Pratiwi Sudarmono

The burden of diarrhoea, shigellosis, and cholera in North Jakarta, Indonesia: findings from 24 months surveillance

BackgroundIn preparation of vaccines trials to estimate protection against shigellosis and cholera we conducted a two-year community-based surveillance study in an impoverished area of North Jakarta which provided updated information on the disease burden in the area.MethodsWe conducted a two-year community-based surveillance study from August 2001 to July 2003 in an impoverished area of North Jakarta to assess the burden of diarrhoea, shigellosis, and cholera. At participating health care providers, a case report form was completed and stool sample collected from cases presenting with diarrhoea.ResultsInfants had the highest incidences of diarrhoea (759/1 000/year) and cholera (4/1 000/year). Diarrhea incidence was significantly higher in boys under 5 years (387/1 000/year) than girls under 5 years (309/1 000/year; p < 0.001). Children aged 1 to 2 years had the highest incidence of shigellosis (32/1 000/year). Shigella flexneri was the most common Shigella species isolated and 73% to 95% of these isolates were resistant to ampicillin, trimethoprim-sulfamethoxazole, chloramphenicol and tetracycline but remain susceptible to nalidixic acid, ciprofloxacin, and ceftriaxone. We found an overall incidence of cholera of 0.5/1 000/year. Cholera was most common in children, with the highest incidence at 4/1 000/year in those less than 1 year of age. Of the 154 V. cholerae O1 isolates, 89 (58%) were of the El Tor Ogawa serotype and 65 (42%) were El Tor Inaba. Thirty-four percent of patients with cholera were intravenously rehydrated and 22% required hospitalization. V. parahaemolyticus infections were detected sporadically but increased from July 2002 onwards.ConclusionDiarrhoea causes a heavy public health burden in Jakarta particularly in young children. The impact of shigellosis is exacerbated by the threat of antimicrobial resistance, whereas that of cholera is aggravated by its severe manifestations.

Antiviral activity of extracts from Morinda citrifolia leaves and chlorophyll catabolites, pheophorbide a and pyropheophorbide a, against hepatitis C virus

The development of complementary and/or alternative drugs for treatment of hepatitis C virus (HCV) infection is still needed. Antiviral compounds in medicinal plants are potentially good targets to study. Morinda citrifolia is a common plant distributed widely in Indo‐Pacific region; its fruits and leaves are food sources and are also used as a treatment in traditional medicine. In this study, using a HCV cell culture system, it was demonstrated that a methanol extract, its n‐hexane, and ethyl acetate fractions from M. citrifolia leaves possess anti‐HCV activities with 50%‐inhibitory concentrations (IC50) of 20.6, 6.1, and 6.6 μg/mL, respectively. Bioactivity‐guided purification and structural analysis led to isolation and identification of pheophorbide a, the major catabolite of chlorophyll a, as an anti‐HCV compound present in the extracts (IC50  = 0.3 μg/mL). It was also found that pyropheophorbide a possesses anti‐HCV activity (IC50 = 0.2 μg/mL). The 50%‐cytotoxic concentrations (CC50) of pheophorbide a and pyropheophorbide a were 10.0 and 7.2 μg/mL, respectively, their selectivity indexes being 33 and 36, respectively. On the other hand, chlorophyll a, sodium copper chlorophyllin, and pheophytin a barely, or only marginally, exhibited anti‐HCV activities. Time‐of‐addition analysis revealed that pheophorbide a and pyropheophorbide a act at both entry and the post‐entry steps. The present results suggest that pheophorbide a and its related compounds would be good candidates for seed compounds for developing antivirals against HCV.

A multicentre surveillance study on the characteristics, bacterial aetiologies and in vitro antibiotic susceptibilities in patients with acute exacerbations of chronic bronchitis

Background and objective:  Antimicrobial resistance is a global problem and the prevalence is high in many Asian countries.

Induction of cell-mediated immune responses in mice by DNA vaccines that express hepatitis C virus NS3 mutants lacking serine protease and NTPase/RNA helicase activities.

Effective therapeutic vaccines against virus infection must induce sufficient levels of cell-mediated immune responses against the target viral epitopes and also must avoid concomitant risk factors, such as potential carcinogenic properties. The nonstructural protein 3 (NS3) of hepatitis C virus (HCV) carries a variety of CD4+ and CD8+ T cell epitopes, and induces strong HCV-specific T cell responses, which are correlated with viral clearance and resolution of acute HCV infection. On the other hand, NS3 possesses serine protease and nucleoside triphosphatase (NTPase)/RNA helicase activities, which not only play important roles in viral life cycle but also concomitantly interfere with host defense mechanisms by deregulating normal cellular functions. In this study, we constructed a series of DNA vaccines that express NS3 of HCV. To avoid the potential harm of NS3, we introduced mutations to the catalytic triad of the serine protease (H57A, D81A and S139A) and the NTPase/RNA helicase domain (K210N, F444A, R461Q and W501A) to eliminate the enzymatic activities. Immunization of BALB/c mice with each of the DNA vaccine candidates (pNS3[S139A/K210N], pNS3[S139A/F444A], pNS3[S139A/R461Q] and pNS3[S139A/W501A]) that expresses an NS3 mutant lacking both serine protease and NTPase/helicase activities induced T cell immune responses to the degree comparable to that induced by the wild type NS3 and the NS3/4A complex, as demonstrated by interferon-γ production and cytotoxic T lymphocytes activities against NS3. The present study has demonstrated that plasmids expressing NS3 mutants, NS3(S139A/K210N), NS3(S139A/F444A), NS3(S139A/R461Q) and NS3(S139A/W501A), which lack both serine protease and NTPase/RNA helicase activities, would be good candidates for safe and efficient therapeutic DNA vaccines against HCV infection.

