Cytokine profiles in dengue fever and dengue hemorrhagic fever: A study from Indonesia.

Recent studies have demonstrated that cytokine dysregulation has a critical role in the pathogenesis of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). The aim of this study was to investigate the association between tumor necrosis factor (TNF- α), interleukin 6 (IL-6), interleukin 10 (IL-10), and interleukin 17 (IL-17) with infection status, and severity of dengue. A prospective cross-sectional study was conducted at three hospitals in Gianyar regency and Denpasar municipality, Bali, Indonesia, from June to December 2022. Sixty-four dengue infected patients were involved. Patients' serum was tested for dengue infection using NS1 antigen rapid test, dengue virus immunoglobulin M (IgM) and immunoglobulin G (IgG) test, and reverse transcription polymerase chain reaction (RT-PCR). Cytokine levels (TNF-α, IL-6, IL-10, and IL-17) were measured using enzyme-linked immunosorbent assay (ELISA). Infection status was determined by combining serological and RT-PCR results, categorizing patients into primary and secondary infections. The present study found that DF patients had lower TNF-α, IL-6, and IL-17 but higher IL-10 levels compared to DHF patients (p<0.001). Elevated TNF-α, IL-6, and IL-17 levels were higher in secondary infection, while IL-10 level was higher in primary infection (p<0.001). In conclusion, cytokines play a crucial role in the interplay between cytokine dysregulation and dengue infection dynamics.

Relationship between neutrophil-lymphocyte ratio and platelet-lymphocyte ratio with the severity of COVID-19.

Coronavirus disease 2019 (COVID-19) is an infectious disease that spreads rapidly causing a high case fatality rate in vulnerable populations. Neutrophil-lymphocyte ratio (NLR) and platelet-lymphocyte ratio (PLR) are known to be inflammatory biomarkers in certain infections. The aim of this study was to determine the relationship between NLR and PLR with the severity of COVID-19. A cross-sectional study was conducted at Tabanan Regency General Hospital, Bali, Indonesia, from January 2021 to December 2022. All patients included in the study tested positive for COVID-19 by real-time polymerase chain reaction (RT-PCR), aged 18-50 years with no comorbid. Laboratory examinations were carried out on admission. The patients were categorized into two groups based on the severity: moderate and severe/critical. The Mann-Whitney test was used to determine the association between NLR and PLR with the severity of COVID-19. A total of 104 patients were included in the study, the majority of COVID-19 patients had moderate (77.9%) severity. The average NLR was 5.8 and the PLR was 21.7. There was a significant relationship between NLR (p=0.002) and PLR (p=0.001) with the severity of COVID-19. The defined cut-off values of NLR and PLR were ≥3.8 and ≥106, yielding sensitivities of 95% and 70%, and specificities of 74% and 50%, respectively. This study highlights the promising role of NLR and PLR as predictive biomarkers to assess COVID-19 severity.

Anti-inflammatory activities of flavonoid derivates.

Background and purpose: Flavonoids are a group of phytochemicals found abundantly in various plants. Scientific evidence has revealed that flavonoids display potential biological activities, including their ability to alleviate inflammation. This activity is closely related to their action in blocking the inflammatory cascade and inhibiting the production of pro-inflammatory factors. However, as flavonoids typically have poor bioavailability and pharmacokinetic profile, it is quite challenging to establish these compounds as a drug. Nevertheless, progressive advancements in drug delivery systems, particularly in nanotechnology, have shown promising approaches to overcome such challenges. Review approach: This narrative review provides an overview of scientific knowledge about the mechanism of action of flavonoids in the mitigation of inflammatory reaction prior to delivering a comprehensive discussion about the opportunity of the nanotechnology-based delivery system in the preparation of the flavonoid-based drug. Key results: Various studies conducted in silico, in vitro, in vivo, and clinical trials have deciphered that the anti-inflammatory activities of flavonoids are closely linked to their ability to modulate various biochemical mediators, enzymes, and signalling pathways involved in the inflammatory processes. This compound could be encapsulated in nanotechnology platforms to increase the solubility, bioavailability, and pharmacological activity of flavonoids as well as reduce the toxic effects of these compounds. Conclusion: In Summary, we conclude that flavonoids and their derivates have given promising results in their development as new anti-inflammatory drug candidates, especially if they formulate in nanoparticles.

