BACKGROUND Hepatitis B virus (HBV) remains difficult to eradicate due to the persistence of covalently closed circular DNA (cccDNA). Matrix metalloproteinase-9 (MMP-9) enhances HBV replication, but the effects of its inhibition remain unexplored. This study aimed to investigate the effects of MMP-9 inhibitors on HBV replication markers. METHODS Primary hepatocyte cultures were obtained from the livers of 6 Tupaia javanica. Cultures were infected with HBV from human sera and divided into control and intervention groups. The intervention group received MMP-9 inhibitors at 1, 3, and 7 nM. The control group received phosphate-buffered saline. Levels of hepatitis B surface antigen (HBsAg), HBV DNA, cccDNA, MMP-9, interferon alpha and beta receptor subunit 1 (IFNAR1), and interferon beta (IFN-β) were measured in both groups before and 72 hours post-intervention. RESULTS MMP-9 inhibitor administration at 1, 3, and 7 nM consistently reduced HBsAg, HBV DNA, cccDNA, and MMP-9 levels, though not statistically significant. Median HBV DNA levels at 1, 3, and 7 nM were 7.05, 5.29, and 5.98 ×103 copies/ml, respectively. Mean cccDNA levels at 1, 3, and 7 nM were 14.15, 11.04, and 13.94 ×103 copies/ml, respectively. The 3 nM dose increased IFNAR1 levels, while the 7 nM dose increased IFN-β, but neither change was significant. Among the tested doses, 3 nM showed the most favorable effects despite the lack of significance. CONCLUSIONS MMP-9 inhibitor suppressed HBsAg, HBV DNA, cccDNA, and MMP-9 while increasing IFNAR1 and IFN-β in vitro.

Purpose: To evaluate the correlation between dry eye symptoms and coronavirus disease 2019 (COVID-19) infection and to assess the real-time reverse transcription–polymerase chain reaction (RT‒PCR) of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) from the conjunctival swab. Methods: A prospective observational case series study was conducted of all suspected and confirmed COVID-19 patients from Dr. Cipto Mangunkusumo Hospital (RSCM) and the Universitas Indonesia Hospital (RSUI). On the first day of the visit (day 0), systemic clinical symptoms and naso-oropharyngeal (NO) RT‒PCR results will classify all subjects as non-, suspected, or confirmed (mild, moderate, and severe) COVID-19. In all patients, we determined the dry eye symptoms based on the Ocular Surface Disease Index (OSDI) and followed up 7(day 7) and 14 days (day 14) after the first visit. When it was technically possible, we also examined the objective dry eye measurements: tear meniscus height (TMH), noninvasive Keratograph® break-up time (NIKBUT), and ocular redness. Additionally, we took conjunctival swab samples for SARS-CoV-2 RT-PCR in all patients. Results: The OSDI scores for 157 patients decreased across days 0, 7, and 14 (median (interquartile range): 2.3 (0–8), 0 (0–3), and 0 (0-0), p value < 0.0001 (D0 vs D14). The moderate-severe COVID-19 group had a higher OSDI score than the other groups at median D0 (15.6 vs 0–2.3), p value < 0.0001 and this pattern was consistently seen at follow-up D7 and D14. However, dry eye complaints were not correlated with the three objective dry eye measurements in mild-moderate COVID-19 patients. NO RT‒PCR results were positive in 32 (20.4%) patients, namely, 13 and 19 moderate-severe and mild COVID-19 patients, respectively. Positive RT‒PCR results were observed in 7/157 (4.5%) conjunctival swab samples from 1 in non-COVID-19 group and 6 in mild group. Conclusion: In the early phase of infection, COVID-19 patients experience dry eye symptoms, which have no correlation with objective dry eye measurements. SARS-CoV-2 in conjunctival swab samples can be detected in patients with normal-to-mild COVID-19, which shows the risk of ocular transmission.