Anti-hepatitis C virus activity of a crude extract from longan ( Dimocarpus longan Lour.) leaves

Infection with hepatitis C virus (HCV) results in hepatitis C, a disease characterized by chronic infection, cirrhosis, and hepatocellular carcinoma. Currently, the standard therapy is a combination of pegylated interferon-α plus ribavirin with NS3 protease inhibitors. Addition of NS3 protease inhibitors to the standard therapy improves response rates; however, use of NS3 protease inhibitors is also associated with significant adverse effects and an increase in the overall cost of treatment. Therefore, there is a need to develop safe and inexpensive drugs for the treatment of HCV infections. In this study, we examined the antiviral activity of a crude extract from Dimocarpus longan leaves against HCV (genotype 2a strain JFH1). The D. longan crude extract (DL-CE) exhibited anti-HCV activity with a 50% effective concentration (EC50) of 19.4 μg/ml without cytotoxicity. A time-of-addition study demonstrated that DL-CE has anti-HCV activity at both the entry and post-entry steps and markedly blocks the viral entry step through direct virucidal activity with marginal inhibition of virion assembly. Co-treatment of DL-CE with cyclosporine A, an immunosuppressant or telaprevir, an NS3 protease inhibitor, resulted in additive and synergistic antiviral effects, respectively. Our findings suggest that DL-CE may be useful as an add-on therapy candidate for treating HCV infections.

Immunogenicity of a candidate DNA vaccine based on the prM/E genes of a dengue type 2 virus Cosmopolitan genotype

The development of a dengue virus vaccine is a major priority in efforts to control the diseases. Several researchers are currently using the Asian 1 and Asian 2 genotypes as vaccine candidates for dengue type 2 virus (DENV-2). However, in this study, we constructed a recombinant plasmid-based prM/E gene, from a DENV-2 Cosmopolitan genotype strain as a dengue DNA vaccine candidate. The protein expression of the recombinant plasmid in CHO cells was analyzed using an enzyme-linked immunosorbent assay, western blotting, and sucrose gradient sedimentation. After being used to immunize ddY mice three times at doses of 25 or 100 μg, the DNA vaccine induced humoral immune responses. There was no difference in the neutralizing antibody titer (focus reduction neutralization test 50% value) of mice immunized with 25 and 100 μg DNA vaccine doses. When challenged with 3 × 10(5) FFU DENV-2, immunized mice could raise anamnestic neutralizing antibody responses, which were observed at day 4 and day 8 post-challenge. Analysis of immunogenicity using BALB/c mice showed that their antibody neutralization titers were lower than those of ddY mice. In addition, the antibodies produced after immunization and challenge could also neutralize a DENV-2 Asian 2 genotype (New Guinea C) strain. Therefore, the DENV-2 Cosmopolitan genotype may be a DENV-2 vaccine candidate.

The Etiologies and Clinical Characteristics of Patients Hospitalized with an Acute Febrile Illness and Central Nervous System Sydromes in Indonesia

Abstract Background Acute febrile illness is a common reason for hospitalization in many developing countries, including Indonesia. While patients can often be categorized and managed based on clinical presentations, diagnostic capacity in these countries remains limited, leading to poor patient outcomes. For patients with central nervous system (CNS) infections, identifying the underlying etiologies is particularly important to prevent lifelong neurological complications and death. Methods As part of a study conducted at 8 top-referral hospitals across Indonesia from 2013 to 2016, 114 of 1,486 enrolled subjects presented with an acute fever and a CNS syndrome. To identify the etiologies and clinical manifestations of these infections, as well as the management of febrile patients at the hospitals, demographic and clinical data were collected at enrollment, and blood samples were collected for diagnostic testing at enrollment, once during days 14–28, and at 3 months after enrollment. Results Subject ages ranged from 1 to 63.2 years old (median of 4.9 years old), and underlying diseases were reported in 35 (30.7%) subjects. Standard-of-care, molecular, and serological testing identified pathogens in 56 (49.1%) cases, as detailed in the table. Of the 19 subjects who died, 18 presented with decreased consciousness and 5 were infected with Rickettsia typhi, which was clinically misdiagnosed in each case. Conclusion The findings from this study will improve the diagnosis and treatment of patients presenting with CNS syndromes in Indonesia. Additionally, the discovery of misdiagnosed, fatal etiologies highlights the general need for greater diagnostic testing capacity to aid clinicians and inform public health policy makers. Acute febrile patients with neurological signs and symptoms Consciousness status (n) Normal (61) Decreased (53) Mortality (%) 1.6 34 End-of-study status (n) Discharged (60) Died (1) Discharged (35) Died (18) Etiology (n) Unknown (32) Unknown (1) Unknown (16) Unknown (9) HHV-6 (9) Dengue (5) R. typhi (5) Dengue (8) HHV-6 (3) Dengue (1) Chikungunya (5) Influenza (2) Influenza (1) Influenza (3) E. faecalis (2) Salmonella spp. (1) K. pneumoniae 
(1) Chikungunya 
(1) S. pneumoniae 
(1) Amoeba (1) Leptospira spp. (1) S. Typhi (1) E. coli (1) S. Aureus (1) Seoul Virus (1) S.Typhi (1) RSV (1) Disclosures All authors: No reported disclosures.