Dengue seroprevalence study in Bali.

INTRODUCTION: Dengue infection poses significant public health problems in tropical and subtropical regions worldwide. The clinical manifestations of dengue vary from asymptomatic to severe dengue manifestations. This serological survey highlighted the high incidence of asymptomatic cases. This study aimed to determine the prevalence of dengue in healthy and ill adults in Bali. METHODS: Cross-sectional seroprevalence surveys were performed between July 2020 and June 2021 among healthy and ill adults in Denpasar Bali. Blood samples were collected from 539 randomly selected urban sites in Denpasar. Immunoglobulin G antibodies against the dengue virus were detected in serum using a commercial enzyme-linked immunosorbent assay kit. RESULTS: Overall, the dengue seroprevalence rate among the 539 clinically healthy and ill adults was high (85.5%). The median age was 34.1 (18-86.1). Most of the participants in the study were younger than 40 years (61.2%). Men were the dominant sex (54.5%). The study found a significant association between dengue seropositivity among people aged > 40 years and healthy status (p = 0.005; odds ratio [OR] = 0.459 and p < 0.001; OR = 0.336, respectively). The study reported that as many as 60% of the subjects had a history of previously suspected dengue infection. This study reflected the proportion of asymptomatic dengue patients requiring better assessment with a serological test. CONCLUSION: The current study highlighted that real cases of dengue infection may be higher than reported, with a high prevalence of dengue seropositivity and a relatively dominant proportion of asymptomatic cases. The study guides physicians to be aware of every dengue infection in tropical countries and prevent the spread of the disease.

The role of CD4+, CD8+, CD4+/CD8+ and neutrophile to lymphocyte ratio in predicting and determining COVID-19 severity in Indonesian patients.

BACKGROUND: Biomarkers that are cost-effective and accurate for predicting severe coronavirus disease 2019 (COVID-19) are urgently needed. We would like to assess the role of various inflammatory biomarkers on admission as disease severity predictors and determine the optimal cut-off of the neutrophile-to-lymphocyte ratio (NLR) for predicting severe COVID-19. METHODS: We conducted a cross-sectional study in six hospitals in Bali and recruited real-time PCR-confirmed COVID-19 patients aged >18 y from June to August 2020. Data collection included each patient's demographic, clinical, disease severity and hematological data. Multivariate and receiver operating characteristic curve analyses were performed. RESULTS: A total of 95 Indonesian COVID-19 patients were included. The highest NLR among severe patients was 11.5±6.2, followed by the non-severe group at 3.3±2.8. The lowest NLR was found in the asymptomatic group (1.9±1.1). The CD4+ and CD8+ values were lowest in the critical and severe disease groups. The area under the curve of NLR was 0.959. Therefore, the optimal NLR cut-off value for predicting severe COVID-19 was ≥3.55, with sensitivity at 90.9% and a specificity of 16.7%. CONCLUSIONS: Lower CD4+ and CD8+ and higher NLR values on admission are reliable predictors of severe COVID-19 among Indonesian people. NLR cut-off ≥3.55 is the optimal value for predicting severe COVID-19.

The role of mediator suppressor of cytokine signaling (SOCS), toll-like receptor 3 (TLR-3) and nuclear factor kappa B (NFκB) on cytokine production during dengue virus infection.