BACKGROUND:Diagnosis for HIV in infants is hard to determine, particularly in limited-resource areas. A delay in the diagnosis of HIV-infected infants will lead to high morbidity and mortality. The purpose of this project is to construct a model of an HIV-positive infant and develop a useful and practical scoring system to estimate the likelihood of mother-to-child transmission that can be applied in the field. METHODS:A cross-sectional study on 100 subjects through medical records of infants born to HIV-infected mothers was conducted at four hospitals and one community health center. Several models of risk prediction scores of HIV-infected infants were then made. Furthermore, the performed validation was performed on 20 subjects of infants born to mothers with HIV in three hospitals by comparing the scoring system and the result of the PCR RNA examination performed at the age of 6 weeks old. RESULTS:The risk of HIV-infected infants was higher in mothers who did not receive ARV through PMTCT programs (OR 33.6; 95% CI 4.0 to 282.2), pulmonary TB infection (OR 5.1; IK95% 1.6 to 16.0) and vaginal delivery (OR 9.2; IK95 2.2 to 38.0%). Two models can predict the occurrence of infected HIV infants effectively. Model 1 consists of maternal age, maternal ARVs, lung TB infection, gestational age, mode of delivery, and sex of the infants with sensitivity and specificity of 78.9% and 70.8% (AUC=0.817 [95% CI 0.709 to 0.926]) and likelihood ratio score of 4. Model 2 consists of ARVs to the mother, pulmonary TB infection, and mode of delivery with sensitivity and specificity of 73.7% and 86.1%; AUC value of 0.812 (95% CI 0.687 to 0.938) and likelihood ratio of 5. External Validation gave similar results to the Model 2 scoring system with PCR RNA. CONCLUSION:The prediction score of HIV-infected infants in Model 2 can be used in newborns of HIV-positive mothers as an effective and practical risk screening tool for HIV-infected infants before the gold standard examination by PCR.

Background and Objectives: Pseudomonas aeruginosa, drug-resistant, causes health infections. Resistance to the preferred therapy meropenem is a serious threat. This study aimed to analyze changes in meropenem minimum inhibitory concentra- tion (MIC), changes in ampC, mexA, and oprD gene expression, and the correlation between MIC and ampC, mexA, and oprD gene expression after meropenem exposure. Materials and Methods: Ten isolates of P. aeruginosa from the Clinical Microbiology Department, Faculty of Medicine, Universitas Indonesia were used. After the bacteria were shown to be sensitive to meropenem phenotypically, intrinsic resis- tance genes were detected using PCR. After meropenem exposure on Days 5 and 12, sensitivity testing was carried out with the concentration gradient method and RNA was detected using real-time RT-PCR. Results: All P. aeruginosa isolates that were phenotypically sensitive to meropenem had the ampC, mexA, and oprD genes. An increase in MIC, an increase in ampC and mexA gene expression, and a decrease in oprD gene expression were observed after meropenem exposure. There was a very strong and significant correlation (p ≤ 0.05) between MIC and oprD gene ex- pression after Day 12 of meropenem exposure. Conclusion: Although there were no significant differences in MIC and ampC, mexA, and oprD gene expression between Day 5 and Day 12, there was a very strong and significant correlation between MIC and oprD gene expression on Day 12 (p ≤ 0.05). This indicates that decreasing oprD gene expression has the potential to increase meropenem resistance in Pseudo- monas aeruginosa.

The current naso-oropharyngeal swab for SARS-CoV-2 detection faces several problems, such as waste issues and its use for quantitative studies. This study aimed to evaluate the total RNA and viral loads from different upper respiratory tract swabs types and whether SARS-CoV-2 quantification can use the current internal control for normalization. This cross-sectional study collected positive specimens with single oropharyngeal or nasopharyngeal swabs and naso-oropharyngeal swabs. The samples were extracted, tested with qualitative RT‒PCR, and then tested with quantitative RT‒PCR. The RNA eluate was measured for the total RNA concentration. The total RNA concentration, viral load, and RNaseP Ct values were collected and analysed statistically. The positive results came from 41 oropharyngeal swabs, 34 nasopharyngeal swabs, and 36 naso-oropharyngeal swabs. The total RNA increased significantly from oropharyngeal swabs to nasopharyngeal swabs to naso-oropharyngeal swabs. Significant differences in RNaseP Ct values between groups and their correlations with total RNA were found. In addition, the increase in the total RNA and the RNaseP Ct values were unrelated to the viral load. The physical features in the naso-oropharyngeal area and the swabbing procedures could affect the total RNA but not the viral load. However, since the virus particles could present inside and outside human cells, the increase in collected human cells may not always be followed by the viral load increase. Normalization using the RNaseP Ct value became unnecessary due to the factors mentioned above. Therefore, a careful approach is needed in viral load studies of swab specimens.

