Genomic analysis of three SARS-CoV-2 waves in west Sumatra: Evolutionary dynamics and variant clustering.

Background: The Severe Acute Respiratory Syndrome Coronavirus 2 virus (SARS-CoV-2) has been undergoing evolutionary changes to improve its ability to thrive within human hosts, leading to the emergence of specific variants associated with subsequent waves of the coronavirus diseases 2019 (COVID-19) pandemic. Indonesia has grappled with the effects of this pandemic and subsequent waves affecting various regions, including West Sumatra. Although located outside Java island epicenter, West Sumatra experienced significant COVID-19 transmission, especially during the third wave in early 2022. Objective: This study aimed to investigate the genetic evolution and epidemiological dynamics of SARS-CoV-2 variants in West Sumatra throughout the three pandemic waves. Methods: We conducted a genotyping study retrospectively using 278 COVID-19 patient samples from 2020 to 2022. The Real-Time Quantitative Reverse Transcription PCR (RT-qPCR) was used for screening, and whole-genome sequence analysis was conducted through the Illumina MiSeq instrument. Result: The analysis revealed distinct patterns in the prevalence of viral lineages across the waves. The initial wave was predominated by clade 20A (77,4 %) especially lineage B.1.466.2 (50 %). The second wave was marked by a significant emergence of the Delta variant (72,5 %), particularly lineage AY.23 (81,1 %), originating from India, with subsequent local evolution leading to the formation of distinct clusters. We found that about 96,7 % of the third wave variant was dominated by Omicron variants, especially the generation of lineages BA.1 and BA.2, demonstrating widespread global dissemination and local variant development. Phylogenetic analysis indicated a close relatedness of West Sumatra variants to those from Malaysia and other parts of Indonesia, highlighting regional transmission dynamics and potential sources of variant introductions. Conclusion: This study has identified unique variant clusters within each wave, suggesting distinct evolutionary pathways and local adaptations. These findings provide valuable insights into the genomic landscape of SARS-CoV-2 in West Sumatra and emphasize the crucial role of ongoing genomic surveillance in tracking viral changes and guiding public health measures.

Dynamic Evolution of SARS-CoV-2 in West Sumatra: Analyzing S Gene Mutations Across Variants and Their Impact on Public Health and Vaccine Strategies.

<b>Background and Objective:</b> The global SARS-CoV-2 pandemic highlights the importance of tracking virus evolution through genomic surveillance, especially concerning mutations in the SARS-CoV-2 spike protein, crucial for vaccine development. Despite global concern over variants, regions like West Sumatra, Indonesia, lack thorough genomic analysis, prompting this study to analyze S gene mutations across three pandemic waves in West Sumatra. <b>Materials and Methods:</b> Next-generation sequencing was conducted through the Illumina MiSeq instrument to leverage a dataset of 352 anonymized samples collected between March, 2020 and November, 2022 and rigorous analysis of S gene mutation using CLC Genomics Workbench^® 21 version 21.0.3 were employed. Statistical analyses assessed mutation prevalence over time, exploring associations with clinical outcomes. <b>Results:</b> The findings revealed significant variability in mutation profiles across different variants. Notably, the Omicron variant (21K) exhibited a high mutation rate, suggesting enhanced immune evasion capabilities. Comparative analysis highlighted evolutionary trends, from early variants with fewer mutations to highly adapted forms like Delta (21I) and Omicron. The dynamic nature of SARS-CoV-2 evolution underscores the importance of continuous surveillance, rapid public health response and vaccine adaptation. <b>Conclusion:</b> This study contributes valuable insights into the virus's evolving landscape, emphasizing the need for ongoing research, global collaboration and adaptable vaccine strategies to manage the evolving threat of COVID-19 effectively.

The exploration of glucocorticoid pathway based on disease severity in COVID-19 patients.