INA-RESPOND: a multi-centre clinical research network in Indonesia

Nationally representative observational and translational research is needed to address the public health challenges in Indonesia due to the geographic disparity, recently decentralized health system, and diverse infectious disease priorities. To accomplish this, the Indonesian Ministry of Health in collaboration with the US National Institute of Health has established INA-RESPOND (Indonesia Research Partnership on Infectious Disease) – a clinical research network comprising 9 referral hospitals, 7 medical faculties, and 2 research centres across Indonesia. The network provides a forum to conduct research at a national scale and to address scientific questions that would be difficult to address in smaller research settings. Further, it is currently conducting multi-centre research on the etiologies of fever, sepsis, and tuberculosis. There are opportunities to leverage existing network resources for other public health research needs. INA-RESPOND is an Indonesian-led network in a country with diverse population groups and public health needs which is poised to collaborate with researchers, universities, donors, and industry worldwide. This paper describes the network and its goals and values, as well as the management structure, process for collaboration, and future vision.

STUDI PEMETAAN AWAL DNA Mycobacterium tuberculosis COMPLEX SECARA Spoligotyping PADA HASIL ISOLASI DAHAK PASIEN TUBERKULOSIS PARU DARI 10 IBU KOTA PROPINSI (BAGIAN I)

Abstract. Mapping TB genotype of Mycobacterium tuberculosis (Mtb) is an important study to identify their distribution or characteristic and also may lead to improve control of the disease. This study conducted a preliminary mapping of the tubercle bacilli which had been circulating in Indonesia. Cultures of DNA isolated from TB patients at urban areas in 16 provinces in Indonesia, are chosen based on TB Case Detection Rate (CDR) 2006 from Indonesia Directorate General of Communicable Disease Control and Environment Health (Ministry of Health), were analyzed by spoligotyping for strain differentiation. In this first part, the analyzed result only came from urban areas in 10 provinces, i.e. Palembang, Bandar Lampung, Serang, Jakarta, Bandung, Surabaya, Banjarmasin, Makassar, Pontianak and Ambon. Sample were 269 DNA from 294 isolates collected from sputum of suspect TB patients with sputum-smear positive (SS+) and age above 15 years old. All samples were obtained from peripheral health laboratory in each province. The procedure collection is accordance to Indonesia DOTs guidelines (AFB smears) and samples were transported from those 10 areas to Bacteriology Laboratory at CBPRD. Sputum was taken for culture in liquid media MGIT Bactec 960 and solid media Lowenstein Jensen. The DNAs from positive liquid media MGIT Bactec 960 were isolated and analyzed by spoligotyping to identify the spoligo pattern. The spoligotyping results converted into octal format within Words & Excel spreadsheets and compared to International Spoligotype-database (SpolDB4). The previous study (Parwati et.al.) found some differences geographical distribution between Beijing genotype strain of tubercle bacilli in West Indonesia compare to East Indonesia, and the same pattern was also found in this study. Furthermore, the results in this study showed the differences in spoligo pattern of Mtb complex at 10 urban areas in West, Middle and East Indonesia. The percentage of Beijing strain family in the samples from West Indonesia showed around 26.61% (31.48% in Sumatra, 28.83% in Java and 16.98% in Kalimantan); from the Middle Indonesia around 25.93%; and none were found in samples from the East Indonesia. The SpolDB4 pattern also showed that the majority of isolates belonged to major clades of M.tuberculosis, i.e. the East African-Indian (EAI); Haarlem (H), Latin American-Mediterranean (LAM), the Central and Middle Eastern Asian (CAS); U = undefined; T = T family; and the MANU/ Manu family. We also found some isolates of Mycobacterium bovis. There were no significant association showed between genotype families and gender, but strong significant association found with ethnics and geography. Further confirmation of the results is still ongoing (k value; RFLP and MIRU-VNTR). As conclusion, this study constituted a first attempt to describe the preliminary mapping of genetic population structure of the tubercle bacilli circulating in Indonesia. Key words: preliminary mapping, Mtb complex, spoligotyping, Beijing genotype strain, SpolDB4