Antimicrobial resistance in bacteria has become a leading global public health issue. Staphylococcus sp. has an efficient mechanism to deal with antimicrobial agents that make them hard to treat in hospital-acquired and community-acquired infections. This study was conducted due to limited data about multidrug resistance and extensively drug resistance in Staphylococcus sp. in Indonesia. This study was a descriptive retrospective study using a cross-sectional design to get the prevalence and antimicrobial susceptibility of S. haemolyticus, S. aureus, and S. epidermidis. The data was secondary data extracted from WHONET 2022 software. This study’s data were from bacteria from samples sent to UKK LMK FKUI, Jakarta from 2017 to 2021 for routine diagnostic. In this study, we found that the prevalence of methicillin-resistant S.aureus was 24,9%, methicillin-resistant S.epidermidis was 65,5%, and methicillin-resistant S.haemolyticus was 86,8%. The prevalence of MDR S.aureus is less than S.epidermidis and S.haemolyticus, respectively. MDR S.haemolyticus was consistently above 85% each year, while S.epidermidis was above 50% and S.aureus was below 50%. XDR Staphylococcus was only found in S.aureus and S.haemolyticus, i.e. three and seven XDR isolates of S.aureus and S.haemolyticus respectively during 2017-2021. Although we could not find any pan-resistant isolates from all samples, we found methicillin-resistant S.aureus and S.haemolyticus isolates that were also resistant to vancomycin and linezolid. S.haemolyticus dan S. epidermidis were an important coagulase-negative Staphylococcus species that can’t be neglected due to the high percentage of MDR and the discoveries of XDR in S.haemolyticus so that they have the potential to disseminate resistance plasmids to the more virulent bacteria. Therefore we need to control the use of antimicrobial agent to prevent this resistance.

Background: SARS-CoV-2 rapid mutation generates many concerning new strains. Although lockdown had been applied to contain the disease, the household remains a critical place for its transmission. This study aimed to assess the variation of SARS-CoV-2 strains and their clinical manifestations within family clusters in Jakarta, Indonesia. Method: Naso-oropharyngeal swab specimens from family clusters positive for SARS-CoV-2 were collected for whole-genome sequencing. Their baseline data, symptoms, and source of infection were recorded. The whole-genome data was then analyzed with the bioinformatics program to evaluate the SARS-CoV-2 genome characteristic and submitted to GISAID for strain identification. The phylogenetic tree was built to observe the relationship between virus strain within the family cluster and its clinical manifestation. Result: This study obtained eight family clusters from twenty-two patients. Half of the cluster's source of infection was a family member who had to work at the office. The infection rate ranged from 37.5% to 100%. The phylogenetic tree showed that the same virus within a cluster could generate various clinical outcomes. Five clusters had one family member with pneumonia, while others had mild to no symptoms. Four breakthrough cases were detected in this study, which were infected by the virus from GH and GK clade. Conclusion: Our study observed the SARS-CoV-2 transmission to the household setting through the workplace, which might become a common pathway if the pandemic status is removed. Although vaccination is expected to reduce the burden of COVID-19, adequate control measures remain essential, given that breakthrough infections are evident.