Inability to understand the pathogenesis of severe dengue, in particular the control mechanism of immune responses, has led to high mortality rate for patients with dengue shock syndrome (DSS). The aim of this study was to determine the control mechanism of cytokine production by mediator suppressor of cytokine signaling (SOCS), toll-like receptor 3 (TLR-3) and nuclear factor kappa B (NFκB) during DENV infection. Peripheral blood mononuclear blood cells (PBMC), isolated from healthy individuals, were infected with dengue virus (DENV)-2 strain SJN-006 Cosmopolitan genotype (isolated from Bali, Indonesia). The relative gene expression of SOCS-3, TLR-3, NFκB, and the cytokine genes (interleukin (IL)-6, IL-8, interferon inducible protein 10 (IP-10), and macrophage inflammatory protein-1 beta (MIP-1ß)) were measured using qRT-PCR at 6, 12 and 24 hours post infection (hpi). Student t-test and Mann-Whitney test were used to compare the gene expressions while causal correlations were analyzed using regression test and path analyses. DENV-2 infection increased the gene expression of SOCS-3, TLR-3, and NFκB after 12 and 24 hpi. The expression of IL-6, IL-8, IP-10, and MIP-1ß genes was increased and peaked at different times post-infection. NFκB and SOCS-3 genes likely have role in the upregulation of IL-8 and IL-6 gene expression, respectively. MIP-1ß gene expression was significantly induced by both NFκB and SOCS-3. In conclusion, our study suggested that SOCS-3, TLR-3, and NFκB are important in regulating the production of IL-6, IL-8, IP-10, MIP-1ß during early phase of DENV-2 infection. This enriches our understanding on pathogenesis pathway of DENV-associated cytokine storm.

Clinical characteristics and outcomes of COVID-19 patients in Bali, Indonesia.

INTRODUCTION: The spectrum of illness and outcomes of coronavirus disease 2019 (COVID-19) patients may vary. This study reports the characteristics of COVID-19 patients in Bali, Indonesia, and evaluates the diagnostic value of their clinical symptoms. METHOD: This observational study was conducted in eight hospitals. The patients were classified as non-severe COVID-19, severe COVID-19, and non-COVID-19. Demographics, clinical, laboratory, and radiologic characteristics, and outcomes of COVID-19 patients were collected. Factors associated with the severity and outcomes were assessed using the chi-squared test or ANOVA when appropriate. We also compared the clinical features of non-severe COVID-19 and non-COVID-19 patients to evaluate the diagnostic accuracy. RESULTS: This study included 92 patients: 41 non-COVID-19 and 51 COVID-19 patients, comprising 45 non-severe and six severe cases. The most common symptoms of COVID-19 were cough (47.1%), fever (31.0%), and dyspnea (25.3%). Cough, fatigue, and anosmia have high accuracy, and combining these complaints in clinical diagnostics offered a higher accuracy in predicting COVID-19 patients (60.1%). We found lower lymphocyte counts and interleukin-1R levels and higher levels of C-reactive protein, interleukin-6, and interleukin-8 in severe compared than in non-severe COVID-19 patients. Lactate dehydrogenase was associated with intensive care unit admission and ventilator use, while other markers such as neutrophil-lymphocyte ratio, C-reactive protein, and interleukin-6 were not. CONCLUSION: A battery of symptoms, including cough, fatigue, and anosmia, is likely associated with COVID-19 in Bali. Clinicians should be aware of these symptoms to ensure a prompt diagnostic test for COVID-19, beyond other causes of acute febrile illnesses.

Combining rapid diagnostic tests to estimate primary and post-primary dengue immune status at the point of care.