Systemic inflammation is a hallmark of Coronavirus Disease 2019 (COVID-19) and is the key to the pathophysiology of its severe cases with host cytokine involvement. Glucocorticoids can moderate this inflammatory effect due to receptor binding (NRC31-the gene encoded), influencing the expression of effector genes and pro-inflammatory cytokines. Another important pathway in the processes of the immune and inflammatory responses is nuclear factor-κB (NF-κB) signaling (NFKBIA-the gene encoded). We aimed to explore the expression of genes in the glucocorticoid pathway in mild and severe COVID-19. We performed a cross-sectional, observational study on COVID-19 cases, assessing the expression of RNA in white blood cells. The Illumina® platform was used for RNA sequencing, and FASTQ data were quality-checked with Multiqc. The raw data were analyzed using CLC Genomics Workbench®. Our study included 23 patients with severe COVID-19 and 21 patients with mild COVID-19 with an average age of 49.9 ± 18.2 years old. The NR3C1 and NFKBIA expressions did not show a significantly significant difference between groups (log2 fold change 0.5, p = 0.1; 0.82, p = 0.09). However, the expressions of TSC22D3, DUSP-1, JAK-1 and MAPK-1 were significantly higher in mild cases (log2 fold change 1.3, p < 0.001; 2.6, p < 0.001; 0.9, p < 0.001; 1.48, p-value<0.001; respectively). Furthermore, the TNF, IL-1ß, and IL-6 expressions were significantly lower in mild cases (log2 fold change 4.05, p < 0.001; 3.33, p < 0.001; 6.86, p < 0.001; respectively). In conclusion, our results showed that although the NRC31 and NFKBIA expressions did not show a statistically significant difference between groups, the expression of TSC22D3 was higher in mild cases. These results highlight the importance of effector genes, specifically TSC22D3, in combatting systemic inflammation. Our recent findings have the potential to lead to the identification of novel pharmacological targets that could prove to be vital in the fight against diseases associated with inflammation.

Effects of Lactococcus lactis on colorectal cancer in various terms: a narrative review.

Introduction: Colorectal cancer is one of the most common cancers with an increasing number of cases. Various studies have found an association between the gut microbiota balance and colorectal cancer incidence. Lactococcus lactis is a probiotic bacterium found in fermented foods, particularly yogurt and cheese. This probiotic has been shown to reduce various anti-inflammatory and pro-inflammatory agents that trigger cancer, such as interleukin (IL)-6, IL-18, tumour necrosis factor-alpha (TNF-α), and natural killer (NK) cells. Methods: Full-text articles and original research published in the last ten years were used as references, and "Lactococcus and colorectal cancer" as keywords. The reference search is on several databases, such as PubMed, ScienceDirect, ProQuest, and Nature. Searching results obtained eleven articles. Discussion: Lactococcus lactis does have a perfect role in suppressing cancer cells. Lactococcus lactis has anti-proliferative effects associated with decreased cyclin D1 expression in SW480 cell lines, decreased NK cells, reduced cancer cell viability, decreased IL-8 levels, and decreased IL-6. Conclusion: Lactococcus lactis contains nisin, which can suppress various gene, protein, and cytokine expressions that play a role in cancer cell growth. Probiotics can inhibit colorectal cancer without significant side effects.

Investigation of Initial Viral Loads and Patient Characteristics as Predictors of COVID-19 Outcomes: A Retrospective Cohort Study.

Limited evidence exists on whether initial viral load and patient characteristics can predict unfavorable outcomes in future outbreaks of coronavirus disease 2019 (COVID-19). This retrospective cohort study examined the relationship between the initial viral load, patient characteristics, and outcomes during the second-wave COVID-19 outbreak in West Sumatra, Indonesia. We analyzed the COVID-19 patients admitted to a secondary hospital between the 1 June 2021 and the 31 August 2021. The initial viral load was determined using the real-time quantitative-polymerase chain reaction (RT-qPCR) cycle threshold (Ct) value, categorized as low (LIVL, Ct > 20) or high (HIVL, Ct ≤ 20). Multivariate logistic regression was used to assess the relationship between the initial viral load, age, sex, vaccination status, comorbidities, and outcomes, including disease severity, hospital stay length, ICU admission, invasive ventilation, and in-hospital mortality. Receiver operating characteristic (ROC) curves and the area under the curve (AUC) were used to assess the diagnostic performance of the initial Ct values in predicting COVID-19 outcomes. The study included 373 patients (median age [range]: 48 [0-94]; male: 40.21%; HIVL: 34.85%; unvaccinated: 86.06%; comorbidities: 52.01%). The HIVL patients significantly had a lower risk of developing severe/critical outcomes (OR: 0.506; 95% CI: 0.310-0.825; p = 0.006) and needing invasive ventilation (OR: 0.290; CI: 0.098-0.854; p = 0.025). The Ct value used to indicate severe/critical outcomes was 23.57. More severe outcomes were significantly observed in LIVL patients, those aged >60 years, males, unvaccinated individuals, and those with comorbidities. This study emphasizes the importance of primary prevention, early screening, and immediate care for COVID-19 in saving lives.