Objective: This study aims to evaluate the effect of Clostridium perfringens sialidase treatment on monolayer cell behavior using computational screening and an in vitro approach to demonstrate interaction between enzyme-based drugs and ligands in host cells. Materials and Methods: The in silico study was carried out by molecular docking analysis used to predict the interactions between atoms that occur, followed by genetic characterization of sialidase from a wild isolate. Sialidase, which has undergone further production and purification processes exposed to chicken embryonic fibroblast cell culture, and observations-based structural morphology of cells compared between treated cells and normal cells without treatment. Results: Based on an in silico study, C. perfringens sialidase has an excellent binding affinity with Neu5Acα (2.3) Gal ligand receptor with Gibbs energy value (ΔG)—7.35 kcal/mol and Ki value of 4.11 μM. Wild C. perfringens isolates in this study have 99.1%–100% similarity to the plc gene, NanH, and NanI genes, while NanJ shows 93.18% similarity compared to the reference isolate from GenBank. Sialidase at 750 and 150 mU may impact the viability, cell count, and cell behavior structure of fibroblast cells by significantly increasing the empty area and perimeter of chicken embryo fibroblast (CEF) cells, while at 30 mU sialidase shows no significant difference compared with mock control. Conclusion: Sialidase-derived C. perfringens has the capacity to compete with viral molecules for attachment to host sialic acid based on in silico analysis. However, sialidase treatment has an impact on monolayer cell fibroblasts given exposure to high doses.

Background: The M protein is one of the structural protein of SARS CoV-2. The M protein is relatively conserved and stable than other structural protein. This made immunodominant epitopes of M protein has the advantage to be learned with the aim to understanding its immunogenicity and its antigenicity. Several studies have shown that M protein successfully expressed in several expression system, E. coli was one of them. In this study, M gene of SARS CoV-2 was cloned, sequenced, expressed in E. coli BL21 (DE3) system, and purified with denature condition. The membrane recombinant protein can be used for development of a SARS CoV-2 antibody diagnostic system.Methods: the gene encoding the SARS-CoV-2 M protein in the form of gBlocks was cloned into the cloning vector and then subcloned into the pQE80L prokaryotic expression vector. There were three stages of recombinant plasmids verification which were the colony PCR, restriction, and sequencing. The M gene cloned in pQE80L was expressed by using BL21 and purified under denature condition.Results: The recombinant plasmid pQE80L was confirmed containing M protein using primer that specifically amplify the multiple cloning sites (MCS) of pQE80L and produce 595 bp amplicon that indicating the presence of recombinant gene. Restriction of recombinant plasmid using BamHI and HindIII produced 306 bp and 4709 bp DNA bands. The sequence of M gene in pEQ80L has been confirmed by sequencing. Further to ensure the M gene could be expressed in prokaryotic system, the recombinant plasmid was transformed into BL21 bacteria. The SARS-CoV-2 membrane protein with a size of 11,83 kDa has been successfully expressed and purified using the Ni-NTA agarose purification technique under denature conditions.Conclusion: the gene encoding the membrane protein of SARS-CoV-2 has been successfully cloned and expressed in the prokaryotic expression system. AbstrakLatar belakang: Protein M merupakan salah satu protein struktural SARS-CoV-2. Protein M merupakan protein yang relatif lestari dan tidak mudah untuk bermutasi dibandingkan protein struktural lainnya. Hal ini menjadikan protein M memiliki kelebihan untuk dipelajari epitop imunodominannya, dengan tujuan untuk memfasilitasi pemahaman terkait imunogenisitas dan antigenisitasnya. Beberapa penelitian menunjukkan ekspresi protein M telah dilakukan pada beberapa sistem ekspresi, salah satunya pada sistem ekspresi E. coli. Pada penelitian ini dilakukan pengklonaan gen M SARS CoV-2, sekuensing, ekspresi pada E. coli BL21(DE3), dan purifikasi dengan kondisi denatur. Protein rekombinan membran ini kemudian dapat digunakan dalam pengembangan sistem diagnostik antibodi SARS CoV-2.Metode: gen penyandi protein M SARS-CoV-2 dalam bentuk gBlocks diklona ke dalam vector pengklonaan dan kemudian disubklona ke dalam vector ekspresi prokariot pQE80L. Tiga tahapan verifikasi terhadap plasmid rekombinan, yaitu PCR koloni, restriksi, dan sekuensing dilakukan untuk memastikan bahwa gen target telah berhasil diklona. Plasmid ekspresi pQE80L yang membawa gen M kemudian diekspresikan dan dipurifikasi pada kondisi denatur. Hasil: Plasmid rekombinan pQE80L dikonfirmasi mengandung protein M menggunakan primer yang secara spesifik mengamplifikasi multiple cloning site (MCS) pQE80L dan menghasilkan amplicon berukuran 595 bp yang menunjukkan keberadaan gen rekombinan. Restriksi plasmid rekombinan menggunakan BamHI dan HindIII menghasilkan pita DNA berukuran 306 bp dan 4709 bp. Sekuen genM di pQE80L dikonfirmasi dengan sekuensing. Untuk memastikan bahwa gen M dapat diekspresikan di sistem prokariot, plasmid rekombinan ditransformasi ke dalam bakteri BL21. Protein membran SARSCoV-2 dengan ukuran 11,83 kDa telah berhasil diekspresikan dan dipurifikasi menggunakan teknik purifikasi Ni-NTA agarose pada kondisi denature.Kesimpulan: gen penyandi protein membrane SARS-CoV-2 telah berhasil diklona dan diekspresikan pada sistem ekspresi prokariot