BACKGROUND: Characterising dengue virus (DENV) infection history at the point of care is challenging as it relies on intensive laboratory techniques. We investigated how combining different rapid diagnostic tests (RDTs) can be used to accurately determine the primary and post-primary DENV immune status of reporting patients during diagnosis. METHODS AND FINDINGS: Serum from cross-sectional surveys of acute suspected dengue patients in Indonesia (N:200) and Vietnam (N: 1,217) were assayed using dengue laboratory assays and RDTs. Using logistic regression modelling, we determined the probability of being DENV NS1, IgM and IgG RDT positive according to corresponding laboratory viremia, IgM and IgG ELISA metrics. Laboratory test thresholds for RDT positivity/negativity were calculated using Youden's J index and were utilized to estimate the RDT outcomes in patients from the Philippines, where only data for viremia, IgM and IgG were available (N:28,326). Lastly, the probabilities of being primary or post-primary according to every outcome using all RDTs, by day of fever, were calculated. Combining NS1, IgM and IgG RDTs captured 94.6% (52/55) and 95.4% (104/109) of laboratory-confirmed primary and post-primary DENV cases, respectively, during the first 5 days of fever. Laboratory test predicted, and actual, RDT outcomes had high agreement (79.5% (159/200)). Among patients from the Philippines, different combinations of estimated RDT outcomes were indicative of post-primary and primary immune status. Overall, IgG RDT positive results were confirmatory of post-primary infections. In contrast, IgG RDT negative results were suggestive of both primary and post-primary infections on days 1-2 of fever, yet were confirmatory of primary infections on days 3-5 of fever. CONCLUSION: We demonstrate how the primary and post-primary DENV immune status of reporting patients can be estimated at the point of care by combining NS1, IgM and IgG RDTs and considering the days since symptoms onset. This framework has the potential to strengthen surveillance operations and dengue prognosis, particularly in low resource settings.

Molnupiravir: A lethal mutagenic drug against rapidly mutating severe acute respiratory syndrome coronavirus 2-A narrative review.

Broad-spectrum antiviral agents targeting viral RNA-dependent RNA polymerase (RdRp) are expected to be a key therapeutic strategy in the ongoing coronavirus disease 2019 (COVID-19) pandemic and its future variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes COVID-19. Molnupiravir is a nucleoside analog that in vivo experiments have been reported to inhibit the replication of SARS-CoV-2, the virus that causes COVID-19. Clinical trials of molnupiravir as a therapy for patients with mild-to-moderate COVID-19 also suggest its significant therapeutic efficacy in comparison to placebo. Molnupiravir is lethally mutagenic against viral RNA, but its effect on host cell DNA is being questioned. Herein, the safety concerns of molnupiravir are discussed with recent findings from published reports and clinical trials. The unchanged efficacy of molnupiravir against mutated SARS-CoV-2 variants is also highlighted. With its administration via the oral route, molnupiravir is expected to turn the tide of the COVID-19 pandemic.

Titer disparity of anti-Spike receptor binding domain SARS-CoV-2 antibody between vaccinated and naturally infected individuals.

In conjunction with other health promotion strategies, vaccination of coronavirus disease 2019 (COVID-19) is a strategy to alleviate the burden of infection. The aim of this study was to determine the differences in antibody response strength between individuals who received COVID-19 vaccination and those who had a natural infection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A cross-sectional study was conducted among post-natural confirmed COVID-19 infection and immunized people in Bali, Indonesia. The vaccination was using Sinovac-CoronaVac with two-weeks interval between the two vaccine doses. To measure the level of anti-Spike receptor binding domain (SRBD) of SARS-CoV-2 antibody, we used Roche electro- chemiluminescence immunoassay (ECLIA) platform. Blood samples were obtained before and 28 days after first immunization in the vaccinated group, as well as two weeks after hospital discharge in the confirmed COVID-19 patients based on real-time reverse transcription polymerase chain reaction (RT-PCR). A total of 58 confirmed COVID-19 patients and 60 vaccinated individuals were included. On the 28th day after the initial vaccination, the seroconversion rate among vaccinated individuals was 91.67%. The mean titer of anti-SRBD SARS-CoV-2 antibody among vaccinated participants was 63.62±82.57 IU/mL (ranged between 0 IU/mL and 250 IU/mL). The mean titer among naturally infected group was 188.47±94.57 IU/mL (ranged between 4.25 IU/mL to 250 IU/mL) regardless the severity of COVID-19. Our data suggested that the titer of anti- SRBD SARS-CoV-2 antibody was significantly higher in naturally infected individuals compared to those who received COVID-19 vaccination (p<0.001). These data suggest that not all individuals vaccinated with Sinovac COVID-19 had protective level of anti- SRBD SARS-CoV-2 antibody and booster dose of heterologous vaccine maybe required.