The role of the glucocorticoid receptor and its impact on steroid response in moderate-severe COVID-19 patients.

The effect of corticosteroid therapy in COVID-19 patients is mediated by its suppressive effect on the regulations of inflammatory response. However, its clinical outcome is often unpredictable. This study aimed to explore the role of glucocorticoid receptors in corticosteroid response in Moderate-Severe COVID-19 patients. In this cross-sectional study, we attempted to find the relationship between the expression of the glucocorticoid receptor (encoded by NR3C1), the variation of glucocorticoid receptors isoform, and the mutations of glucocorticoid receptors exon with clinical response to corticosteroids. In addition, the relationship between glucocorticoid receptors expression and the expression of IκBα (encoded by NFKBIA) and glucocorticoid-induced leucine zipper protein (GILZ; encoded by TSC22D3) as steroid pathways was also evaluated. Thirty-four COVID-19 patients were studied. Blood was drawn before and on day 5 of corticosteroid treatment. Glucocorticoid receptors expression, isoform, and mutation were determined by RNA sequencing from white blood cells. Based on the improvement of clinical and oxygen status, patients were classified into responder and non-responder groups. Of thirty-four patients, 23 (67.6%) showed excellent responses to corticosteroids, and 11 (32.4%) were non-responders. The NR3C1 gene expression was significantly higher in the responsive group at baseline and after five days of glucocorticoid treatment. Isoform variant and mutation of glucocorticoid receptors did not correlate with clinical response. The expression of IκBα and GILZ correlated positively with glucocorticoid receptors expression. This study elucidates the relationship between glucocorticoid receptor expression with therapeutic responses to corticosteroids in moderate-severe COVID-19.

Human Papillomavirus Type 16 L2 Gene Sequence Variation Analysis in Indonesian Cervical Cancer Specimens.

BACKGROUND: Human papillomavirus type 16 (HPV16) is the most prevalent etiology of cervical cancer in Indonesian women. The L2 minor capsid protein has considerable potential as a broad-protective antigen target of the cervical cancer vaccine strategies, yet the data on L2 gene variation is still minimal. In this research, we determined the variations of the HPV16 L2 gene sequences in Indonesian cervical cancer specimens. METHODS: We cross-sectionally observed 23 DNA isolates of HPV16 positive cervical cancer specimens stored in the laboratory of the Center for Diagnostic and Research on Infectious Diseases (PDRPI Lab), Faculty of Medicine, Universitas Andalas, Padang, Indonesia. We detected and amplified the HPV16 L2 gene sequences in the samples, followed by sequencing, DNA alignment, single nucleotide polymorphisms (SNPs) analysis, and phylogenetic tree reconstruction. RESULTS: As many as 35 SNPs were found, consist of 18 synonymous SNPs (sSNPs) and 17 non-synonymous SNPs (nsSNPs). Amino acid variations were mostly detected at S269P (100%) and L330F (43.48%) with no variation in the immuno-protective region near L2 N-terminus. A total of 5 HPV16 phylogenetic sub-lineages were found closely related to A1 (n=5), A2 (n=12), A3 (n=2), A4 (n=3), and C (n=1). CONCLUSION: The variations of HPV16 L2 gene sequences are mainly located in the central region of the L2 sequences, and the cross-protective region near the L2 N-terminus is remarkably conserved. This study should enhance the information about HPV16 L2 gene variation in Indonesia.

Prevalence Oncogenic Human Papillomavirus in Cervical Cancer Patients in Riau Province Indonesia.