Background: The gut microbiota plays an important role in maintaining gut health and is linked to various intestinal disorders. Antibiotic use for hospitalized patients can have several negative effects on the gut microbiota, including reduced species diversity, altered metabolic activity, and the development of antibiotic resistance. Dysbiosis of gut microbiota corresponded with dominant microbiota will trigger diarrhea in both adults and children.Methods: This retrospective study used a cross-sectional approach to determine the profile of the dominant gut microbiota. Stool specimens from hospitalized patients with complaints of diarrhea were examined at Clinical Microbiology Laboratory of FMUI Jakarta during the period 2018 to 2022. The specimens were inoculated on SS and EMB agar. Selenite Broth was used as an enrichment medium for the isolation of salmonellas. The media were then incubated at 35 degrees-37 degrees C for 18-24 hours. The dominant colony was identified using Gram staining and VITEK2 compact system (bioMerieux (R)).Results: A total of 367 stool specimens, of which only 325 samples were included in the study. Two microorganisms were found most in adult and pediatric patients, Escherichia coli and Klebsiella pneumoniae were 119 (43.90%) and 56 (20.66%) respectively. The third most common microorganism was found to be different between adult and pediatric patients. The third dominant microbiota in pediatric patients were Enterobacter cloacae, while in adults were Canaiaa albicans, 23 (8.49%) and 18 (6.64%) respectively.Conclusion: Overall, the dominant microbiota in children and adults is E. coli and K. pneumoniae. The third dominant microbiota in pediatric patients were E. cloacae, while in adults were C. albicans. The differences in intestinal microbiota profiles in adult and pediatric patients found in this study provide new insights into guidelines for the management of diarrhea in hospitalized adults and children.

Diagnosis of Neisseria gonorrhoeae infection is needed for patient therapy and for reducing this bacterial transmission in the population. The culture method is a gold standard method for N. gonorrhoeae detection, however it has low sensitivity. Among molecular methods with high sensitivity and specificity, SYBR Green real-time PCR is the potential method for N. gonorrhoeae detection. In this study, we developed an SYBR Green real-time PCR-based system assay for N. gonorrhoeae detection. Several PCR conditions were optimized and analyzed including primer annealing temperature, DNA template volume, the limit of detection (LoD), cross-reaction with others (bacteria, viruses, fungus, protozoa), and quality assurance. The results showed that the annealing temperature and DNA template volume were 60oC and 5 µL, respectively. The LoD was 29 DNA copies corresponding to 3 bacterial cells per reaction. No cross-reaction was detected for other bacteria, viruses, fungus and protozoa. The external quality assurances enrolled in 2019 and 2021 showed 100% concordance. The preliminary testing for clinical samples was also 100% concordance. In conclusion, the SYBR Green real-time PCR-based system assay developed in this study is promising for application in clinical laboratories.