Global prevalence of prolonged gastrointestinal symptoms in COVID-19 survivors and potential pathogenesis: A systematic review and meta-analysis.

Background: This study aimed to determine the cumulative prevalence of prolonged gastrointestinal (GI) symptoms, including nausea, vomiting, diarrhea, lack of appetite, abdominal pain, and dysgeusia, in survivors of both mild and severe COVID-19 worldwide and to discuss the potential pathogenesis.   Methods: Three databases (PubMed, Scopus, and Web of Science) were searched for relevant articles up to January 30, 2021. Data on study characteristics, clinical characteristics during follow-up, the number of patients with prolonged GI symptoms, and total number of COVID-19 survivors were retrieved according to PRISMA guidelines. The quality of eligible studies was assessed using the Newcastle-Ottawa scale. The pooled prevalence of specific prolonged GI symptoms was calculated and the association between COVID-19 severity and the occurrence of prolonged GI symptoms was assessed if appropriate.   Results: The global prevalence of prolonged nausea was 3.23% (95% CI: 0.54%-16.53%) among 527 COVID-19 survivors. Vomiting persisted in 93 of 2,238 COVID-19 survivors (3.19%, 95% CI: 1.62%-6.17%) and prolonged diarrhea was found in 34 of 1,073 survivors (4.12%, 95% CI: 1.07%-14.64%). A total of 156 patients among 2,238 COVID-19 survivors (4.41%, 95% CI: 1.91%-9.94%) complained of persistent decreased or loss of appetite. The cumulative prevalence of prolonged abdominal pain was 1.68% (95% CI: 0.84%-3.32%), whereas persistent dysgeusia was identified in 130 cases among 1,887 COVID-19 survivors (7.04%, 95% CI: 5.96%-8.30%). Data was insufficient to assess the relationship between COVID-19 severity and the occurrence of all prolonged GI symptoms.   Conclusion: Persistent GI symptoms among COVID-19 survivors after discharge or recovery raises a concern regarding the long-term impact of the COVID-19 infection on the quality of life of the survivors. Despite several potential explanations proposed, studies that aim to follow patients after recovery from COVID-19 and determine the pathogenesis of the prolonged symptoms of COVID-19 survivors are warranted.   PROSPERO registration: CRD42021239187.

Pulmonary arterial hypertension post COVID-19: A sequala of SARS-CoV-2 infection?

It has been suggested that pulmonary arterial hypertension (PAH) could be a potential sequela of coronavirus disease 2019 (COVID-19) in particular in those with hypertension; however, development of PAH after the course of COVID-19 in normotensive individuals are rarely reported. Here, we report a patient who developed PAH two months post-COVID-19. The patient was a 55-year-old female and normotensive, tested positive for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), developed mild respiratory distress syndrome and necessitated continuous positive airway pressure during the treatment in the hospital. After two months discharged from the hospital with RT-PCR negative for SARS-CoV-2, the patient presented with exertional dyspnea, dry cough, fatigue and episodes of syncope during exertion. Based on clinical presentation, electrocardiography, computed tomography, and transthoracic echocardiography assessment, PAH diagnosis was made. To our knowledge, this is a rare PAH case and this highlights the possible of PAH as sequala that might present in post COVID-19 patients.

Assessment of dengue and COVID-19 antibody rapid diagnostic tests cross-reactivity in Indonesia.

