA affinity chromatography, and then immunized into mice. The titer and reactivity of the polyclonal antibody, as well as the expression level of IL-1β and IFN-γ in mouse spleen cells, were detected using indirect ELISA and Western blot assay. Results The constructed prokaryotic expression plasmids were successfully used to express the Wuhan-Hu-1 N, BA.1 N, and BA.2 N proteins in E.coli BL21(DE3) at 37 DegreesCelsius for 4 hours. The indirect ELISA test showed that the titers of polyclonal antibody prepared by three N proteins were all 1:51 200. All three N proteins can increase the expression of IFN-γ and IL-1β cytokines, but the effect of Omicron N protein in activing two cytokines was more obvious than that of Wuhan-Hu-1 N protein. Conclusion The study obtained three new coronavirus N proteins and polyclonal antibodies, and confirmed that mutations in the amino acid sites of the N protein can affect its immunogenicity. This provides a basis for developing rapid diagnostic methods targeting N protein of different novel coronavirus variants.
Animals
;
Mice
;
SARS-CoV-2/genetics*
;
Coronavirus Nucleocapsid Proteins/immunology*
;
Nucleocapsid Proteins/isolation & purification*
;
COVID-19/immunology*
;
Antibodies, Viral/immunology*
;
Mice, Inbred BALB C
;
Interferon-gamma/metabolism*
;
Interleukin-1beta/metabolism*
;
Female
;
Escherichia coli/metabolism*
;
Mutation
;
Humans


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