Background and Aim: Clostridium toxins are widely used as medicinal agents. Many active metabolic enzymes, including sialidase (neuraminidase), hyaluronidase, and collagenase, contribute to the mechanism of action of these toxins. Sialidase from Clostridium perfringens recognizes and degrades sialic acid receptors in the host cell glycoprotein, glycolipid, and polysaccharide complexes. Sialic acid promotes the adhesion of various pathogens, including viruses, under pathological conditions. This study aimed to investigate the potential of C. perfringens sialidase protein to inhibit Newcastle disease virus (NDV) infection in ovo model. Materials and Methods: C. perfringens was characterized by molecular identification through polymerase chain reaction (PCR) and is cultured in a broth medium to produce sialidase. In addition, sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis was conducted to characterize the sialidase protein. In contrast, enzymatic activity and protein concentration were carried out using a neuraminidase assay kit and Bradford to obtain suitable active substances. Furthermore, embryonated chicken egg models were used to observe the toxicity of several sialidase doses. Then, the hemagglutination (HA) titer was obtained, and absolute quantitative reverse transcription–PCR assay was performed to measure the viral replication inhibitory activity of sialidase against NDV. Results: Each isolate had a specific sialidase gene and its product. The sialidase derived from C. perfringens could hydrolyze the sialic acid receptor Neu5Ac (2,6)-Gal higher than Neu5Ac (2,3)Gal in chicken erythrocytes, as observed by enzyme-linked lectin assay. A significant difference (p = 0.05) in the HA titer in the pre-challenge administration group at dosages of 375 mU, 187.5 mU, and 93.75 mU in the competitive inhibition experiment suggests that sialidase inhibits NDV reproduction. Quantification of infective viral copy confirmed the interference of viral replication in the pre-challenge administration group, with a significant difference (p = 0.05) at the treatment doses of 750 mU, 375 mU, and 46.87 mU. Conclusion: The potency of sialidase obtained from C. perfringens was shown in this study, given its ability to reduce the viral titer and copy number in allantoic fluids without adversely impacting the toxicity of the chicken embryo at different concentrations.

Lower respiratory tract, sepsis, or urinary tract infection caused by the multidrug resistance Pseudomonas aeruginosa is common in the hospital, especially in the ICU wards. The treatment against this bacteria requires combination of antibiotics with different mechanism of actions. In this study, several combinations of antibiotics were evaluated in vitro against carbapenem-resistant P. aeruginosa isolated from the ICU of Cipto Mangunkusumo Hospital. The combination of antibiotics tested were ceftazidime-amikacin, ceftazidime-ciprofloxacin, and ciprofloxacin-amikacin. Checkerboard assay to the combination of antibiotics was conducted to assess the in vitro synergistic activity. A total of 22 P. aeruginosa isolates were collected, 16 of them were resistant to ceftazidime, ciprofloxacin, amikacin, as well as carbapenem. The result revealed that the combination of ceftazidime and amikacin showed promising synergistic activity. Conversely, no synergitic activities were shown by the combination of ceftazidime-ciprofloxacin and ciprofloxacin-amikacin. The combination of ceftazidime-amikacin may has potential effect againsts carbapenem-resistant P. aeruginosa in vitro.

Objective: The Newcastle disease virus (NDV) is an infectious disease that causes very high eco¬nomic losses due to decreased livestock production and poultry deaths. The vaccine’s ineffec¬tiveness due to mutation of the genetic structure of the virus impacts obstacles in controlling the disease, especially in some endemic areas. This study aimed to provide an alternative treatment for NDV infection by observing the viral replication inhibitor activity of Clostridium perfringens sialidase in primary chicken embryo fibroblast (CEF) cells. Materials and Methods: The virus was adapted in CEF monolayer cells, then collected thrice using the freeze–thaw method and stored at −20°C for the next step in the challenge procedure. C. perfringens crude sialidase was obtained, but it was further purified via stepwise elution in ion exchange using Q Sepharose® Fast Flow and affinity chromatography with oxamic acid agarose. The purified sialidase was tested for its toxicity, ability to breakdown sialic acid, stopping viral replication, and how treated cells expressed their genes. Results: According to this study, purified C. perfringens sialidase at dosages of 187.5, 93.75, and 46.87 mU effectively hydrolyzes CEF cells’ sialic acid and significantly inhibits viral replication on the treated cells. However, sialidase dosages of 375 and 750 mU affected the viability of mono¬layer CEF cells. Interestingly, downregulation of toll-like receptor (TLR)3 and TLR7 (p < 0.05) in the sialidase-treated group indicates viral endocytosis failure. Conclusions: By stopping endocytosis and viral replication in host cells, sialidase from C. perfrin¬gens can be used as an alternative preventive treatment for NDV infection.