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic remains ongoing around the world, including in areas where dengue is endemic. Dengue and COVID-19, to some extent, have similar clinical and laboratory features, which can lead to misdiagnosis, delayed treatment and patient's isolation. The use of rapid diagnostic tests (RDT) is easy and convenient for fast diagnosis, however there may be issues with cross-reactivity with antibodies for other pathogens. METHODS: We assessed the possibility of cross-reactivity between SARS-CoV-2 and dengue antibodies by: (1) testing five brands of COVID-19 IgG / IgM RDTs on 60 RT-PCR-confirmed dengue samples; (2) testing 95 RT-PCR-confirmed COVID-19 samples on dengue RDT; and (3) testing samples positive for COVID-19 IgG and/or IgM on dengue RDT. RESULTS: We observed a high specificity across all five brands of COVID-19 RDTs, ranging from 98.3 to 100%. Out of the confirmed COVID-19 samples, one patient tested positive for dengue IgM only, another tested positive for dengue IgG only. One patient tested positive for dengue IgG, IgM, and NS1, suggesting a co-infection. In COVID-19 IgG and/or IgM samples, 6.3% of COVID-19 IgG-positive samples also tested positive for dengue IgG, while 21.1% of COVID-19 IgM-positive samples also tested positive for dengue IgG. CONCLUSION: Despite the high specificity of the COVID-19 RDT, we observed cross-reactions and false-positive results between dengue and COVID-19. Dengue and COVID-19 co-infection was also found. Health practitioners in dengue endemic areas should be careful when using antibody RDT for the diagnosis of dengue during the COVID-19 pandemic to avoid misdiagnosis.

Serological cross-reaction and coinfection of dengue and COVID-19 in Asia: Experience from Indonesia.

Similar symptoms and laboratory findings between dengue and coronavirus disease 2019 (COVID-19) pose a diagnostic challenge in some dengue-endemic countries in Asia. In this study, we reported three cases of suspected COVID-19-dengue coinfection in hospitals of Bali, Indonesia. Serological data demonstrated that patients with positive results for dengue virus (DENV) NS1 antigen and anti-dengue IgM were also reactive to COVID-19 rapid antibody tests, suggesting dengue-COVID-19 coinfection. However, two patients were later confirmed negative for SARS-COV-2 by qRT-PCR, implying a plausible cross-reactivity of anti-dengue and anti-COVID-19 antibodies in the serological test. Coinfection of dengue and COVID-19 was evident in one patient, following confirmation of SARS-COV-2 by qRT-PCR and DENV infection using the NS1 antigen serology test. This case was the first case of dengue and COVID-19 coinfection in Indonesia and revealed possible cross-reactivity between SARS-COV-2 and DENV antibodies based on rapid serological tests. Our study indicates a public health concern regarding COVID-19 and dengue detection in Indonesia as well as in other dengue-endemic countries, and it is important for these nations to manage both pathogens concurrently.

Persistence of long COVID symptoms in COVID-19 survivors worldwide and its potential pathogenesis - A systematic review and meta-analysis.

The study sought to determine the prevalence of persistent long COVID symptoms such as anxiety, depression, dizziness, chest pain, sleep difficulty, palpitations, weight loss, and hair loss among coronavirus disease 2019 (COVID-19) survivors worldwide and to discuss the potential pathogeneses. Potential studies were searched in three databases (PubMed, Scopus, and Web of Science) as of January 30, 2021. Data on study characteristics, patient characteristics during the follow-up, the number of patients with persistent long COVID symptoms and total COVID-19 survivors were collected according to PRISMA guidelines. To assess the quality of studies, the Newcastle-Ottawa scale was used. The estimated prevalence of each long COVID symptom and the association between COVID-19 severity and the occurrence of prolonged symptoms was assessed, if appropriate. The global prevalence of prolonged anxiety was 15.76% (95%CI: 6.36%, 25.15%). Chest pain persisted in 10.36% (239/3,224) of COVID-19 patients (95%CI: 4.92%, 15.80%). Prolonged depression was found in 24 of 548 COVID-19 survivors with an estimated prevalence of 4.32% (95%CI: 2.62%, 6.03%) and dizziness was presented in 4.83% (118/2,219, 95%CI: 1.50%, 8.16%) after recovery. Hair loss was complained by 527 of 2,251 recovered patients (cumulative prevalence of 24.76%, 95%CI: 19.60%, 29.91%), while weight loss was identified in 37 cases among 452 COVID-19 survivors (8.19%, 95%CI: 5.66%, 10.71%). Prolonged palpitation was experienced by 19.38% (211/1,926) survivors with 95%CI: 2.40%, 41.16%. Sleep difficulty was found in 541 of 2,622 COVID-19 survivors (17.87%, 95%CI: 7.55%, 28.20%). The association between COVID-19 severity and the occurrence of persistent long COVID symptoms was not analyzed due to the lack of data. In conclusion, persistent psychological symptoms are frequently reported among COVID-19 survivors. Follow-up studies with a longer duration and larger population are warranted to assess the extent of prolonged symptoms and the quality of life of COVID-19 survivors. Despite various potential pathogeneses that have been hypothesized, a definitive mechanism is yet to be addressed. PROSPERO registration: CRD42021247172.