Background: Cervical cancer is the fourth most deadly cancer in the world, and it is caused by infection of high-risk subtypes of Human PapillomaVirus (HPV) in most cases. The aims of this study were to determine the prevalence oncogenic HPV in cervical cancer patients in Riau Province Indonesia and to determine the clinical manifestation of HPV in cervical cancer patients in Riau Province Indonesia. Methods: This research was a descriptive study conducted at Arifin Achmad General Hospital Riau from February to August 2018 which aimed to analyze HPV genotype prevalence oncogenic of cervical cancer patients. Results: This study showed out 86 of 110 women (78.1%) were found HPV positive, and the most common genotype of HPV was HPV 16 (38.2%). The average age of cervical cancer patient was 50 years old, and the average number of parities was 4 times. The majority of participants were married at the age before 20 years (77.3%) and had low educational background (64.5%). Vaginal bleeding happened in more than half of the participant as major clinical manifestation (72.7%), followed by fluor albus (72.7%), pelvic pain (60.2%) and fatigue (65.9%). Conclusion: The most common HPV genotype in Riau Province was HPV type 16 and the most common clinical symptoms of cervical cancer patient were vaginal bleeding, fluor albus, pelvic pain and fatigue.

Detecting Human Papillomavirus Type 16 in Cervical Cancer Patients with Molecular Variation of Gene L1 in Riau Province Indonesia.

BACKGROUND: Cervical cancer is the second most deadly cancer in the world after breast cancer. The cancer is caused by infection of high risk Human Papillomavirus (HPV) type 16. It is often found in cervical cancer of which the genome structure is composed of L1 proteins. The L1 protein makes up the viral capsid that has an important role in causing the cervical epithelium. Several studies have found the differences in HPV nucleotides variants that lead to changes in amino acids that disrupt the structure, the natural function of the virus itself, and ultimately lead to changes in biological functions including host immunological recognition. Variation of the L1 gene also affects the effectiveness of existing vaccines. METHODS: This research was a descriptive study conducted at the laboratory of microbiology, the Faculty of Medicine, Universitas Riau, Pekanbaru from February to August 2018. The study was aimed at looking at the molecular variations of the L1 HPV type 16 gene and examining phylogenic kinship. RESULTS: The SNPs (Single Nucleotide Polymorphism) which occurred in 26 sample isolates are the substitution of C/G (6240), A/G (6432), T/G (6686), C/T (6824). These variations also cause changes in amino acids, insertion of ATC nucleotide bases (6902), and deletions of GAT bases (6954). CONCLUSION: There are  molecular variations of the L1 HPV type 16 gene which can cause different host immune responses. Phylogenic kinship of HPV type 16 isolate in Riau is similar to  Asian-American isolate.

Epidermal Growth Factor and Adenosine Triphosphate Induce Natrium Iodide Symporter Expression in Breast Cancer Cell Lines.

AIM: This study aims to investigate the effect of ATP, EGF and combination of those two to the Natrium Iodide Symporter (NIS) expression in MCF7, SKBR3 and HaCaT cell lines. METHODS: MCF7, SKBR3 and HaCaT cell lines were treated with ATP, EGF and combination of those two for 6, 12 and 24 hours. The expression of NIS mRNA was measured through quantitative-reverse transcription-polymerase chain reaction (qRT-PCR). The NIS protein expression was confirmed by immunocytofluorescence. RESULTS: NIS mRNA was expressed in SKBR3 and HaCaT cell lines but not in MCF7. The levels of NIS mRNA expression, after treatment by epidermal growth factor (EGF), adenosine Tri-Phosphate (ATP) or the combination of both for 6 and 12 hours were not significantly different from those of untreated cells. However, the treatment by a combination of ATP and EGF for 24 hours increases the level of NIS mRNA expression by 1.6 fold higher than that of the untreated cells (1.6241 ± 0.3, p < 0.05) and protein NIS expression increase significantly by the treatment than untreated cells (P < 0.05). CONCLUSION: The level of NIS expression varies among the different subtypes of breast cancer cell lines. MCF7 cell line is representing the luminal A subtype of breast cancer does not express NIS. Only SKBR3 cell line express NIS and this subtype might be suitable to receive radioiodine therapy as those cells expressing NIS. A combination treatment of EGF and ATP increases the expression of NIS mRNA and protein at the membrane in SKBR3 cells.