BACKGROUND The B.1.617.2 (delta) variant of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first discovered in Maharashtra in late 2020 and has rapidly expanded across India and worldwide. It took only 2 mo for this variant to spread in Indonesia, making the country the new epicenter of the delta variant as of July 2021. Despite efforts made by accelerating massive rollouts of current vaccines to protect against infection, cases of fully-vaccinated people infected with the delta variant have been reported. AIM To describe the demographic statistics and clinical presentation of the delta variant infection after the second dose of vaccine in Indonesia. METHODS A retrospective, single-centre case series of the general consecutive population that worked or studied at Faculty of Medicine, Universitas Indonesia with confirmed Delta Variant Infection after a second dose of vaccine from 24 June and 25 June 2021. Cases were collected retrospectively based on a combination of author recall, reverse transcription-polymerase chain reaction (RT-PCR), and whole genome sequencing results from the Clinical Microbiology Laboratory, Faculty of Medicine, Universitas Indonesia. RESULTS Between 24 June and 25 June 2021, 15 subjects were confirmed with the B.1.617.2 (delta) variant infection after a second dose of the vaccine. Fourteen subjects were vaccinated with CoronaVac (Sinovac) and one subject with ChAdOx1 nCoV-19 (Oxford-AstraZeneca). All of the subjects remained in home isolation, with fever being the most common symptom at the onset of illness (n = 10, 66.67%). The mean duration of symptoms was 7.73 d (± 5.444). The mean time that elapsed from the first positive swab to a negative RT-PCR test for SARS-CoV-2 was 17.93 d (± 6.3464). The median time that elapsed from the second dose of vaccine to the first positive swab was 87 d (interquartile range: 86-128). CONCLUSION Although this case shows that after two doses of vaccine, subjects are still susceptible to the delta variant infection, currently available vaccines remain the most effective protection. They reduce clinical manifestations of COVID-19, decrease recovery time from the first positive swab to negative swab, and lower the probability of hospitalization and mortality rate compared to unvaccinated individuals.

COVID-19 is a disease caused by SARS-CoV-2, a new virus from genus β-coronaviruses. This disease has been declared a pandemic by WHO on 11 March 2020 until now. The nucleic acid tests are the most frequently used assays because of their high sensitivity and specificity. One of the tests is the GeneXpert, a real-time reverse transcription polymerase chain reaction (rRT-PCR)-based assay platform. The use of the GeneXpert shows great public health interest because of the rapid (50 min), the minimum number of trained staff, and less infrastructure and equipment. However, there are limited data on the application of the GeneXpert for the detection of SARS-CoV-2. Therefore, we conducted a comparative study between the GeneXpert and in-house N1N2 CDC rRT-PCR assay. Of 86 samples, 17 were rRT-PCR positive while 13 were GeneXpert positive. Of rRT-PCR positive 17 samples, 7 were GeneXpert negative [58.82% (10/17] sensitivity]. We also found that 3 GeneXpert positive samples showed rRT-PCR negative (95.65% [66/69] specificity). It is concluded that negative results by the GeneXpert can not rule out the possibility of SARS-CoV-2 infection, particularly in close-contact individuals and the interpretation of the positive result should be analyzed carefully, particularly amplification with Ct>40.

Objective: This research investigates the antibacterial, anticancer, antioxidant, and antiretroviral activities of the lionfish spine poison extract. Methods: Isolation and purification of the phospholipase A2 (PLA2) protein obtained from the spine poison were conducted through the following stages, including, extraction of the venom by sonication, heating, and purification using gradual saturation levels of ammonium sulfate. Furthermore, the purity and concentration of PLA2 were analyzed using the Lowry test and Marinetti’s method, respectively, while its protein content was ascertained through SDS-PAGE. Toxicity was then evaluated employing the brine shrimp lethality test (BSLT), and its anticancer activity was assessed in human cervical carcinoma cells (HeLa cells). Finally, its antioxidant, antibacterial, and antiretroviral activities were analyzed using the DPPH method, agar diffusion test against Salmonella sp. and E. coli, and SRV-2 and RT-qPCR tests, respectively. Results: The protein demonstrated 37.79% inhibition for anticancer activity, IC50 1312 ppm for antioxidant activity, 98.81%, and 89.28% inhibition of E. coli and Salmonella sp. respectively for antibacterial activity and 98.13% inhibition for antiretroviral activity. Conclusion: It can be concluded that lionfish (Pterois volitans) has the potential to be developed as an antioxidant, anticancer, antibacterial, and antiretroviral agent. Furthermore, the pharmacological activity of its spine venom was determined by isolating PLA2 protein from its extract, using an optimum heating temperature of 70 °C and an ammonium sulfate saturation level of 80%.