Correlation of miR-150, hsa-let-7e, and miR- 146a and gene expression of IL-6, IL-8, IP-10, and MIP-1ß during dengue virus infection.

Growing evidence suggests that microRNAs (miRNAs) play a pivotal role in viral infection. The objective of this study was to assess the association between the expression of miR- 150, hsa-let-7e, and miR-146a on cytokine expression during dengue infection. Dengue virus (DENV) strain SJN-006, a serotype 2 DENV strain of the Cosmopolitan genotype, isolated in Bali, Indonesia, was used to infect peripheral blood mononuclear cells (PBMCs) isolated from healthy individuals. The relative gene expressions of miR-150, hsa-let-7e, and miR-146a as well as the gene expression of cytokines (IL-6, IL-8, IP-10, and MIP-1ß) were determined using quantitative real time - polymerase chain reaction (qRT-PCR) at 6, 12 and 24 hours post infection (hpi). Correlations between the microRNAs and cytokines were analyzed by means of causality tests. Our data suggests that miR-150 and hsa-let-7e were significantly higher in infected-PBMCs after 12 hpi compared to the uninfected-PBMCs (p<0.05). The causality tests demonstrated that miR-150 and has-let- 7e were negatively correlated with IL-8 expression, meanwhile miR-146a was the contrast. DENV infection was negatively and positively correlated with miR-150 and hsa-let-7e, respectively, after 24 hpi. In conclusion, our data demonstrates the vital role of miR-150, hsa-let-7e, and miR-146a in regulating IL-8 expression with possible different pathways.

Japanese encephalitis virus infection in non-encephalitic acute febrile illness patients.

Although Japanese encephalitis virus (JEV) is considered endemic in Indonesia, there are only limited reports of JEV infection from a small number of geographic areas within the country with the majority of these being neuroinvasive disease cases. Here, we report cases of JEV infection in non-encephalitic acute febrile illness patients from Bali, Indonesia. Paired admission (S1) and discharge (S2) serum specimens from 144 acute febrile illness patients (without evidence of acute dengue virus infection) were retrospectively tested for anti-JEV IgM antibody and confirmed by plaque reduction neutralization test (PRNT) for JEV infection. Twenty-six (18.1%) patients were anti-JEV IgM-positive or equivocal in their S2 specimens, of which 5 (3.5%) and 8 (5.6%) patients met the criteria for confirmed and probable JEV infection, respectively, based on PRNT results. Notably, these non-encephalitic JE cases were less likely to have thrombocytopenia, leukopenia, and lower hematocrit compared with confirmed dengue cases of the same cohort. These findings highlight the need to consider JEV in the diagnostic algorithm for acute febrile illnesses in endemic areas and suggest that JEV as a cause of non-encephalitic disease has likely been underestimated in Indonesia.

Global prevalence and pathogenesis of headache in COVID-19: A systematic review and meta-analysis.

Background: This study was conducted to determine the prevalence of headache in coronavirus disease 2019 (COVID-19) and to assess its association as a predictor for COVID-19. This study also aimed to discuss the possible pathogenesis of headache in COVID-19. Methods: Available articles from PubMed, Scopus, and Web of Science were searched as of September 2 nd, 2020. Data on characteristics of the study, headache and COVID-19 were extracted following the PRISMA guidelines. Biases were assessed using the Newcastle-Ottawa scale. The cumulative prevalence of headache was calculated for the general population (i.e. adults and children). The pooled odd ratio (OR) with 95% confidence intervals (95%CI) was calculated using the Z test to assess the association between headache and the presence of COVID-19 cases. Results: We included 104,751 COVID-19 cases from 78 eligible studies to calculate the global prevalence of headache in COVID-19 and 17 studies were included to calculate the association of headache and COVID-19. The cumulative prevalence of headache in COVID-19 was 25.2% (26,464 out of 104,751 cases). Headache was found to be more prevalent, approximately by two-fold, in COVID-19 patients than in non-COVID-19 patients with symptoms of other respiratory viral infections, OR: 1.73; 95% CI: 1.94, 2.5 with p=0.04. Conclusion: Headache is common among COVID-19 patients and seems to be more common in COVID-19 patients compared to those with the non-COVID-19 viral infection. No definitive mechanisms on how headache  emerges in COVID-19 patients but several possible hypotheses have been proposed. However, extensive studies are warranted to elucidate the mechanisms. PROSPERO registration: CRD42020210332 (28/09/2020).

Predictors of COVID-19 severity: a systematic review and meta-analysis.

Background: The unpredictability of the progression of coronavirus disease 2019 (COVID-19) may be attributed to the low precision of the tools used to predict the prognosis of this disease. Objective: To identify the predictors associated with poor clinical outcomes in patients with COVID-19. Methods: Relevant articles from PubMed, Embase, Cochrane, and Web of Science were searched as of April 5, 2020. The quality of the included papers was appraised using the Newcastle-Ottawa scale (NOS). Data of interest were collected and evaluated for their compatibility for the meta-analysis. Cumulative calculations to determine the correlation and effect estimates were performed using the Z test. Results: In total, 19 papers recording 1,934 mild and 1,644 severe cases of COVID-19 were included. Based on the initial evaluation, 62 potential risk factors were identified for the meta-analysis. Several comorbidities, including chronic respiratory disease, cardiovascular disease, diabetes mellitus, and hypertension were observed more frequent among patients with severe COVID-19 than with the mild ones. Compared to the mild form, severe COVID-19 was associated with symptoms such as dyspnea, anorexia, fatigue, increased respiratory rate, and high systolic blood pressure. Lower levels of lymphocytes and hemoglobin; elevated levels of leukocytes, aspartate aminotransferase, alanine aminotransferase, blood creatinine, blood urea nitrogen, high-sensitivity troponin, creatine kinase, high-sensitivity C-reactive protein, interleukin 6, D-dimer, ferritin, lactate dehydrogenase, and procalcitonin; and a high erythrocyte sedimentation rate were also associated with severe COVID-19. Conclusion: More than 30 risk factors are associated with a higher risk of severe COVID-19. These may serve as useful baseline parameters in the development of prediction tools for COVID-19 prognosis.

Concurrent infections of dengue virus serotypes in Bali, Indonesia.

OBJECTIVE: To describe cases of dengue virus (DENV) concurrent infections in patients from both local and international traveler visiting Bali, Indonesia. RESULTS: During a hospital-based study, 260 patients (from 161 local and 99 international traveler patients) were recruited. Among them, 190 were positive by DENV RT-PCR in which eight patients (five local and three international travelers) detected as having concurrent infections by two different DENV serotypes. Among the eight patients, the common dengue symptoms diagnosed were fever, headache, and myalgia. Six cases (75%) were diagnosed with dengue fever (DF) while two cases (25%) manifested with bleeding and were diagnosed with dengue hemorrhagic fever (DHF) grade 1. The DENVs concurrent infections involved all four DENV serotypes known to be circulating in Bali. Although cases of DENV concurrent infections have been implicated with severe manifestation, we observed that most of concurrent infections cases in our study were of mild clinical manifestation, that may be related to the changing of DENV serotype predominance which is occurring in Bali, Indonesia.