of the original source.These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.recipient of red cell unit tested for COVID-19 PCR and all were nonreactive.None of the recipients developed any COVID-19 related symptoms post-transfusion Conclusion: None of the recipients of donor diagnosed with COVID-19 following donation developed COVID-19 related symptoms or tested positive for COVID-19 PCR and remaining blood products were also negative for COVID-19 PCR in this asymptomatic blood donor.Continuous data collection is needed to conclude the possibility of COVID-19 transmission through blood transfusion.

Introduction. The COVID-19 pandemic has entered a new phase with the roll-out of several vaccines worldwide at an accelerated phase. The occurrence of a more severe presentation of COVID-19 after vaccination may affect policymakers' decision-making and vaccine uptake by the public. Vaccine-associated disease enhancement (VADE) is the modified presentation of infections in individuals after having received a prior vaccination. Currently, little is known about the potential of vaccine-associated disease enhancement (VADE) following COVID-19 immunization. Case Illustration. We herewith report two patients admitted with confirmed COVID-19 pneumonia with a history of CoronaVac vaccination. The first patient with a relatively milder course of the disease had received two doses of CoronaVac, whereas the second patient with a more progressive course of the disease received only one dose before developing symptoms and being admitted to the hospital. Our observations suggest that vaccination could act in boosting the inflammatory process and reveal the previously asymptomatic COVID-19 illness. Theoretically, vaccines could induce VADE, where only suboptimal, nonprotective titers of neutralizing antibodies were produced or proinflammatory T-helper type 2 response was induced. Secondly, enhanced respiratory disease (ERD) could manifest, where pulmonary symptoms are more severe due to peribronchial monocytic and eosinophilic infiltration. Understanding VADE is important for the decision-making by the public, clinicians, and policymakers and is warranted for successful vaccination uptake. Conclusion. We report two cases of patients developing COVID-19 shortly after CoronaVac vaccination in which VADE is likely. We recommend that current vaccination strategies consider the measurement of neutralizing antibody titer as a guide in ensuring the safest strategy for mass immunization. Studies are needed to investigate the true incidence of VADE on vaccinated individuals as well as on how to differentiate between VADE and severe manifestations of COVID-19 that are unrelated to vaccination.

Besides human papillomavirus (HPV), the cervical carcinogenesis is also affected by many risk factors including pathogenic bacteria such as Chlamydia trachomatis (CT), Mycoplasma spp. (MS), Ureaplasma urealyticum (UU), and U. parvum (UP) infections. Thus, we studied the bacterial infections for 68 patients with cervical cancerous cases and other cervical problems. Cervical swab samples were collected by specialist doctors and tested for the pathogenic bacteria by real time polymerase chain reaction (rPCR) and HPV by conventional PCR. Of 68 patients, 22 were diagnosed as cervical cancer and 46 were diagnosed as other cervical problems. All patients with cervical cancer were HPV positive, while the patients with other cervical problems were HPV negative. Of 22 HPV positive-cervical cancer patients, 7 (31.82%), 6 (27.27%), 3 (13.64%) were positive for MS, UU, and UP, respectively. None of the patients was CT positive. For 46 HPV negative-other cervical problem patients, 6 (13.04%), 12 (26.09%), 20 (43.48%), and 22 (47.83%) were positive for CT, MS, UU, and UP, respectively. There was no association of CT, MS, and UU infections with the HPV positive-cervical cancer patients (p >0.05). However, there is a negative correlation between UP infection and the HPV negative-other cervical problem patients (p<0.05).